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Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Scope The Atlas of Genetics and Cytogenetics in Oncology and Haematology is a peer reviewed on-line journal in open access, devoted to genes, cytogenetics, clinical entities in cancer, and cancer-prone diseases. It presents structured review articles ("cards") on genes, leukaemias, solid tumours, cancer-prone diseases, more traditional review articles on these and also on surrounding topics ("deep insights"), case reports in hematology, and educational items in the various related topics for students in Medicine and in Sciences.

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The Atlas of Genetics and Cytogenetics in Oncology and Haematology (ISSN 1768-3262) is published 6 times a year by ARMGHM, a non profit organisation, and by the INstitute for Scientific and Technical Information of the French National Center for Scientific Research (INIST-CNRS) since 2008. The Atlas is hosted by INIST-CNRS (http://www.inist.fr)

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© ATLAS - ISSN 1768-3262

Atlas Genet Cytogenet Oncol Haematol. 2008;12(3)



Editor-in-Chief

Jean-Loup Huret (Poitiers, France)

Editorial Board Sreeparna Banerjee (Ankara, Turkey) Solid Tumours Section Alessandro Beghini (Milan, Italy) Genes Section Anne von Bergh (Rotterdam, The Netherlands) Genes / Leukaemia Sections Judith Bovée (Leiden, The Netherlands) Solid Tumours Section Vasantha Brito-Babapulle (London, UK) Leukaemia Section Charles Buys (Groningen, The Netherlands) Deep Insights Section Anne Marie Capodano (Marseille, France) Solid Tumours Section Fei Chen (Morgantown, West Virginia) Genes / Deep Insights Sections Antonio Cuneo (Ferrara, Italy) Leukaemia Section Paola Dal Cin (Boston, Massachussetts) Genes / Solid Tumours Section Louis Dallaire (Montreal, Canada) Education Section Brigitte Debuire (Villejuif, France) Deep Insights Section François Desangles (Paris, France) Leukaemia / Solid Tumours Sections Enric Domingo-Villanueva (London, UK) Solid Tumours Section Ayse Erson (Ankara, Turkey) Solid Tumours Section Richard Gatti (Los Angeles, California) Cancer-Prone Diseases / Deep Insights Sections Ad Geurts van Kessel (Nijmegen, The Netherlands) Cancer-Prone Diseases Section Oskar Haas (Vienna, Austria) Genes / Leukaemia Sections Anne Hagemeijer (Leuven, Belgium) Deep Insights Section Nyla Heerema (Colombus, Ohio) Leukaemia Section Jim Heighway (Liverpool, UK) Genes / Deep Insights Sections Sakari Knuutila (Helsinki, Finland) Deep Insights Section Lidia Larizza (Milano, Italy) Solid Tumours Section Lisa Lee-Jones (Newcastle, UK) Solid Tumours Section Edmond Ma (Hong Kong, China) Leukaemia Section Roderick McLeod (Braunschweig, Germany) Deep Insights / Education Sections Cristina Mecucci (Perugia, Italy) Genes / Leukaemia Sections Yasmin Mehraein (Homburg, Germany) Cancer-Prone Diseases Section Fredrik Mertens (Lund, Sweden) Solid Tumours Section Konstantin Miller (Hannover, Germany) Education Section Felix Mitelman (Lund, Sweden) Deep Insights Section Hossain Mossafa (Cergy Pontoise, France) Leukaemia Section Stefan Nagel (Braunschweig, Germany) Deep Insights / Education Sections Florence Pedeutour (Nice, France) Genes / Solid Tumours Sections Elizabeth Petty (Ann Harbor, Michigan) Deep Insights Section Susana Raimondi (Memphis, Tennesse) Genes / Leukaemia Section Mariano Rocchi (Bari, Italy) Genes Section Alain Sarasin (Villejuif, France) Cancer-Prone Diseases Section Albert Schinzel (Schwerzenbach, Switzerland) Education Section Clelia Storlazzi (Bari, Italy) Genes Section Sabine Strehl (Vienna, Austria) Genes / Leukaemia Sections Nancy Uhrhammer (Clermont Ferrand, France) Genes / Cancer-Prone Diseases Sections Dan Van Dyke (Rochester, Minnesota) Education Section Roberta Vanni (Montserrato, Italy) Solid Tumours Section Franck Viguié (Paris, France) Leukaemia Section José Luis Vizmanos (Pamplona, Spain) Leukaemia Section Thomas Wan (Hong Kong, China) Genes / Leukaemia Sections




Volume 12, Number 3, May-June 2008

Table of contents

Gene Section AIFM1 (apoptosis-inducing factor, mitochondrion-associated, 1) 190 Victor J Yuste, Hans K Lorenzo, Santos A Susin

BNIP3 (Bcl-2/adenovirus E1B 19kD-interacting protein 3) 195 Sang-Gi Paik, Hayyoung Lee

BRCA1 (breast cancer 1, early onset) 197 Sreeparna Banerjee

CD97 (CD97 molecule) 201 Gabriela Aust

CDH1 (cadherin 1, type 1, E-cadherin (epithelial)) 204 Marilia de Freitas Calmon, Paula Rahal

GRN (granulin) 208 Hongyong Zhang, Chong-xian Pan, Liang Cheng

HTATIP (HIV-1 Tat interacting protein, 60kDa) 213 Lise Mattera

HYAL1 (hyaluronoglucosaminidase 1) 217 Demitrios H Vynios

MAML2 (mastermind-like 2) 220 Kazumi Suzukawa, Jean-Loup Huret

MUC16 (mucin 16, cell surface associated) 223 Shantibhusan Senapati, Moorthy P Ponnusamy, Ajay P Singh, Maneesh Jain, Surinder K Batra

MUC17 (mucin 17, cell surface associated) 226 Wade M Junker, Nicolas Moniaux, Surinder K Batra

PTHLH (parathyroid hormone-like hormone) 234 Sai-Ching Jim Yeung

SOCS2 (suppressor of cytokine signaling 2) 240 Leandro Fernández-Pérez, Amilcar Flores-Morales

Leukaemia Section del(11)(p12p13) 243 Pieter Van Vlierberghe, Jules PP Meijerin

t(3;5)(q26;q34) 244 Jean-Loup Huret

t(3;9)(q26;p23) 245 Jean-Loup Huret

t(3;17)(q26;q22) 246 Jean-Loup Huret




t(6;7)(q23;q34) 248 Emmanuelle Clappier, Jean Soulier

Solid Tumour Section Soft tissue tumors: Alveolar soft part sarcoma 250 Jean-Loup Huret

Bone: Subungual exostosis with t(X;6)(q13;q22) 253 Clelia Tiziana Storlazzi, Fredrik Mertens

Cancer Prone Disease Section Glomuvenous malformation (GVM) 255 Virginie Aerts, Pascal Brouillard, Laurence M Boon, Miikka Vikkula

Case Report Section Translocation t(1;6)(p35;p25) in B-cell lymphoproliferative disorder with evolution to Diffuse Large B-cell Lymphoma 258 Elvira D Rodrigues Pereira Velloso, Cristina Ratis, Sérgio AB Brasil, João Carlos Guerra, Nydia S Bacal, Cristóvão LP Mangueira

Educational Item Section How human chromosome aberrations are formed 260 Albert Schinzel




Gene Section Review

Atlas Genet Cytogenet Oncol Haematol. 2008;12(3) 190



AIFM1 (apoptosis-inducing factor, mitochondrion-associated, 1) Victor J Yuste, Hans K Lorenzo, Santos A Susin

Cell Death, Senescence and Survival Research Group, Institut de Neurociencies, Universitat Autonoma de Barcelona, Edifici M, Campus de Bellaterra, 08193 Bellaterra Cerdanyola del Valles, Spain (VJY); University of Paris XI, School of Medicine, Hospital Paul Brousse, INSERM U542, 14, av. Paul Vaillant Couturier, 94807 Villejuif, France (HKL); Apoptosis and Immune System, Institut Pasteur, URA 1961-CNRS, 25, rue du Dr. Roux, 75724 Paris Cedex 15, France (SAS)

Published in Atlas Database: October 2007

Online updated version: http://AtlasGeneticsOncology.org/Genes/AIFM1ID44053chXq25.html DOI: 10.4267/2042/38516

This work is licensed under a Creative Commons Attribution-Non-commercial-No Derivative Works 2.0 France Licence. © 2008 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Hugo: AIFM1 Other names: AIF; PDCD8; MGC111425

Location: Xq25 Local order: Centromere (59,500 Kbp)- ARHGEF9 - (…) - RAB33A - AIFM1 - ELF4 - (…) - IL9R -telomere (154,914 Kbp).

AIF gene structure and known isoforms. Genomic organization of AIF and resulting AIF, AIF-exB, AIFsh, AIFsh2, and AIFsh3 mRNA transcripts (schemas in the left). Translation start (ATG, in green) and stop (TGA/TAA, in red) codons are indicated, and the predicted protein product is shown at the right. Numbers in AIF designate exons (in mRNA transcripts) and amino acids (in the predicted proteins). Mitochondria localization signal (MLS), Pyridoxin-redox (Pyr-Redox), nuclear localization sequence (NLS), and C-terminal domains are indicated. I9 (in green) indicates intron 9. The inclusion of the 203-bp exon 9b (lettering in red) produces AIFsh2 and AIFsh3, which encodes 324- and 237-amino acid proteins, respectively. AIFsh2 contains the MLS and the Pyr-Redox domain, but lacks the C-terminal portion of AIF. AIFsh3 has a similar structure as AIFsh2 with the splicing of exon 2, leading to the loss of MLS. Blue lines indicate the splicing of the different isoforms.

AIFM1 (apoptosis-inducing factor, mitochondrion-associated, 1) Yuste VJ, et al.


DNA/RNA Note: AIF (Apoptosis-Inducing Factor). Total gene size 36.471 Kb with a transcribed region of 2.215 Kb which codes for 613 amino acids. To date, five isoforms from AIF gene have been described (AIF, AIFexB, AIFsh, AIFsh2, and AIFsh3).

Description 16 exons spanning 36.471 Kb.

Transcription 2,215 bp mRNA.

Pseudogene Not known.

Figure 1: Schematic model representing the three different AIF forms: precursor, mature, and truncated. AIF is a flavoprotein (with an oxidoreductase enzymatic activity) containing a FAD-bipartite domain (yellow, amino-acids 128-262 and 401-480), a NADH-binding motif (violet, amino-acids 263-400), and a C-terminal domain (red, amino-acids 481-608) where the proapoptotic activity of the protein resides. In addition, it has a Mitochondria Localization Sequence (MLS, in green, amino-acids 1-41) placed in its N-terminal region. Between the first-N-terminal FAD motif and the MLS, AIF possesses a potential Transmembrane Domain (TM, in green, amino-acids 67-83). This TM is flanked by two peptidase-processing positions: a Mitochondrial Processing Peptidase (MPP)-cleavage site (in blue, amino-acid 54) and a calpains- and/or cathepsins-cleavage site (in red, amino-acid 103). Hsp70 (Heat Shock Protein-70) and CypA (Cyclophilin A) bind AIF in amino-acids 150-228 and 367-369, respectively. AIF also possesses two DNA-binding sites, which are located in amino-acids 255-265 and 510-518, respectively. AIF precursor protein has 613 amino-acids. The MPP-mediated cleavage generates the mitochondrial mature AIF (amino-acids 55-613). After an apoptotic insult, calpains or cathepsins cleave AIF to produce truncated-AIF (tAIF), which is released from mitochondria to cytosol (amino-acids 104-613). Figure 2: Ribbon structure of mouse AIF in its mature form (pdb id: 1GV4). As depicted here, three domains are present in the protein. The FAD-binding domain and the NAD-binding domain (yellow) are both similar to oxidoreductase domains from members of the glutathione reductase family. In contrast, the C-terminal domain (blue) displays a particular folding with a specific insertion, which includes residues 580 to 610. This picture also includes the AIF cofactor Flavin Adenine Dinucleotide (FAD; magenta).



Protein Note: 613 amino acids long protein whose structure may be divided into three domains: a FAD-binding domain (residues 128-262 and 401-480), a NADH-binding domain (residues 263-400), and a C-terminal domain (residues 481-608).

Description AIF was initially identified as a protein released from the mitochondrial intermembrane space during the apoptotic process. First studies showed that upon an apoptotic stimulus AIF translocates from mitochondria to cytosol and further to the nucleus where it triggers caspase-independent programmed cell death. AIF, expressed as a precursor of 67 kDa, is addressed to mitochondria by the two MLS placed within the N-terminal prodomain of the protein. Once in mitochondria, this precursor is processed to a mature form of 62 kDa by a first proteolytic cleavage. In this configuration, AIF is an inner-membrane-anchored protein in which the N-terminus is exposed to the mitochondrial matrix and the C-terminal portion to the mitochondrial intermembrane space. AIF is here required for maintenance or maturation of the mitochondrial respiratory chain complex I. After a cell death insult, the 62 kDa AIF-mitochondrial form is cleaved by activated calpains and/or cathepsins to yield a soluble proapoptotic protein with an apparent molecular weight of 57 kDa tAIF (truncated AIF). tAIF is released from mitochondria to cytosol and nucleus to generate two typical hallmarks of caspase-independent

programmed cell death: chromatin condensation and large-scale approximatively 50 kb DNA fragmentation.

Expression Ubiquitously expressed.

Localisation Mitochondrion.

Function AIF has a double life/death function. In its vital role, AIF is required to maintain and/or organize the mitochondrial respiratory complex I, and displays NADH oxidoreductase and peroxide scavenging activities. In addition to this vital function, AIF has been shown to be implicated in programmed cell death (PCD) induction in several experimental models (see bibliography section). In the two most studied AIF-dependent PCD models, AIF death activity is associated with the increase of intracellular Ca2+ (e.g., ischemia/reperfusion injury), or relates with extensive DNA-damage (e.g., treatment with alkylating agents). In the first model, increased intracellular Ca2+ levels trigger depolarization of mitochondrial membrane, subsequent loss of membrane potential, generation of reactive oxygen species (ROS), and AIF mitochondrial release. In the second model, extensive DNA damage, provoked by high doses of alkylating agents such as MNNG or MNU, triggers poly(ADP-ribose) polymerase-1 (PARP-1) over-activation and AIF release from the mitochondrial intermembrane space. This cell death pathway sequentially involves PARP-1, calpains, Bax, and AIF.

Figure 3: Phylogenetic tree representing the relationship between AIF and other oxidoreductases from different species. Note the proximity of the AIF family (red branch) to the NADH-oxidase family from Archaea. The PIR accession codes are enumerated following the abbreviation of each specie: AA: Aquifex aeolicus; AC: Acinetobacter calcoaceticus; AF: Archaeoglobus fulgidus; AT: Arabidopsis thaliana; BC: Burkholderia cepacia; BS: Bacillus subtilis; CE: Caenorhabditis elegans; DD: Dictyostelium discoideum; DM: Drosophila melanogaster; EC: Escherichia coli; HS: Homo sapiens; LS: Lycopersicon esculentum; MJ: Methanocaldococcus jannaschii; MM: Mus musculus; MTH: Methanobacterium thermoautotrophicum; N A: Novosphingobium aromaticivorans; PF: Pseudomonas fluorescens; PH: Pyrococcus horikoshii; PO: Pseudomonas oleovorans; PP: Pseudomonas putida; PS: Pseudomonas sp.; PSA: Pisum sativum; SP: Schizosaccharomyces pombe; SS: Sphingomonas sp.; RE: Rhodococcus erythropolis; RG: Rhodococcus globerulus; XL: Xenopus laevis.



Homology AIF is a highly conserved protein ubiquitously present in all primary kingdoms, Bacteria, Archaea and Eucaryota. The aif gene is inherited from the last universal common ancestor and follows the tree topology with the primary radiation of the archaeo-eukaryotic and bacterial clades. AIF also has a highly significant homology with different families of oxidoreductases, including NADH oxydases, Ascorbate reductases, Glutathione reductases and many NADH-dependent ferredoxin reductases from Archaea and Bacteria to invertebrates and vertebrates. Mouse, Rat homology.

Mutations Note: Several polymorphisms have been identified but none of them has shown any association with a disease.

Implicated in Various cancers Note: Upregulated in cancers (colorectal carcinoma, gastric carcinoma, breast carcinoma and hepatocellular carcinoma, glioblastoma ). AIF expression may play a role in tumor formation and could maintain a transformed state of colon cancer cells involving mitochondrial complex I function.

Cell death Disease AIF has been directly designed as main mediator of cell death in ischemic injuries after overproduction of reactive oxygen species. Indeed, blocking the mitochondrial release of AIF to cytosol and its further nuclear translocation provides protection against neuronal and cardiomyocites cell death. AIF-deficient harlequin mutant mouse presents a significant reduction of neuronal cell death in brain trauma and cerebral ischemia. A similar protective effect was observed in AIF siRNA-treated neurons.

Degenerative disorders Disease AIF is involved in several degenerative disorders. The elevated production of ROS generated in Amyotrophic Lateral Sclerosis, Alzheimer's, or Parkinson diseases concludes in the translocation of AIF. Likewise, AIF release triggered by calpains and cathepsins was observed on in vitro models of Epilepsy and Huntington's disease. AIF-mediated cell death is involved in the pathogenesis of different retinal affections such as retinal detachment, retinitis pigmentosa, or in models of retinal hypoxia. Moreover, an increase of AIF expression has been reported in patients affected with diabetic retinopathy.

References Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE, Brothers GM, Mangion J, Jacotot E, Costantini P, Loeffler M, Larochette N, Goodlett DR, Aebersold R, Siderovski DP, Penninger JM, Kroemer G. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 1999;397(6718):441-446.

Joza N, Susin SA, Daugas E, Stanford WL, Cho SK, Li CY, Sasaki T, Elia AJ, Cheng HY, Ravagnan L, Ferri KF, Zamzami N, Wakeham A, Hakem R, Yoshida H, Kong YY, Mak TW, Zúñiga-Pflücker JC, Kroemer G, Penninger JM. Essential role of the mitochondrial apoptosis-inducing factor in programmed cell death. Nature 2001;410(6828):549-554.

Miramar MD, Costantini P, Ravagnan L, Saraiva LM, Haouzi D, Brothers G, Penninger JM, Peleato ML, Kroemer G, Susin SA. NADH oxidase activity of mitochondrial apoptosis-inducing factor. J Biol Chem 2001;276(19):16391-16398.

Ravagnan L, Gurbuxani S, Susin SA, Maisse C, Daugas E, Zamzami N, Mak T, Jäättelä M, Penninger JM, Garrido C, Kroemer G. Heat-shock protein 70 antagonizes apoptosis-inducing factor. Nat Cell Biol 2001;3(9):839-843.

Klein JA, Longo-Guess CM, Rossmann MP, Seburn KL, Hurd RE, Frankel WN, Bronson RT, Ackerman SL. The harlequin mouse mutation downregulates apoptosis-inducing factor. Nature 2002;419(6905):367-374.

Maté MJ, Ortiz-Lombardía M, Boitel B, Haouz A, Tello D, Susin SA, Penninger J, Kroemer G, Alzari PM. The crystal structure of the mouse apoptosis-inducing factor AIF. Nat Struct Biol 2002;9(6):442-446.

Ye H, Cande C, Stephanou NC, Jiang S, Gurbuxani S, Larochette N, Daugas E, Garrido C, Kroemer G, Wu H. DNA binding is required for the apoptogenic action of apoptosis inducing factor. Nat Struct Biol 2002;9(9):680-684.

Yu SW, Wang H, Poitras MF, Coombs C, Bowers WJ, Federoff HJ, Poirier GG, Dawson TM, Dawson VL. Mediation of poly(ADP-ribose) polymerase-1-dependent cell death by apoptosis-inducing factor. Science 2002;297(5579):259-263.

Bidere N, Lorenzo HK, Carmona S, Laforge M, Harper F, Dumont C, Senik A. Cathepsin D triggers Bax activation, resulting in selective apoptosis-inducing factor (AIF) relocation in T lymphocytes entering the early commitment phase to apoptosis. J Biol Chem 2003;278(33):31401-31411.

Gurbuxani S, Schmitt E, Cande C, Parcellier A, Hammann A, Daugas E, Kouranti I, Spahr C, Pance A, Kroemer G, Garrido C. Heat shock protein 70 binding inhibits the nuclear import of apoptosis-inducing factor. Oncogene 2003;22(43):6669-6678.

Yu SW, Wang H, Dawson TM, Dawson VL. Poly(ADP-ribose) polymerase-1 and apoptosis inducing factor in neurotoxicity. Neurobiol Dis 2003;14(3):303-317. (Review).

Candé C, Vahsen N, Kouranti I, Schmitt E, Daugas E, Spahr C, Luban J, Kroemer RT, Giordanetto F, Garrido C, Penninger JM, Kroemer G. AIF and cyclophilin A cooperate in apoptosis-associated chromatinolysis. Oncogene 2004;23(8):1514-1521.

Gallego MA, Joseph B, Hemström TH, Tamiji S, Mortier L, Kroemer G, Formstecher P, Zhivotovsky B, Marchetti P. Apoptosis-inducing factor determines the chemoresistance of non-small-cell lung carcinomas. Oncogene 2004;23(37):6282-6291.

Vahsen N, Candé C, Brière JJ, Bénit P, Joza N, Larochette N, Mastroberardino PG, Pequignot MO, Casares N, Lazar V, Feraud O, Debili N, Wissing S, Engelhardt S, Madeo F, Piacentini M, Penninger JM, Schägger H, Rustin P, Kroemer G. AIF deficiency compromises oxidative phosphorylation. EMBO J 2004;23(23):4679-4689.



Otera H, Ohsakaya S, Nagaura Z, Ishihara N, Mihara K. Export of mitochondrial AIF in response to proapoptotic stimuli depends on processing at the intermembrane space. EMBO J 2005;24(7):1375-1386.

Polster BM, Basañez G, Etxebarria A, Hardwick JM, Nicholls DG. Calpain I induces cleavage and release of apoptosis-inducing factor from isolated mitochondria. J Biol Chem 2005;280(8):6447-6454.

Urbano A, Lakshmanan U, Choo PH, Kwan JC, Ng PY, Guo K, Dhakshinamoorthy S, Porter A. AIF suppresses chemical stress-induced apoptosis and maintains the transformed state of tumor cells. EMBO J 2005;24(15):2815-2826.

Yuste VJ, Moubarak RS, Delettre C, Bras M, Sancho P, Robert N, d'Alayer J, Susin SA. Cysteine protease inhibition prevents mitochondrial apoptosis-inducing factor (AIF) release. Cell Death Differ 2005;12(11):1445-1448.

Artus C, Maquarre E, Moubarak RS, Delettre C, Jasmin C, Susin SA, Robert-Lézénès J. CD44 ligation induces caspase-independent cell death via a novel calpain/AIF pathway in human erythroleukemia cells. Oncogene 2006;25(42):5741-5751.

Cheung EC, Joza N, Steenaart NA, McClellan KA, Neuspiel M, McNamara S, MacLaurin JG, Rippstein P, Park DS, Shore GC, McBride HM, Penninger JM, Slack RS. Dissociating the dual roles of apoptosis-inducing factor in maintaining mitochondrial structure and apoptosis. EMBO J 2006;25(17):4061-4073.

Delettre C, Yuste VJ, Moubarak RS, Bras M, Lesbordes-Brion JC, Petres S, Bellalou J, Susin SA. AIFsh, a novel apoptosis-inducing factor (AIF) pro-apoptotic isoform with potential pathological relevance in human cancer. J Biol Chem 2006;281(10):6413-6427.

Delettre C, Yuste VJ, Moubarak RS, Bras M, Robert N, Susin SA. Identification and characterization of AIFsh2, a mitochondrial apoptosis-inducing factor (AIF) isoform with NADH oxidase activity. J Biol Chem 2006;281(27):18507-18518.

Jeong EG, Lee JW, Soung YH, Nam SW, Kim SH, Lee JY, Yoo NJ, Lee SH. Immunohistochemical and mutational

analysis of apoptosis-inducing factor (AIF) in colorectal carcinomas. APMIS 2006;114(12):867-873.

Modjtahedi N, Giordanetto F, Madeo F, Kroemer G. Apoptosis-inducing factor: vital and lethal. Trends Cell Biol 2006;16(5):264-272. (Review).

Ruchalski K, Mao H, Li Z, Wang Z, Gillers S, Wang Y, Mosser DD, Gabai V, Schwartz JH, Borkan SC. Distinct hsp70 domains mediate apoptosis-inducing factor release and nuclear accumulation. J Biol Chem 2006;281(12):7873-7880.

Stambolsky P, Weisz L, Shats I, Klein Y, Goldfinger N, Oren M, Rotter V. Regulation of AIF expression by p53. Cell Death Differ 2006;13(12):2140-2149.

Vahsen N, Candé C, Dupaigne P, Giordanetto F, Kroemer RT, Herker E, Scholz S, Modjtahedi N, Madeo F, Le Cam E, Kroemer G. Physical interaction of apoptosis-inducing factor with DNA and RNA. Oncogene 2006;25(12):1763-1774.

Yu SW, Andrabi SA, Wang H, Kim NS, Poirier GG, Dawson TM, Dawson VL. Apoptosis-inducing factor mediates poly(ADP-ribose) (PAR) polymer-induced cell death. Proc Natl Acad Sci USA 2006;103(48):18314-18319.

Boujrad H, Gubkina O, Robert N, Krantic S, Susin SA. AIF-Mediated Programmed Necrosis: A Highly Regulated Way to Die. Cell Cycle 2007 Nov 1;6(21):2612-2619.

Lorenzo HK, Susin SA. Therapeutic potential of AIF-mediated caspase-independent programmed cell death. Drug Resist Updat 2007 December;10(6):235-255

Moubarak RS, Yuste VJ, Artus C, Bouharrour A, Greer PA, Menissier-de Murcia J, Susin SA. Sequential activation of poly(ADP-ribose) polymerase 1, calpains, and Bax is essential in apoptosis-inducing factor-mediated programmed necrosis. Mol Cell Biol 2007;27(13):4844-4862.

This article should be referenced as such:

Yuste VJ, Lorenzo HK, Susin SA. AIFM1 (apoptosis-inducing factor, mitochondrion-associated, 1). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):190-194.

Gene Section Mini Review




BNIP3 (Bcl-2/adenovirus E1B 19kD-interacting protein 3) Sang-Gi Paik, Hayyoung Lee

Department of Biology, School of Biosciences and Biotechnology, Chungnam National University, Daejeon 305-764, Korea


Online updated version: http://AtlasGeneticsOncology.org/Genes/BNIP3ID822ch10q26.html DOI: 10.4267/2042/38517


Identity Hugo: BNIP3 Other names: NIP3 Location: 10q26.3

DNA/RNA

Description 14.23 kb on reverse strand; 6 exons

Transcription mRNA in MCF-7 cells are 1.7kb (major) and 1.5 kb (minor) and 1.3 kb (minor).

Protein

Domain map of BNIP3 protein; BH3 domain (Bcl-2 holomogy 3

domain); TM domain (transmembrane domain)

Description 194 amino acids; 1 BH3 domain and 1 TM domain; BH3 only Bcl2 family member. The TM domain and C-terminal tail are essential for mitochondrial membrane localization and proapoptotic function. The predicted molecular weight is 21.5 kDa. BNIP3 migrates as 30 kDa monomeric form and 60 kDa dimeric form on SDS-PAGE.

Expression BNIP3 is detected in mouse oviduct, uterus, spleen, lung, stomach, brain, seminal, lacrimal, submaxillary, heart, kidney, liver. It can be detected in cell lines such as HeLa, 293T, RAW264.7 and K562 cells. Its expression can be induced in both normal and cancer tissues that experience hypoxia or hypoxia-like conditions. Other stimuli, such as nitric oxide or arsenic trioxide, are also reported to induce BNIP3 expression.

Localisation Outer mitochondrial membrane.

Function Proapoptotic protein; BNIP3 leads to opening of the mitochondrial permeability transition pore (PTP) thereby abolishing the proton electrochemical gradient and this is followed by chromatin condensation and DNA fragmentation. BNIP3 leads necrosis-like apoptosis. Unusually to the other Bcl-2 family proteins, the BNIP3-induced cell death depends not on BH3 domain but on C-terminal TM domain. BNIP3-induced cell death is known to be independent the nuclear translocation of AIF. However, whether caspase activation and cytochrome c release are involved in the cell death remains controversial. BNIP3 can induce autophagy. However whether the consequence of the autophagy is the cell death or survival remains to be established. Since BNIP3 is induced by hypoxia through transcription factor HIF-1, it was postulated to play a role in hypoxia-induced cell death. Hypoxia-induced acidosis augments the proapoptotic function of BNIP3.

Homology The close homologue: BNIP3L/BNIP3a/Nix/B5 (8q21).

BNIP3 (Bcl-2/adenovirus E1B 19kD-interacting protein 3) Paik SG, Lee H


The BH3-only Bcl2 family members: BBC3/PUMA (19q13), BCL2L11/BIM/BOD (2q13), BID (22q11), BIK/NBK/BBC1 (22q13), BLK (8q23), BMF (15q14), HRK/DP5/BID3 (12q24), PMAIP1/NOXA (18q21).

Implicated in Pancreatic cancer Prognosis Pancreatic adenocarcinoma is highly resistant to chemical and radiation therapy, and has an extremely poor prognosis. Reduced expression of BNIP3 increased resistance to gemcitabine and 5-fluoro-uracil (5-FU) and showed a good correlation with reduced patient survival.

Oncogenesis In most cases of pancreatic adenocarcinoma, BNIP3 expression was not detected even in response to hypoxia. The promoter of BNIP3 is located within a CpG island and is methylated in most pancreatic cancer cell lines. Restoration of BNIP3 expression by the methyltransferase inhibitor, 5-aza-deoxycytidine, induced death of pancreatic cancer cells in response to hypoxia.

Colorectal cancer Oncogenesis Methylation of BNIP3 in 66% of primary colorectal cancer.

References Boyd JM, Malstrom S, Subramanian T, Venkatesh LK, Schaeper U, Elangovan B, D'Sa-Eipper C, Chinnadurai G. Adenovirus E1B 19 kDa and Bcl-2 proteins interact with a common set of cellular proteins. Cell 1994;79:341-351.

Chen G, Ray R, Dubik D, Shi L, Cizeau J, Bleackley RC, Saxena S, Gietz RD, Greenberg AH. The E1B 19K/Bcl-2-binding protein Nip3 is a dimeric mitochondrial protein that activates apoptosis. J Exp Med 1997;186:1975-1983.

Yasuda M, Theodorakis P, Subramanian T, Chinnadurai G. Adenovirus E1B-19K/BCL-2 interacting protein BNIP3 contains a BH3 domain and a mitochondrial targeting sequence. J Biol Chem 1998;273:12415-12421.

Bruick RK. Expression of the gene encoding the proapoptotic Nip3 protein is induced by hypoxia. Proc Natl Acad Sci USA 2000;97:9082-9087.

Ray R, Chen G, Vande Velde C, Cizeau J, Park JH, Reed JC, Gietz RD, Greenberg AH. BNIP3 heterodimerizes with Bcl-2/Bcl-X(L) and induces cell death independent of a Bcl-2 homology 3 (BH3) domain at both mitochondrial and nonmitochondrial sites. J Biol Chem 2000;275:1439-1448.

Vande Velde C, Cizeau J, Dubik D, Alimonti J, Brown T, Israels S, Hakem R, Greenberg AH. BNIP3 and genetic control of

necrosis-like cell death through the mitochondrial permeability transition pore. Mol Cell Biol 2000;20:5454-5468.

Kim JY, Cho JJ, Ha J, Park JH. The carboxy terminal C-tail of BNip3 is crucial in induction of mitochondrial permeability transition in isolated mitochondria. Arch Biochem Biophys 2002;398:147-152.

Kubasiak LA, Hernandez OM, Bishopric NH, Webster KA. Hypoxia and acidosis activate cardiac myocyte death through the Bcl-2 family protein BNIP3. Proc Natl Acad Sci USA 2002;99:12825-12830.

Okami J, Simeone DM, Logsdon CD. Silencing of the hypoxia-inducible cell death protein BNIP3 in pancreatic cancer. Cancer Res 2004;64:5338-5346.

Yook YH, Kang KH, Maeng O, Kim TR, Lee JO, Kang KI, Kim YS, Paik SG, Lee H. Nitric oxide induces BNIP3 expression that causes cell death in macrophages. Biochem Biophys Res Commun 2004;321:298-305.

Abe T, Toyota M, Suzuki H, Murai M, Akino K, Ueno, M, Nojima M, Yawata A, Miyakawa H, Suga T, Ito H, Endo T, Tokino T, Hinoda Y, Imai K. Upregulation of BNIP3 by 5-aza-2'-deoxycytidine sensitizes pancreatic cancer cells to hypoxia-mediated cell death. J Gastroenterol 2005;40:504-510.

Akada M, Crnogorac-Jurcevic T, Lattimore S, Mahon P, Lopes R, Sunamura M, Matsuno S, Lemoine NR. Intrinsic chemoresistance to gemcitabine is associated with decreased expression of BNIP3 in pancreatic cancer. Clin Cancer Res 2005;11:3094-3101.

Erkan M, Kleeff J, Esposito I, Giese T, Ketterer K, Buchler MW, Giese NA, Friess H. Loss of BNIP3 expression is a late event in pancreatic cancer contributing to chemoresistance and worsened prognosis. Oncogene 2005;24:4421-4432.

Murai M, Toyota M, Satoh A, Suzuki H, Akino K, Mita H, Sasaki Y, Ishida T, Shen L, Garcia-Manero G, Issa JP, Hinoda Y, Tokino T, Imai K. Aberrant DNA methylation associated with silencing BNIP3 gene expression in haematopoietic tumours. Br J Cancer 2005;92:1165-1172.

Murai M, Toyota M, Suzuki H, Satoh A, Sasaki Y, Akino K, Ueno M, Takahashi F, Kusano M, Mita H, Yanagihara K, Endo T, Hinoda Y, Tokino T, Imai K. Aberrant methylation and silencing of the BNIP3 gene in colorectal and gastric cancer. Clin Cancer Res 2005;11:1021-1027.

Webster KA, Graham RM, Bishopric NH. BNip3 and signal-specific programmed death in the heart. J Mol Cell Cardiol 2005;38:35-45.

An HJ, Maeng O, Kang KH, Lee JO, Kim YS, Paik SG, Lee H. Activation of Ras upregulates pro-apoptotic BNIP3 in nitric oxide-induced cell death. J Biol Chem 2006;281:33939-33948.

Lee H, Paik SG. Regulation of BNIP3 in normal and cancer cells. Mol Cells 2006;21:1-6. (Review).

Bacon AL, Fox S, Turley H, Harris AL. Selective silencing of the hypoxia-inducible factor 1 target gene BNIP3 by histone deacetylation and methylation in colorectal cancer. Oncogene 2007;26:132-141.


Paik SG, Lee H. BNIP3 (Bcl-2/adenovirus E1B 19kD-interacting protein 3). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):195-196.

Gene Section Review




BRCA1 (breast cancer 1, early onset) Sreeparna Banerjee

Department of Biology, Middle East Technical University, Ankara 06531, Turkey


Online updated version: http://AtlasGeneticsOncology.org/Genes/BRCA1ID163ch17q21.html DOI: 10.4267/2042/38518


Identity Hugo: BRCA1 Other names: BRCAI; BRCC1; IRIS; PSCP; RNF53 Location: 17q21.31 Local order: According to NCBI Map Viewer, genes flanking BRCA1 in centromere to telomere direction on 17q21 are: VAT1 17q21 (vesicle amine transport protein 1 homolog (T californica)); RND2 17q21 Rho family GTPase 2; RPL21P4 17q21 ribosomal protein L21 pseudogene 4; BRCA1 17q21 breast cancer 1, early onset; NBR2 17q21 neighbour of BRCA1 gene; BRCA1P1 17q21 BRCA1 pseudogene 1; NBR1 17q21.31 neighbour of BRCA1 gene. Note: BRCA1 is a tumour suppressor phosphoprotein that combines with other tumour suppressors, DNA damage and repair proteins, and signal transducers to form a large multi-subunit protein complex known as BRCA1-associated genome surveillance complex (BASC). Truncating mutations and missence mutations in the BRCA1 gene are found in a large number of familial breast cancer cases. Individuals who inherit a germline mutation of BRCA1 or BRCA2 have a significantly increased lifetime risk for the development of breast and/or ovarian cancer.

DNA/RNA Note: The subcellular localization and physiological function of this gene is greatly modulated by the several alternately splices isoforms that are found. Several of these alternatively spliced transcript variants have been described, however, not all have had their full-length natures identified.

Description According to Entrez-Gene, BRCA1 gene maps to NC_000017.9 in the region between 38449840 and 38530994 on the minus strand and spans across 81.1 kilo bases. According to Spidey (mRNA to genomic sequence alignment tool, http://www.ncbi.nlm.nih.gov/spidey), BRCA1 has 24 exons, the sizes being 181, 99, 54, 78, 89, 140, 105, 47, 77, 89, 172, 127, 191, 311, 88, 78, 41, 84, 55, 74, 61, 1506.

Transcription BRCA1 mRNA NM_007302.3 has 7388 bps. The BRCA1 gene contains two separate promoters that induce transcription of mRNAs with different 5'UTRs, a shorter 5'UTRa and a longer 5'UTRb. The downregulation of BRCA1 gene expression in certain breast cancers is caused by a switch from expression of a 5'UTRa, which enables efficient translation, to expression of 5'UTRb, which contains secondary structure and upstream open reading frames that strongly inhibit translation.

Pseudogene According to Entrez Gene the BRCA1 pseudogene 1 (BRCA1P1) is located on 17q21.

Protein Note: BRCA1 sequence is not well conserved between mammals, however, two domains, the C terminal BRCT (BRCA1 C Terminal) motifs and the N-terminal RING domain are highly conserved.

BRCA1 (breast cancer 1, early onset) Banerjee S


The BRCA1 protein showing the RING finger domain, the Nuclear Localisation Signal domain and the BRCT domains. AA- amino acids.

Description BRCA1 is an 1863 amino acid 220kDa protein with an E3 ubiquitin ligase activity as well as a phospho-peptide binding activity. It has several domains that are essential for its function as depicted in the figure. The RING finger domain of BRCA1, commonly found in many DNA repair proteins, consists of a conserved core of approximately 50 amino acids in a pattern of seven cysteine residues and one histidine residue to form a structure that can bind to two Zn++ ions. This motif aids in mediating protein-protein interaction, as exemplified by the interaction of BRCA1 with BARD1 (BRCA1 associated RING domain). This interaction is critical since mutations in the Zn++ binding regions, crucial for heterodimerization with BARD1, have been found in tumours. BRCA1 accumulates in distinct foci in the nucleus during S phase and this transfer is aided by its Nuclear Localisation Signal (NLS) domain. A further role of BARD1 is also implicated whereby its association with the RING finger domain of BRCA1 is necessary for the transfer of BRCA1 to the nucleus. BRCA1 interacts with Rad50 of the MRN complex through the region AA 341-748 and can directly bind to branched, flap and four way DNA structures through a central domain spanning residues 452-1079. The protein inhibits the nucleolytic activities of the Mre11/Rad50/Nbs1 complex as a result of this direct DNA binding. The C terminus of BRCA1, which can function as a transcriptional activation domain, consists of two tandemly arranged elements called BRCT (BRCA1 C- terminal). This motif specifically binds to phosphorylated proteins, an event that is commonly associated with DNA damage response. BRCA1 is capable of interacting directly with BRCA2 and with Rad51 via BRCA2 through this motif. Another protein that interacts with BRCA1 via BRCT is the BRCA1 associated C-terminal helicase (BACH1). BACH1 is said to aid BRCA1 in the DNA damage response and maintain the protein at the nuclear foci formed after DNA damage response. Other proteins that can interact with BRCA1 through the BRCT domains are C terminal Interacting protein /CtIP), RNA Polymerase II, BACH 1 (a member of DEAH helicase family) and p53.

Expression BRCA1 is ubiquitously expressed in humans with the highest levels observed in the ovaries, testis and thymus. It is a tumour suppressor and a reduced expression is correlated with the transformation procedure and aetiology of sporadic breast cancer. This reduction is expression is said to be transcriptionally regulated with implications of aberrant promoter methylation at CpG dinucleotides as well as CREB binding sites.

Localisation Located in the nucleus.

Function Role of BRCA1 in DNA repair: BRCA1 is a part of a large complex of proteins, the BASC, which monitors the genome for damage and signals downstream effectors. BRCA1 has been implicated in two pathways of DNA double strand break repair: homologous recombination (HR) and non homologous end joining (NHEJ). Upon exposure to DNA damaging agents, BRCA1 becomes hyperphosphorylated and is rapidly relocated, along with Rad51, to sites of DNA synthesis marked by proliferating cell nuclear antigen (PCNA). Rad51, a homolog of the bacterial RecA, is a central player in HR, catalyzing the invasion of the single stranded DNA in a homologous duplex and facilitating the homology search during the establishment of joint molecules. A recent study, however, has indicated that BRCA1 deficient breast cancer cells compensate for this deficiency by upregulating Rad51. The resultant HR may be erroneous and thereby lead to tumorigenesis. In addition, BRCA1 is said to inhibit the MRN complex which is is implicated in bringing together two DNA strands together for the error prone NHEJ. BRCA1-deficient cells are sensitive to ionizing radiation and DNA damaging drugs, such as mitomycin C. Transcriptional regulation: BRCA1 is capable of transcriptional regulation and chromatin remodelling when tethered to promoters of genes important in the DNA repair process and breast cancer markers. It is a



member of the core RNA polymerase II transcriptional machinery, a feature exploited by the DNA damage recognition process. In addition, BRCA1 interacts with p300/CBP, transcriptional coactivators for CREB. p300/CBP are inhibited by the viral oncoprotein E1A and the functionality of E1A as an oncogene could be in part caused by an obstruction of BRCA1:p300/CBP cooperation resulting in the loss of the tumour-suppressing function of BRCA1. BRCA1 can act as a transcriptional coactivator or co repressor of proteins implicated in chromatin remodelling, such as the histone deacetylase complexes or components of the SWI/SNF-related chromatin-remodelling complex. Cell Cycle Regulation by BRCA1: BRCA1, based on its phosphorylation status, elicits DNA damage induced cell cycle arrest at several stages through modulation of specific downstream target genes. BRCA1 transactivates p21cip1/WAF1, which contributes to an arrest at the G1/S boundary. ATM phosphorylation of BRCA1 appears to be important for its role in the intra S phase checkpoint activation. BRCA1 is also implicated in the transcriptional regulation of several genes such as cyclinB, 14-3-3sigma, GADD45, wee-1 kinase and PLK1 associated with the G2/M checkpoint. p53-dependent apoptosis: The BRCA1 protein is capable of physically interacting with the p53 tumour suppressor gene, and can stimulate p53-dependent transcription from the p21WAF1/CIP1 mdm2 and promoters. In addition, the BRCA1-BARD1 complex is required for the phosphorylation of p53 at Ser15 by ATM/ATR following DNA damage by IR or UV radiation. The phosphorylation of p53 at Ser-15 is essential for the G(1)/S cell cycle arrest via transcriptional induction of the cyclin-dependent kinase inhibitor p21 after DNA damage. Ubiquitination: BRCA1 and BARD1 interact together to form an E3 ubiquitin ligase. RNA polII stalled at sites of DNA damage is a target for this ubiquitin ligase mediated degradation following DNA damage, thereby allowing access to the repair machinery. BRCA1 ubiquitinates the transcriptional preinitiation complex, not for proteasomal degradation, but to prevent a stable association of TFIIE and TFIIH; thereby blocking the initiation of mRNA synthesis.

Homology Dog (Canis familiaris): BRCA1; Chimpanzee (Pan troglodytes): BRCA1; Rat (Rattus norvegicus): Brca1; Mouse (Mus musculus): Brca1; Chicken (Gallus gallus): BRCA1.

Mutations Note: BRCA1 germline mutations contribute significantly to the development of familial/hereditary breast and ovarian cancer. However, each gene carries as many as 1000 different disease associated mutations,

many of which are rare. These mutations are distributed uniformly along the entire coding region and intronic sequences flanking each exon. The mutations are at a high penetrance therefore women who carry these mutations have a lifetime risk of 80-90% to develop breast cancer. Founder mutations such as the BRCA1-185delAG and 5382insC are found among Ashkenazi Jews. Larger and complex genomic rearrangements in the exons 21 and 22 of the BRCA1 gene, resulting in a lack of the BRCT motif have been reported.

Implicated in Breast cancer Disease Heterozygous carriers of high-risk mutations in the general Caucasian population have been estimated to be about one in 1000 for the BRCA1 gene. The lifetime risk of the development of hereditary breast cancer with the presence of BRCA1 mutations is very high. In addition, for sporadic breast cancer, a reduction in the expression of BRCA1 rather than the presence of mutations has been observed. The lack of a functional BRCA1 leads to impaired repair of DNA double strand breaks, cell cycle progression and transcriptional regulation, thereby causing the development of neoplasms.

Ovarian cancer Disease Mutations of the BRCA1 gene is the major cause for familial breast and ovarian cancer incidence. The lifetime risks of ovarian cancer associated with a BRCA1 gene mutation carrier has been estimated as 40 to 50%. The most common mutations are frameshift and nonsense mutations that are predicted to cause premature truncation of the BRCA1 protein. In addition, mutations that are predicted to affect splice-site consensus sequences as well as missense mutation have also been seen in ovarian cancer. Large genomic alterations, such as the gains in copy number of exon 13 as well as deletion of exons in the BRCA1 gene is also associated with the development of ovarian cancer.

Other cancers Disease An increased relative risk to the development of cancer of the colon, cervix, uterus, pancreas and prostate has been suggested in BRCA1-mutation carriers.

References Miki Y, Swensen J, Shattuck-Eidens D, Futreal PA, Harshman K, Tavtigian S, Liu Q, Cochran C, Bennett LM, Ding W, et al. A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science 1994;266(5182):66-71.

Baumann P, Benson FE, West SC. Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro. Cell 1996;87(4):757-766.



Chapman MS, Verma IM. Transcriptional activation by BRCA1. Nature 1996;382(6593):678-679.

Ruffner H, Verma IM. BRCA1 is a cell cycle-regulated nuclear phosphoprotein. Proc Natl Acad Sci USA 1997;94(14):7138-7143.

Scully R, Anderson SF, Chao DM, Wei W, Ye L, Young RA, Livingston DM, Parvin JD. BRCA1 is a component of the RNA polymerase II holoenzyme. Proc Natl Acad Sci USA 1997;94(11):5605-5610.

Scully R, Chen J, Ochs RL, Keegan K, Hoekstra M, Feunteun J, Livingston DM. Dynamic changes of BRCA1 subnuclear location and phosphorylation state are initiated by DNA damage. Cell 1997;90(3):425-435.

Chen J, Silver DP, Walpita D, Cantor SB, Gazdar AF, Tomlinson G, Couch FJ, Weber BL, Ashley T, Livingston DM, Scully R. Stable interaction between the products of the BRCA1 and BRCA2 tumor suppressor genes in mitotic and meiotic cells. Mol Cell 1998;2(3):317-328.

Ouchi T, Monteiro AN, August A, Aaronson SA, Hanafusa H. BRCA1 regulates p53-dependent gene expression. Proc Natl Acad Sci USA 1998;95(5):2302-2306.

Rice JC, Massey-Brown KS, Futscher BW. Aberrant methylation of the BRCA1 CpG island promoter is associated with decreased BRCA1 mRNA in sporadic breast cancer cells. Oncogene 1998;17(14):1807-1812.

Chai YL, Cui J, Shao N, Shyam E, Reddy P, Rao VN. The second BRCT domain of BRCA1 proteins interacts with p53 and stimulates transcription from the p21WAF1/CIP1 promoter. Oncogene 1999;18(1):263-268.

Zhong Q, Chen CF, Li S, Chen Y, Wang CC, Xiao J, Chen PL, Sharp ZD, Lee WH. Association of BRCA1 with the hRad50-hMre11-p95 complex and the DNA damage response. Science 1999;285(5428):747-750.

Pao GM, Janknecht R, Ruffner H, Hunter T, Verma IM. CBP/p300 interact with and function as transcriptional coactivators of BRCA1. Proc Natl Acad Sci USA 2000;97(3):1020-1025.

Wang Y, Cortez D, Yazdi P, Neff N, Elledge SJ, Qin J. BASC, a super complex of BRCA1 associated proteins involved in the recognition and repair of aberrant DNA structures. Genes Dev 2000;14:927-939.

Atlas E, Stramwasser M, Mueller CR. A CREB site in the BRCA1 proximal promoter acts as a constitutive transcriptional element. Oncogene 2001;20(48):7110-7114.

Hashizume R, Fukuda M, Maeda I, Nishikawa H, Oyake D, Yabuki Y, Ogata H, Ohta T. The RING heterodimer BRCA1-BARD1 is a ubiquitin ligase inactivated by a breast cancer-derived mutation. J Biol Chem 2001;276(18):14537-14540.

Paull TT, Cortez D, Bowers B, Elledge SJ, Gellert M. Direct DNA binding by BRCA1. Proc Natl Acad Sci USA 2001;98:6086-6091.

Fabbro M, Rodriguez JA, Baer R, Henderson BR. BARD1 induces BRCA1 intranuclear foci formation by increasing RING-dependent BRCA1 nuclear import and inhibiting BRCA1 nuclear export. J Biol Chem 2002;277(24):21315-21324.

Jasin M. Homologous repair of DNA damage and tumorigenesis: the BRCA connection. Oncogene 2002;21(58):8981-8993.

Sobczak K, Krzyzosiak WJ. Structural determinants of BRCA1 translational regulation. J Biol Chem 2002;277(19):17349-17358.

Fabbro M, Savage K, Hobson K, Deans AJ, Powell SN, McArthur GA, Khanna KK. BRCA1-BARD1 complexes are required for p53Ser-15 phosphorylation and a G1/S arrest following ionizing radiation-induced DNA damage. J Biol Chem 2004;279(30):31251-31258.

Scully R, Xie A, Nagaraju G. Molecular functions of BRCA1 in the DNA damage response. Cancer Biol Ther 2004;3(6):521-527.

Durant ST, Nickoloff JA. Good timing in the cell cycle for precise DNA repair by BRCA1. Cell Cycle 2005;4(9):1216-1222.

Boulton SJ. Cellular functions of the BRCA tumour suppressor proteins. Biochem Soc Trans 2006;34(Pt 5):633-645.

Cantor SB, Andreassen PR. Assessing the link between BACH1 and BRCA1 in the FA pathway. Cell Cycle 2006;5(2):164-167.

Mullan PB, Quinn JE, Harkin DP. The role of BRCA1 in transcriptional regulation and cell cycle control. Oncogene 2006;25(43):5854-5863.

Peng M, Litman R, Jin Z, Fong G, Cantor SB. BACH1 is a DNA repair protein supporting BRCA1 damage response. Oncogene 2006;25(15):2245-2253.

Walsh T, Casadei S, Coats KH, Swisher E, Stray SM, Higgins J, Roach KC, Mandell J, Lee MK, Ciernikova S, Foretova L, Soucek P, King MC. Spectrum of mutations in BRCA1, BRCA2, CHEK2, and TP53 in families at high risk of breast cancer. JAMA 2006;295(12):1379-1388.

Ferla R, Calo V, Cascio S, Rinaldi G, Badalamenti G, Carreca I, Surmacz E, Colucci G, Bazan V, Russo A. Founder mutations in BRCA1 and BRCA2 genes. Ann Oncol 2007;18 Suppl 6:vi93-98.

Horwitz AA, Affar el B, Heine GF, Shi Y, Parvin JD. A mechanism for transcriptional repression dependent on the BRCA1 E3 ubiquitin ligase. Proc Natl Acad Sci USA 2007;104(16):6614-6619.

Martin RW, Orelli BJ, Yamazoe M, Minn AJ, Takeda S, Bishop DK. RAD51 Up-regulation Bypasses BRCA1 Function and Is a Common Feature of BRCA1-Deficient Breast Tumors. Cancer Research 2007;67:9658-9665.

Oldenburg RA, Meijers-Heijboer H, Cornelisse CJ, Devilee P. Genetic susceptibility for breast cancer: how many more genes to be found?. Crit Rev Oncol Hematol 2007;63(2):125-149.

Ramus SJ, Harrington PA, Pye C, Dicioccio RA, Cox MJ, Garlinghouse-Jones K, Oakley-Girvan I, Jacobs IJ, Hardy RM, Whittemore AS, Ponder BA, Piver MS, Pharoah PD, Gayther SA. Contribution of BRCA1 and BRCA2 mutations to inherited ovarian cancer. Hum Mutat 2007;28(12):1207-1215.

Zikan M, Pohlreich P, Stribrna J, Kleibl Z, Cibula D. Novel complex genomic rearrangement of the BRCA1 gene. Mutat Res 2007; 637 (1-2):205-208.


Banerjee S. BRCA1 (breast cancer 1, early onset). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):197-200.

Gene Section Review




CD97 (CD97 molecule) Gabriela Aust

University of Leipzig, Faculty of Medicine, Research Laboratories, Center of Surgery, Liebigstr. 20, Leipzig, D-04103, Germany Published in Atlas Database: October 2007

Online updated version: http://AtlasGeneticsOncology.org/Genes/CD97ID996ch19p13.html DOI: 10.4267/2042/38519


Identity Hugo: CD97 Other names: TM7LN1 Location: 19p13

DNA/RNA Description DNA contains 27.322 kb composed of 20 coding exons. Exons 1-2 encode the 5' untranslated region and the signal peptide, exons 3-7 the five EGF domains, exons 8-13 the extracellular stalk, exons 14-18 the seven-span transmembrane (TM7) domains and exons 19-20 the intracellular part and the 3' untranslated region.

Transcription 3247 bp mRNA transcribed in telomeric to centromeric

orientation; 2508 bp open reading frame. Human CD97 exists in three isoforms that result from alternative splicing of exons 5 and 6 and thus contain different numbers of EGF domains in the extracellular part of the molecule. The isoforms are designated as CD97 (EGF1,2,5), CD97 (EGF1,2,3,5) and CD97 (EGF1-5) in human.

Pseudogene No pseudogenes reported.

Protein Description CD97 belongs to the B family of G protein-coupled receptors (GCPRs). Subfamily B2 contains cell surface molecules with long extracellular N-termini (LNB-TM7) known also as adhesion class of heptahelical receptors.

Genomic organization of CD97 (drawn to scale), boxes represent exons.

Structure of CD97. Three isoforms containing 3, 4, or 5 EGF domains exist. N-glycosylation sites in the EGF domains are indicated.

CD97 (CD97 molecule) Aust G


CD97 is the founding member of a small subfamily within the adhesion class called EGF-TM7 family. All EGF-TM7 receptors (CD97, EMR1, EMR2, EMR3, EMR4) consist of extracellular tandemly arranged EGF domains, a stalk, the seven-span transmembrane (TM7) und a short intracellular part. They are expressed as heterodimers of a non-covalently bound alpha- and beta-chain resulting from intracellular autocatalytic cleavage at a conserved GCPR proteolytic site (GPS). The alpha-chain represents the extracellular region with the varying numbers of EGF domains and the main part of the stalk and the beta-chain consists of the stalk residue, the TM7 and intracellular part. Three CD97 isoforms containing 3, 4 or 5 EGF domains are described. The mature full length proteins contain either 722, 766 or 815 amino acids (aa). After cleavage the (secretory) alpha-chains contain 420, 464, or 513 aa. The beta-chain theoretically contains 305 aa with a molecular weight of 34.3 kDa. However, immunoprecipitation of the beta-chain yielded a molecular weight of approximately 28 kDa. The discrepancy between the theoretical and actual molecular weight of the beta-chain is not yet clarified. Depending on the cell type and transformation status of the cell, CD97 is completely or partly N-glycosylated or naked. In normal muscle cells CD97 is not or only slightly N-glycosylated. The molecular weight for the respective naked alpha-chain of the various CD97 isoforms are 45.6, 50.5 and 55.8 kDa. In hematopoetic cells CD97 is N-glycosylated at the EGF domains resulting in molecular weights of 74-78, 80-82, and 86-89 kDa for the alpha-chains of the respective isoform. During tumor transformation CD97 may get N-glycosylated. Although the CD97 stalk contains many Ser or Thr residues the molecule seems not to be O-glycosylated.

Expression Broad, not cell-type specific. - Hematopoetic system: strong in peripheral blood myeloid cells and activated lymphocytes, moderately in subsets of tissue-derived leukocytes; - Strong in smooth muscle cells (except for arterial vascular smooth muscle cells), skeletal muscle cells (stronger in slow-twitch fibers), heart muscle cells; - Fat cells; - Low in normal intestinal, thyroidal epithelial cells, moderately in duct cells of the pancreas, parotis gland and in bile duct cells of the liver.

Localisation Usually at the cell membrane; soluble CD97 (sCD97) representing the CD97 alpha-chain in body fluids; Skeletal muscle cells: at the sarcolemm and intracellularly in the sacroendoplasmatic reticulum (SR).

Function CD97 has the ability to bind cellular and extracellular matrix ligands. The first two EGF domains of CD97 bind CD55 (decay accelerating factor). The fourth EGF domain of CD97 and thus only the longest CD97 isoform interacts with the glycosaminoglycan chondroitin sulfate B. CD97 binds to alpha5beta1 and alphavbeta3 integrins through interaction with the CD97 stalk region. - Hematopoetic cells: Functional studies indicate a role of CD97 in leukocyte trafficking. CD97 antibodies block tissue localization of immune cells in vivo leading to impaired protection against bacteria and amelioration of autoimmune pathology. - Tumor cells: In vitro CD97 increases single cell random motility and directed migration and invasion of tumor cells in 2D and 3D matrices. CD97 enhances proteolytic activity of matrix metalloproteinases (MMPs) and secretion of chemokines in an isoform-specific manner. CD97 (EGF 1,2,5) overexpression promotes tumor growth in scid mice. The alpha-chain of the longest CD97 (EGF1-5) isoform (sCD97) enhances angiogenesis in in vivo tumor models. - Muscle, fat, duct cells: function unknown.

Homology H. sapiens: CD97 P. troglodytes: CD97 B. taurus: CD97 S. scrofa: CD97 C. lupus: CD97 M. musculus: CD97 R. norvegicus: CD97 Exists only in mammals.

Mutations Note: unknown.

Implicated in Note: Note for all tumors:

Antibodies to various epitopes of CD97 vary strongly in their staining pattern and cross-reactivity to other EGF-TM7 molecules. The first group of monoclonal antibodies, which includes BL-Ac/F2, VIM-3b and CLB-CD97/1, binds to the EGF domains of CD97 (CD97EGF/ antibodies). These antibodies also detect EMR2, another member of the EGF-TM7 family. In most cases, this cross-reactivity will not influence the results obtained for CD97 staining in tumors since EMR2 is strongly restricted to myeloid cells. CD97 antibodies MEM-180 and CLB-CD97/3 bind to the stalk region of CD97 (CD97stalk) and do not bind EMR2.

CD97 (CD97 molecule) Aust G


CD97EGF epitope accessibility depends on cell type-specific N-glycosylation (see above). CD97EGF antibodies detect only N-glycosylated CD97. During tumor transformation, not only the CD97 protein expression level but also the degree of CD97 N-glycosylation varies. Thus, the selection of the CD97 antibody strongly influences the result in immunohistological studies focused on the correlation between CD97 and histopathological subtypes, diagnosis, progression, or prognosis of tumors. CD97 in tumors is strongly regulated at the post-trancriptional level.

Thyroid cancer Note: In normal thyroid tissue, no or low immunoreactivity of CD97 is found. In differentiated follicular thyroid carcinoma or papillary thyroid carcinoma, CD97 expression is also either lacking or low. Most undifferentiated anaplastic carcinomas reveal high CD97 presentation. CD97 is absent or only weakly present in patients with postoperative T1 tumors but increased greatly with the progression to postoperative T4 tumors. Until now, only antibodies against CD97 EGF domains (CD97EGF antibodies, see above) have been used in studies of thyroid carcinomas.

Prognosis Not determined.

Cytogenetics Not determined.

Oncogenesis Overexpression of CD97 might be important for the progression of thyroid cancer.

Colorectal cancer Note: Normal human colorectal epithelium is slightly CD97-positive. Most colorectal carcinomas express CD97. The strongest staining for CD97 occurs in scattered tumor cells at the invasion front compared to cells located within solid tumor formations of the same tumor. Carcinomas with more strongly CD97-stained scattered tumor cells show a poorer clinical stage as well as increased lymph vessel invasion compared to cases with uniform CD97 staining.



Oncogenesis Overexpression of CD97 might be important for invasion and metastasis of colorectal cancer. Gastric cancer Note: CD97 is present in normal parietal cells of

gastric mucosa. It is stronger expressed by most gastric carcinomas. Half of the tumors show scattered tumor cells at the invasion front with stronger CD97 expression than tumor cells located in solid tumor formations.



Leiomyosarcoma Note: Normal smooth muscle cells are CD97-positive. In this cell type CD97 is not N-glycosylated. Thus, monoclonal antibodies that detect an N-glycosylation dependent epitop of CD97 do not react with normal smooth muscle cells (CD97EGF antibodies). During transformation CD97 get partly N-glyocosylated in most uterine leiomyoma and or completely N-glyocosylated in nearly 25% of the leiomyosarcomas. These tumors are now positive for CD97EGF antibodies. However, one third of leiomyosarcomas are completely devoid of CD97.



References Aust G, Eichler W, Laue S, Lehmann I, Heldin NE, Lotz O, Scherbaum WA, Dralle H, Hoang-Vu C. CD97: A dedifferentiation marker in human thyroid carcinomas. Cancer Res 1997;57:1798-1806.

Aust G, Steinert M, Schütz A, Wahlbuhl M, Hamann J, Wobus M. CD97, but not its closely related EGF-TM7 family member EMR2, is expressed on gastric, pancreatic and esophageal carcinomas. Am J Clin Pathol 2002;118:699-707.

Steinert M, Wobus M, Boltze C, Schütz A, Wahlbuhl M, Hamann J, Aust G. Expression and regulation of CD97 in colorectal carcinoma cell lines and tumor tissues. Am J Pathol 2002;161:1657-1667.

Wang T, Ward Y, Tian L, Lake R, Guedez L, Stetler-Stevenson WG, Kelly K. CD97, an adhesion receptor on inflammatory cells, stimulates angiogenesis through binding integrin counter receptors on endothelial cells. Blood 2004;105:2836-2844.

Aust G, Wandel E, Boltze C, Sittig D, Schutz A, Horn LC, Wobus M. Diversity of CD97 in smooth muscle cells (SMCs). Cell Tissue Res 2006;323:1-9.

Galle J, Sittig D, Hanisch I, Wobus M, Wandel E, Loeffler M, Aust G. Individual cell - based models of tumor - environment interactions. Multiple effects of CD97 on tumor invasion. Am J Pathol 2006;169:1802-1811.


Aust G. CD97 (CD97 molecule). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):201-203.

Gene Section Review




CDH1 (cadherin 1, type 1, E-cadherin (epithelial)) Marilia de Freitas Calmon, Paula Rahal

Laboratory of Genomics studies, São Paulo State University, Department of Biology, São José do Rio Preto - SP, Brasil


Online updated version: http://AtlasGeneticsOncology.org/Genes/CDH1ID166ch16q22.html DOI: 10.4267/2042/38520


Identity Hugo: CDH1 Other names: Arc-1; CD324; CDHE; Cadherin-1; E-cadherin; ECAD; LCAM; UVO; Uvomorulin Location: 16q22.1

DNA/RNA Description DNA contains 98250 bp composed of 16 coding exons.

Transcription 4828 bp mRNA transcribed in centromeric to telomeric orientation; 2649 bp open reading frame.

Pseudogene Yes, for example, the repeat sequence named c41-cad is a pseudogene of the cadherin family. c41-cad is localizated on 5q13.

DNA of CDH1 gene composed of 16 coding exons.

Protein Description The cadherins are a family of calcium-dependent transmembrane linker proteins; the first three that were discovered were named according to their tissue origin (E-cadherin from epithelium, N-cadherin from neural tissue and P-cadherin from placenta). The mature E-cadherin protein consists of three major domains: a large extracellular portion (exons 4-13), which mediates homophilic cellular interactions; and smaller transmembrane (exons 13-14) and cytoplasmic domains (exons 14-16), the latter providing a link to the

actin cytoskeleton through an association with various catenins, such as B-catenin. The protein E-cadherin is a calcium-dependent cell-cell adhesion molecule expressed in adherents junctions between epithelial cells. It is a transmembrane glycoprotein with five extracellular domains that mediate intercellular adhesion through homophilic binding. The cytoplasmatic domain is bound to the actin cytoskeleton via intracellular attachment proteins, the catenins. The actin cytoskeleton forms a transcellular network mediating structural integrity, cellular polarity and epithelial morphogenesis.

Expression Present tissue specificity for non-neural epithelial tissues and there are high levels in solid tissues.

Localisation Cell junction; single-pass type I membrane protein. Anchored to actin microfilaments through association with alpha-catenin, beta-catenin and gamma-catenin. Sequential proteolysis induced by apoptosis or calcium influx, results in translocation from sites of cell-cell contact to the cytoplasm.

Function One of the most important and ubiquitous types of adhesive interactions required for the maintenance of solid tissues is that mediated by the classic cadherin adhesion molecules. Cadherins are transmembrane Ca2+- dependent homophilic adhesion receptors that are well known to play important roles in cell recognition and cell sorting during development. However, they continue to be expressed at high levels in virtually all solid tissues. There are many members of the classic cadherin family (which is a subset of the larger cadherin superfamily), but E-cadherin in epithelial tissues has been the most studied in the context of stable adhesions.

CDH1 (cadherin 1, type 1, E-cadherin (epithelial)) Calmon MF, Rahal P


Three-dimensional structure of the beta-catenin arm repeat region in complex with the E-cadherin cytoplasmic domain (Huber and Weis, 2001). The arm repeats are formed by three helices, H1 and H2 (both gray) and H3 (blue). Residues 134-161, which include part of the alpha-catenin-binding site and a portion of the first arm repeat, form a single helix in this particular crystal structure (cyan). E-cadherin is divided into five regions of primary structure (1-5) that are indicated in distinct colors (Pokutta S and Weis WI, 2007).

Continued expression and functional activity of E-cadherin are required for cells to remain tightly associated in the epithelium, and in its absence the many other cell adhesion and cell junction proteins expressed in epithelial cells (see below) are not capable of supporting intercellular adhesion. In its capacity to maintain the overall state of adhesion between epithelial cells, E-cadherin is thought to act as an important suppressor of epithelial tumor cell invasiveness and metastasis.

Homology Pan troglodytes - CDH1; Canis lupus familiaris - CDH1; Mus musculus - Cdh1; Rattus norvegicus - Cdh1; Gallus gallus - LOC415860; Danio rerio - cdh1.

Mutations Germinal 30 CDH1 germline mutations have been described in hereditary diffuse gastric cancer families. 25 have been inactivating (frameshift, nonsense, and splice-site), the remainders are missense. The mutations are distributed equally throughout the gene.

Somatic Somatically acquired mutations in CDH1 were found in about 56% of lobular breast tumors, generally (>90%) in combination with loss of the wild-type allele, while no mutations were found in ductal primary breast carcinomas. Most of these somatic mutations result in premature stop codons as a consequence of insertions, deletions and nonsense mutations. As the majority of these frameshift and nonsense mutations is predicted to

generate secreted E-cadherin fragments, the functionality of this major cell-cell adhesion protein is lost. Other cancer-confined E-cadherin mutations also result in crippled proteins. The distinctive invasive growth pattern, which is typical for lobular breast cancers, is fully compatible with this functional inactivation. 472 human tumors and 15 different cancer cell lines derived from 10 different tissues have been screened for CDH1 mutation. So far, frequent somatic mutations (50%) have been identified only in sporadic diffuse gastric cancer (DGC), Lobular Breast Cancer. For sporadic DGC, most somatic mutations are missense (exons 8, 9) or exon skipping. For sporadic Lobular Breast Cancer, most somatic mutations are truncating.472 human tumors and 15 different cancer cell lines derived from 10 different tissues have been screened for CDH1 mutation. So far, frequent somatic mutations (50%) have been identified only in sporadic Diffuse Gastric Cancer, Lobular Breast Cancer. Interestingly, there is a major difference between the mutation types identified in these two carcinoma types. In diffuse gastric carcinomas, the predominant mutations are exon skippings causing in-frame deletions. By contrast, most mutations identified in lobular breast cancer result in premature stop codons. In the case of the diffuse gastric carcinomas, a mutation cluster region is suggested as more than 60% of mutations cause exon skipping of exon 8 and 9. Preliminary in vitro studies using transfected cell lines suggest that tumor-associated E-cadherin mutations reduce cell adhesion, increase cell motility, and change cell morphology possibly by dominant negative mechanisms.



57 CDH1 mutations have been found to date. 50 of these are listed in Human Gene Mutation Database. Truncating (27) and splice site (7) mutations are found above the Schema (34/45, 76%), missense mutations below it (11/45, 24%). Two marked with an asterisk have been reported as somatic mutations in sporadic diffuse gastric cancer. No Polymorphisms. No Gross deletions/duplications, complex rearrangements, repeat variations been reported. They spread out all over CDH1 gene (Brooks-Wilson et al., 2004).

On the contrary, the truncating mutations present in lobular breast cancers are obviously scattered over the entire E-cadherin gene. In line with this finding is the observation that the expression of E-cadherin protein is lost in lobular breast cancers, in contrast to the retention of expression of the mutant E-cadherin proteins in diffuse gastric carcinomas. Surprisingly, so far almost no E-cadherin mutations have been found to be located in the highly conserved cytoplasmic domain. In most cases, E-cadherin mutations are found in combination with loss of the wild-type allele.

Implicated in Non-small cell lung cancer Prognosis Reduced E-cadherin correlates with lymph node metastasis. The rate of vascular invasion was statistically high in cases with the reduced expression of E-cadherin. Reduction of E-cadherin is associated with the degree of differentiation. Bohm et al. found a correlation between differentiation and E-cadherin expression in lung squamous cell carcinoma, and Bongiorno et al. found that well-differentiated lung cancers express E-cadherin, in a preserved fashion, and that poorly differentiated tumors exhibited a reduced or disorganized staining pattern. Sulzer et al. also found that E-cadherin expression significantly correlated with increasing tumor differentiation. In general, undifferentiated or poorly differentiated cancer cells tend to have a strong potential to invade tissues. These results suggest that reduction of E-cadherin correlates with tumor invasion.

Oncogenesis Reduced E-cadherin expression weakens cell-to-cell attachment, and tumor cells detach from the primary tumor, invade vessels, and migrate to lymph nodes. Once tumor cells reattach to lymph nodes, E-cadherin is strongly expressed, and lymph nodes are subject to metastases.

Melanoma Oncogenesis The major adhesion mediator between keratinocytes and normal melanocytes is E-cadherin, which disappears during melanoma progression. While normal melanocytes express E-cadherin, this molecule is not found on nevus or melanoma cells. The loss of E-cadherin likely plays a crucial role in tumor progression. Cells that have lost epithelial differentiation, as manifested by the loss of functional E-cadherin, show increased mobility and invasiveness. Keratinocytes can no longer control melanoma cells that have lost E-cadherin. When melanoma cells are forced to express E-cadherin and are cocultured with keratinocytes, they dramatically change: melanomas adhere to keratinocytes, no longer express invasion-related molecules, and lose their invasive capacities

Oesophageal adenocarcinoma Prognosis Reduction in the expression of E-cadherin in patients with OSCC was shown to be strongly associated with postoperative blood borne recurrence, resulting in a poorer prognosis than in those patients with tumours showing normal expression before surgery.



This finding suggested that in patients with reduced E-cadherin immunoreactivity, the metastatic potential of the oesophageal cancer cells may be increased. Therefore, the evaluation of E-cadherin immunoreactivity may be useful in predicting haematogenous spread and hence recurrence, thus serving as an aid for planning adjuvant treatment after surgery in patients with OSCC. It has also been reported that E-cadherin might be an independent predictor of micrometastasis in lymph nodes that are classified as N0 by routine histopathological analysis.

References Bohm M, Totzeck B, Birchmeier W, Wieland I. Differences of E-cadherin expression levels and patterns in primary and metastatic human lung cancer. Clin. Exp. Metastasis 1994;12:55-62.

Berx G, Cleton-Jansen A-M, Nollet F, de Leeuw WJF, van de Vijver MJ, Cornelisse C, van Roy F. E-cadherin is a tumor/invasion suppressor gene mutated in human lobular breast cancers. EMBO J 1995;14(24):6107-6115.

Berx G, Staes K, van Hengel J, Molemans F, Bussemakers MJG, van Bokhoven A, van Roy F. Cloning and characterization of the human invasion suppressor gene E-cadherin (CDH1). Genomics 1995;26:281-289.

Bongiorno PF, al-Kasspooles M, Lee SW, Rachwal WJ, Moore JH, Whyte RI, Orringer MB, Beer DG. E-cadherin expression in primary and metastatic thoracic neoplasms and in Barrett’s oesophagus. Br. J. Cancer 1995;71:166-172.

Selig S, Bruno S, Scharf JM, Wang CH, Vitale E, Gilliam TC, Kunkel LM. Expressed cadherin pseudogenes are localized to the critical region of the spinal muscular atrophy gene. Proc. Natl Acad Sci USA 1995;92:3702-3706.

Berx G, Cleton-Jansen A-M, Strumane K, de Leeuw WJF, Nollet F, van Roy FM, Cornelisse C. E-cadherin is inactivated in a majority of invasive human lobular breast cancers by truncation mutations throughout its extracellular domain. Oncogene 1996;13:1919-1925.

Gumbiner BM. Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell 1996;84:345-357. (Review)

Berx G, Nollet F, Strumane K, van Roy F. An efficient and reliable multiplex PCR/SSCP mutation analysis test applied to the human E-cadherin gene. Hum Mutat 1997;9(6):567-574.

Berx G, Becker KF, Höfler H, Van Roy F. Mutations of the Human E-Cadherin (CDH1) gene. Human mutation 1998;12:226-237.

Sulzer MA, Leers M P, Van N JA, Bollen EC, Theunissen PH. Reduced E-cadherin expression is associated with increased lymph node metastasis and unfavorable prognosis in non-small cell lung cancer. Am J Resp Crit Care 1998;157(41):1319-1323.

Kase S, Sugio K, Yamazaki K, Okamoto T, Yano T, Sugimachi K. Expression of E-cadherin and ß-Catenin in Human Non-Small Cell Lung Cancer and the Clinical Significance. Clinical Cancer Research 2000;6:4789-4796.

Berx G, Van Roy F. The E-cadherin/catenin complex: an important gatekeeper in breast cancer tumorigenesis and malignant progression. Breast Cancer Res 2001;3(5):289-293.

Cleton-Jansen A. E-cadherin and loss of heterozygosity at chromosome 16 in breast carcinogenesis: different genetic pathways in ductal and lobular breast cancer? Breast Cancer Research 2002;4:5-8.

Brooks-Wilson AR, Kaurah P, Suriano G, Leach S, Senz J, Grehan N, Butterfield YSN, Jeyes J, Schinas J, Bacani J, Kelsey M, Ferreira P, MacGillivray B, MacLeod P, Micek M, Ford J, Foulkes W, Australie K, Greenberg C, LaPointe M, Gilpin C, Nikkel S, Gilchrist D, Hughes R, Jackson CE, Monaghan KG, Oliveira MJ, Seruca R, Gallinger S, Caldas C; Huntsman D. Germline E-cadherin mutations in hereditary diffuse gastric cancer: assessment of 42 new families and review of genetic screening criteria. Journal of Medical Genetics 2004;41:508-517.

Perlis C, Herlyn M. Recent Advances in Melanoma Biology. The Oncologist 2004;9(2):182-187.

Sweet KM, Lynch HT. Genetic aetiology of diffuse gastric cancer: so near, yet so far. Journal of Medical Genetics 2004;41:481-484.

Nair KS, Naidoo R, Chetty R. Expression of cell adhesion molecules in oesophageal carcinoma and its prognostic value. Journal of Clinical Pathology 2005;58(4):343-351.

Pokutta S, Weis WL. Structure and mechanism of cadherins and catenins in cell-cell contacts. Annu Rev Cell Dev Biol. 2007;23:237-61.


Calmon MF, Rahal P. CDH1 (cadherin 1, type 1, E-cadherin (epithelial)). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):204-207.

Gene Section Review




GRN (granulin) Hongyong Zhang, Chong-xian Pan, Liang Cheng

UC Davis Cancer Center, 2700 Stockton Blvd, Oak Park Research Building, Ste 2301, Univ. of California at Davis, Sacramento, CA 95817, USA (HZ); Division of Hematology/Oncology, Univ. of California at Davis, 4501 X Street, Rm 3016, Sacramento, CA 95817, USA (CXP); Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, Clarian Pathology Laboratory Room 4010, 350 West 11th Street, Indianapolis, IN 46202, USA (LC)


Online updated version: http://AtlasGeneticsOncology.org/Genes/GRNID40757ch17q21.html DOI: 10.4267/2042/38521


Identity Hugo: GRN Other names: GEP; GP88; PCDGF; PEPI; PGRN; acrogranin; granulin-epithelin; proepithelin; progranulin Location: 17q21.32

DNA/RNA Description 13 exons, including 12 protein encoding exons and a further 5' non-coding exon.

Transcription Major mRNA: 2323bp

Protein Description Granulins are a family of secreted, glycosylated peptides; Granulins are cleaved from a single precursor protein with 7.5 repeats of a highly conserved 12-cysteine granulin/epithelin motif. The 88 kDa precursor protein, progranulin, is also called proepithelin and PC cell-derived growth factor. Cleavage of the signal peptide produces mature granulin which can be further cleaved into a variety of active, 6 kDa peptides. These smaller cleavage products are named granulin A, granulin B, and granulin C, etc.

Expression Granulins are widely expressed. Normally, high levels of GRN expression on rapidly proliferating cells, such as skin cells, deep crypts of gastrointestinal tract,

kidney, and immune cells; Low levels of GRN expression on muscle and liver cells. However, over-expressing on some kinds of tumor cells, such as breast cancer, prostate cancer, ovarian cancer.

Localisation Nucleus.

Function Progranulin stimulates cell proliferation, migration and survival. It activates conventional growth factor signaling pathways including the p44/42 MAPkinase and phosphatidylinositol 3-kinase pathways and the Focal Adhesion Kinase pathway. Many experiments show that increasing the expression of progranulin can stimulate the tumor growth on immortalized but otherwise non-tumorigenic cells. SW13 cells overexpress progranulin (high PGRN), so, they produce large tumors in nude mice; cells that express less progranulin (basal PGRN), do not grow as tumors. However, progranulin is necessary for tumor growth. Attenuating progranulin (PCDGF) expression in mammary cancer cells MDA-MB-468 and human hepatocellular carcinoma cell lines (HepB3) leaded to a dramatic reduction (90% and 87%, respectively) in the size of tumors when the cells were grown in nude mice. Also, some experiments indicated that progranulin caused an increase of the motility and the invasiveness of tumor and played an important effect on apoptosis of tumor cells, reduced the rate of cell death.

Mutations Note: Mutations in the progranulin (PGRN) gene have been identified in frontotemporal lobar degeneration with ubiquitin inclusions linked to chromosome 17q21.

GRN (granulin) Zhang H, et al.


There are two novel frameshift mutations and three possible pathogenic missense mutations in the PGRN gene, and PGRN mutations in familial cases recruited from a large population-based study of frontotemporal lobar degeneration carried out in the Netherlands. However, no mutation was found in the development of different cancers.

Implicated in Breast cancer Disease Progranulin has been shown to play a major role in breast tumorigenesis by stimulating proliferation, mediating survival and conferring resistance to some chemicals such as tamoxifen and doxorubicin, and its overexpression account for the resistance to therapeutic agents. PCDGF/GP88 has metastatic potential in breast cancer, and tumor cells (such as MCF-7 cells) with a PCDGF over-expression or treated exogenously with PCDGF both stimulated anchorage-independent cell growth and accelerated cell migration through matrigel. Furthermore, PCDGF/GP88 also can up-regulated the expression of matrix metalloprotease-9, and stimulated VEGF expression in some tumor cells. So, PCDGF/GP88 could act to promote metastasis and angiogenesis in human breast cancer cells in addition to stimulating their proliferation and survival. PCDGF/GP88 activated mitogen-activated protein kinase (MAP kinase Erk1/Erk2) as well as phosphatidylinositol 30-kinase (PI-3 kinase) pathways leading to the stimulation of several cyclins including Cyclin D1 and Cyclin B. In the adrenal carcinoma SW-13 cells, progranulin expression was also a major determinant of focal adhesion kinase signaling pathway in addition to MAP kinase and PI-3 kinase.

Prognosis GRN might play an important role in deciding the behavior of node-positive breast cancer, so, GRN maybe provide valuable information for the prognosis of breast cancer patients. Since all the in vitro studies indicated the importance of PCDGF/GP88 in breast tumorigenesis. PCDGF/GP88 expression was then examined in pathological samples. Correlation studies between PCDGF expression and prognostic markers such as ER/PR expression, proliferation index Ki67, p53, and erbB2 were also conducted. Normally, PCDGF staining was observed in breast carcinoma, whereas it was not detected in benign breast epithelium. In breast carcinoma, PCDGF expression was more common in ductal carcinoma than in invasive lobular carcinoma. Moreover, PCDGF staining was almost never observed in lobular carcinoma in situ, whereas most of ductal carcinoma in situ (DCIS) expressed PCDGF. PCDGF expression in DCIS correlated strongly with nuclear grade in DCIS and histological grades in IDC. Both ER positive and ER

negative tumors had moderate to strong PCDGF expression. Positive correlation was found between PCDGF staining and Ki67 proliferation index. Similarly, a larger percentage of tumors with moderate/strong PCDGF expression were p53 positive. In contrast, PCDGF expression was independent of cerbB-2 overexpression. This study provides evidence of the high incidence of PCDGF expression in human breast cancer with positive correlation with clinicopathological variables such as tumor grade, proliferation index, and p53 expression. These characteristics the absence of expression in benign breast tissue suggest an important role of PCDGF in breast cancer pathogenesis and make it a potential novel target for the treatment of breast cancer.

Prostate Cancer Disease Normal prostate tissue did not express, or expressed low levels of PCDGF. PCDGF expression could be detected in more than 50% of cells in all specimens of prostatic intraepithelial neoplasia (PIN) and invasive prostate cancer. The expression of PCDGF in normal prostate tissue was much less intense and in a smaller fraction of cells than in PIN and invasive adenocarcinoma (P less than 0.0001). There was no correlation of PCDGF expression with age, Gleason score, pathological stage, status of lymph node metastasis, extraprostatic extension, perineural invasion, surgical margins, and vascular invasion. So, the induction of PCDGF expression occurs during the development of PIN. PCDGF may be a new molecular target for the treatment and prevention of prostate cancer.

Ovarian Carcinoma Disease The GEP/PCDGF has been shown to be an important growth and survival factor induced by low-malignant-potential (LPA) and ET-1 and cAMP/EPAC through ERK1/2 for ovarian cancer cells, and its expression is a predictor of patient survival in metastatic ovarian cancer cells. The prosurvival function of GEP is important in ovarian cancer tumor progression and chemoresponse. Overexpression of GEP increased capacity to migrate and invade their substratum, and was associated with cisplatin chemoresistance. Meantime, GEP overexpression increased tumor formation and protected cells from tumor regression in response to cisplatin treatment in vivo.

Prognosis Several experiments discovered and validated the differential expression of GEP between noninvasive LPA tumors and invasive epithelial ovarian cancers in an effort to define a molecular basis for the pathologic differences between these epithelial tumor subtypes. Low malignant potential tumors share cytologic



similarities with invasive ovarian cancers but have epithelial cells that lack the capacity to invade their underlying stroma. These tumors are slow growing and rarely metastasize and patients with LMP tumors present most often with disease limited to the ovary. This presentation translates into a marked improved clinical outcome over patients with invasive ovarian cancers, with over 95% of patients alive at 10 years. In contrast, patients with invasive ovarian cancers more commonly present with, and die of, disseminated disease and have a 40% overall 5-year survival. GEP expression also was observed in primary and metastatic epithelial ovarian carcinoma specimens, with down-regulated expression in tumor cells of malignant effusions. The poor outcome associated with stromal GEP expression suggests a prognostic role for this growth factor in ovarian carcinoma.

Endometrial cancer Disease The majority of endometrial cancers arise as a result of estrogen stimulation, the molecular targets of which remain incompletely defined. GEP may be one such target. GEP co-expression with ER was observed in most of cancers examined. A two to fivefold increase in GEP expression with estradiol and/or tamoxifen treatment was observed in KLE cells. Silencing of GEP in HEC-1-A cells using shRNA resulted in a decrease in proliferation among transfected cells. However, co-expression of GEP and ER in endometrial cancer cells, and the regulation of GEP by estrogen, suggests a role for GEP in steroid-mediated endometrial cancer cell growth. Further, characterization of GEP as a steroid-mediated growth factor in these cells may help me to understand endometrial cancer biology very well.

Teratoma Disease The PC cell line is a highly tumorigenic, insulin-independent, teratoma-derived cell line isolated from the nontumorigenic, insulin-dependent 1246 cell line. Studies of the PC cell growth properties have led to the purification of an 88-kDa secreted glycoprotein called PC cell-derived growth factor (PCDGF), which has been shown to stimulate the growth of PC cells as well as 3T3 fibroblasts. Since PCDGF was isolated from highly tumorigenic cells, its level of expression was examined in PC cells as well as in nontumorigenic and moderately tumorigenic cells from which PC cells were derived, and the levels of PCDGF mRNA and protein were very low in the nontumorigenic cells and increased in tumorigenic cell lines in a positive correlation with their tumorigenic properties. An inhibition of PCDGF expression resulted in a dramatic inhibition of tumorigenicity of the transfected cells when compared with empty-vector control cells. These data demonstrate the importance in tumor formation of overexpression of the novel growth factor PCDGF.

Brain tumor-glioblastoma multiforme Disease The 2.1-kb granulin mRNA was expressed predominantly in glial tumors, whereas expression was not detected in non-tumor brain tissues. Granulin may be a glial mitogen, as addition of synthetic granulin peptide to primary rat astrocytes and three different early-passage human glioblastoma cultures increased cell proliferation in vitro, whereas increasing concentrations of granulin antibody inhibited cell growth in a dose-dependent manner. The differential expression pattern, tissue distribution, and implication of this glioma-associated molecule in growth regulation suggest a potentially important role for granulin in the pathogenesis and/or malignant progression of primary brain neoplasms.

Multiple Myeloma Disease PCDGF mRNA and protein expression was detected in human MM cell lines such as ARP-1 and RPMI 8226, and PCDGF added exogenously stimulated cell growth and sustained cell survival of both ARP-1 and RPMI 8226 cells in a dose- and time-dependent fashion. When treated with neutralizing anti-PCDGF antibody, RPMI 8225 cells growth was inhibited. This indicated that PCDGF acts as an autocrine growth factor for MM cells. Studies of signal transduction pathways showed PCDGF stimulated mitogen-activated protein kinase and phosphatidylinositol 3'-kinase pathways but not the Janus-activated kinase-signal transducer and activator of transcription pathway. Immunohistochemical analysis of bone marrow smears obtained from MM patients indicated that PCDGF expression was associated with myeloma cells from MM patients and correlated with the presence of MM disease.

Laryngeal carcinoma Disease

The PC cell-derived growth factor protein levels and mRNA levels of the laryngeal squamous cell carcinomas were significantly higher than those of normal laryngeal tissues. Simultaneously, the difference in the levels of mRNA and protein between those of laryngeal precancerous lesions (papilloma/leukoplakia) and those of normal tissues was significant, whereas those of laryngeal precancerous lesions (papilloma/leukoplakia) were significantly lower than those of laryngeal squamous cell carcinomas. Strong PC cell-derived growth factor expression was associated with lymph node metastases in laryngeal squamous cell carcinoma. Functional studies on Hep-2 cell lines demonstrated that the attenuation of PC cell-derived growth factor expression levels led to diminished cell proliferation rates, anchorage-independent growth in vitro, tumor forming in vivo and resistance to apoptosis. PC cell-derived



growth factor is a pivotal autocrine growth factor in the tumorigenesis of laryngeal squamous cell carcinoma. In the future, PC cell-derived growth factor may be a logical and potential target for early diagnosis, specific therapy and prognosis of laryngeal squamous cell carcinoma.

Bladder Cancer Disease Proepithelin is overexpressed in bladder cancer cell lines and clinical specimens of bladder cancer. Proepithelin did not appreciably affect cell growth, but it did promote migration of 5637 bladder cancer cells and stimulate in vitro wound closure and invasion. These effects required the activation of the mitogen-activated protein kinase pathway and paxillin, which upon proepithelin stimulation formed a complex with focal adhesion kinase and active extracellular signal-regulated kinase. However, proepithelin plays a role in stimulating migration, invasion of bladder cancer cells, and establishing of the invasive phenotype.

Prognosis So far, it is unclear if proepithelin has an significant effect on the prognosis of patient with bladder cancer.

Renal epithelium Disease Acrogranin levels were low in benign renal tissue and increased in malignant renal tissue. In addition, high-grade RCC exhibited higher levels of expression than low-grade RCC and normal tissue. So, acrogranin may be a functional important growth factor in RCC and a potential molecular marker for high-grade RCC.

Hepatocellular carcinoma Disease In hepatocellular carcinoma, there is a closed relationship between p53 and GEP protein. Studies revealed an overall positive association between the two protein expression patterns, and the association of p53 and GEP protein expression was found to be highly significant only in HCCs with wild-type p53; there was no association in HCCs with p53 mutation. The GEP levels in the HepG2 hepatoma cell line with a wild-type p53 background were modulated by transfection experiments. Overexpression of the GEP protein resulted in an increased p53 protein level and suppression of the GEP protein resulted in a decreased p53 protein level in HepG2 cells. In summary, p53 wild-type protein nuclei accumulation is associated with GEP protein expression in human HCC specimens, and GEP modulates p53 wild-type protein levels in vitro.

Gastric cancer Disease Granulin was expressed in gastric cancer cells, and may be considered as a tumor associated antigen.

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Wang W, Hayashi J, Kim WE, Serrero G. PC cell-derived growth factor (granulin precursor) expression and action in human multiple myeloma. Clin Cancer Res 2003;9(6):2221-2228.

Boudreau CR, Liau LM. Molecular characterization of brain tumors. Clin Neurosurg 2004;51:81-90.

Cheung ST, Wong SY, Leung KL, Chen X, So S, Ng IO, Fan ST. Granulin-epithelin precursor overexpression promotes growth and invasion of hepatocellular carcinoma. Clin Cancer Res 2004;10(22):7629-7636.

Davidson B, Alejandro E, Flørenes VA, Goderstad JM, Risberg B, Kristensen GB, Trope CG, Kohn EC. Granulin-epithelin precursor is a novel prognostic marker in epithelial ovarian carcinoma. Cancer 2004;100(10):2139-2147.

Pan CX, Kinch MS, Kiener PA, Langermann S, Serrero G, Sun L, Corvera J, Sweeney CJ, Li L, Zhang S, Baldridge LA, Jones TD, Koch MO, Ulbright TM, Eble JN, Cheng L. PC cell-derived growth factor expression in prostatic intraepithelial neoplasia and prostatic adenocarcinoma. Clin Cancer Res 2004;10(4):1333-1337.

Tangkeangsirisin W, Serrero G. PC cell-derived growth factor (PCDGF/GP88, progranulin) stimulates migration, invasiveness and VEGF expression in breast cancer cells. Carcinogenesis 2004;25(9):1587-1592.

Kamrava M, Simpkins F, Alejandro E, Michener C, Meltzer E, Kohn EC. Lysophosphatidic acid and endothelin-induced proliferation of ovarian cancer cell lines is mitigated by neutralization of granulin-epithelin precursor (GEP), a prosurvival factor for ovarian cancer. Oncogene 2005;24(47):7084-7093.

Asaka S, Fujimoto T, Akaishi J, Ogawa K, Onda M. Genetic prognostic index influences patient outcome for node-positive breast cancer. Surg Today 2006;36(9):793-801.

Cheung ST, Wong SY, Lee YT, Fan ST. GEP associates with wild-type p53 in hepatocellular carcinoma. Oncol Rep 2006;15(6):1507-1511.

Jones MB, Houwink AP, Freeman BK, Greenwood TM, Lafky JM, Lingle WL, Berchuck A, Maxwell GL, Podratz KC, Maihle NJ. The granulin-epithelin precursor is a steroid-regulated growth factor in endometrial cancer. J Soc Gynecol Investig 2006;13(4):304-311.

Monami G, Gonzalez EM, Hellman M, Gomella LG, Baffa R, Iozzo RV, Morrione A. Proepithelin promotes migration and invasion of 5637 bladder cancer cells through the activation of ERK1/2 and the formation of a paxillin/FAK/ERK complex. Cancer Res 2006;66(14):7103-7110.

Wang W, Hayashi J, Serrero G. PC cell-derived growth factor confers resistance to dexamethasone and promotes tumorigenesis in human multiple myeloma. Clin Cancer Res 2006;12(1):49-56.

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Kong WJ, Zhang SL, Chen X, Zhang S, Wang YJ, Zhang D, Sun Y. PC cell-derived growth factor overexpression promotes proliferation and survival of laryngeal carcinoma. Anticancer Drugs 2007;18(1):29-40.

Miyanishi M, Mandai M, Matsumura N, Yamaguchi K, Hamanishi J, Higuchi T, Takakura K, Fujii S. Immortalized ovarian surface epithelial cells acquire tumorigenicity by Acrogranin gene overexpression. Oncol Rep 2007;17(2):329-333.

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Zhang H, Pan CX, Cheng L. GRN (granulin). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):208-212.

Gene Section Review




HTATIP (HIV-1 Tat interacting protein, 60kDa) Lise Mattera

Dr Trouche Team, LBCMCP, UMR 5088 CNRS, 118 route de Narbonne, 31062 Toulouse cedex 9, France


Online updated version: http://AtlasGeneticsOncology.org/Genes/HTATIPID40893ch11q13.html DOI: 10.4267/2042/38522


Identity Hugo: HTATIP Other names: Tip60; Tip; 60kDa Tat interacting protein; HIV-1 Tat interacting protein; cPLA(2) interacting protein; iTip60; PLIP/Tip60b; Tip60a; Esa1; Hs.6364 Location: 11q13.1

DNA/RNA

Description The HTATIP gene consists of 14 exons. 7,586 bases.

Transcription The predominant mRNA transcribed from this gene is 2,229 bp long. This is actually the isoform 2 of HTATIP. Two others isoforms generated by alternative splicing have been described: - Isoform 1 retains the alternatively spliced intron 1, - Isoform 3 lacks exon 5.

Pseudogene No pseudogene is currently known.

Protein Description The Tip60 protein (isoform 2) is 513 amino acids long and its molecular weight is about 60 kDa. It was cloned and characterized in 1996 thanks to its interaction with the HIV-1 transactivator Tat protein. Isoform 1 produces a 546 amino acids long protein. Isoform 3 produces a 461 amino acids long protein. A novel isoform, Tip55, encodes a novel splicing variant corresponding to 103 amino acids of the C-terminus. The domain architectures of human TIP60 is similar to yeast Esa1 protein and consist of a chromodomain and a MYST domain harboring a zinc finger and an Acetyl-CoA binding site.

Expression Tip60 is ubiquitously expressed. In mouse adult tissues Tip60 is expressed in the following decreasing order of intensity: testis, heart, brain, kidney, liver, lung, with little to no expression in spleen and skeletal muscle. In human, Tip60 (Isoform 2) and PLIP (Isoform 3) are expressed in human heart, kidney and brain tissue. With a half-life of approximately 30 minutes, Tip60 is very unstable. In normal conditions, the proteasome pathway permits to maintain low protein levels. Tip60 is ubiquitinated and targeted to proteasome-mediated degradation by Mdm2 but also by p300-associated E4 ubiquitin ligase. Tip60 is stabilized after DNA damage, and accumulates in cells. Moreover, Tip60 is the target of several post-translational modifications such as phosphorylation on serine 86 and 90 by cdc2 but also acetylation by p300/CBP acetyltransferases.

HTATIP (HIV-1 Tat interacting protein, 60kDa) Mattera L


Acetylation by p300/CBP occurs in the zinc finger of Tip60 but consequences of this modification are currently not known. Finally, a recent report shows that Tip60 is sumoylated at lysines 430 and 451 via Ubc9. No data are available about regulation of the Tip60 promoter.

Localisation Tip60 (Isoform 2) is nuclear. PLIP (Isoform 3) is nuclear but also cytoplasmic.

Function Tip60 is a Histone Acetyltransferase (HAT), which belongs to the MYST family. It participates in a multimolecular complex: The Tip60 complex, which contains proteins such as p400, Tip49a and Tip49b. Within this complex, Tip60 exerts its HAT activity on nucleosomal histone H4. Tip60 is involved in various cellular mechanisms: In transcription: Tip60 acts as a coactivator. Indeed, Tip60 is able to interact with transcription factors, such as E2F-1 or c-Myc. Tip60 can be recruited to Myc and E2F-1 target promoters and enhances Myc transactivation. It also acetylates histone H4 on several E2F responsive genes. Moreover Tip60 was found to be involved in nuclear receptor (NR) signaling and to be a NR-coregulator. In apoptosis and cell cycle arrest: Tip60 can interact with and acetylate the tumor suppressor p53. It enhances p53 binding to pro-apoptotic target genes like PUMA, Bax or Fas. Moreover, Tip60 is also required for cell growth arrest via the p21-dependent pathway. In DNA repair: Tip60 is involved in double strand breaks (DSB) repair. Interacting and acetylating ATM, Tip60 participates in DNA damage signaling. But, Tip60 is also involved directly in DSB repair since it is

recruited, with TRRAP, to the DSB site. Tip60 interacts with the chromatin surrounding sites of DSBs and this recruitment is responsible for hyperacetylation of histone H4.

Homology Tip60 CHROMO domain has been identified by sequence homology with the Heterochromatin-associated protein 1 (HP1) chromodomain, which recognizes methylated lysines. It also harbors the MYST domain, which is highly conserved from yeast to human. Homologs in other species: - S. Cerevisae: Esa1 - D. Melanogaster: DmeI/Tip60 - M. musculus: Htatip - R. norvegicus: Htatip Predicted: - P.troglodytes: HTATIP - M. mulatta: HTATIP

Mutations Note: No mutation in Tip60 protein has been currently described.

Implicated in Acquired Immunodeficiency Syndrome (AIDS) Disease Tip60 interacts with the HIV-1 transactivator Tat and this interaction inhibits Tip60 HAT activity. Moreover, in Jurkat cells, Tat enhances Tip60 turnover since it uses the p300/CBP-associated E4-type ubiquitin-ligase



activity to induce polyubiquitynation and degradation of Tip60. This targeting by Tat induces an impairment of Tip60-dependent apoptosis after DNA damage.

Neurodegenerative diseases: Alzheimer’s disease Disease In the nucleus of human H4 neuroglioma cells, TIP60 can interact with a free carboxyl-terminal intracellular fragment, APP-CT, which is generated by the cleavage of the Amyloid precursor protein APP by a gamma-secretase. This fragment induces apoptosis of neuroglioma and this cell death is enhanced when a wild type form of Tip60 is transfected. Thus Tip60 might play a role in Alzheimer’s disease neurodegeneration.

Spinocerebellar ataxia type-1 Disease TIP60 participates in a complex with ATXN1 and ROR-alpha in a conditional transgenic mouse model of Spinocerebellar ataxia type-1 (SCA1), one of the nine inherited polyglutamine neurodegenerative diseases.

Cancers: Prostate cancer Disease Immunohistochemistry experiments have shown that Tip60 accumulates in the nucleus of hormone-refractory prostate cancer compared to prostate hyperplasia and primary prostate cancer. Lung cancer and colon cancer Disease Real time RT-PCR experiments have shown that Tip60 mRNA is under expressed in colon and lung carcinomas.

Skin cancer Disease The expression levels of TIP60 protein, analyzed by western blot, were found to be greater in skin tumors as compared to adjacent non-tumor-bearing skin in a skin cancer mouse model (K6/ODC mouse). Additionally, the interaction between Tip60 and E2F1 is enhanced in these tumors.

HTLV-1 induced leukemogenesis Disease Enhancement of c-Myc transforming activity by HTLV-1 p30II oncoprotein in HeLa cells requires TIP60 HAT activity.

References Kamine J, Elangovan B, Subramanian T, Coleman D, Chinnadurai G. Identification of a cellular protein that

specifically interacts with the essential cysteine region of the HIV-1 Tat transactivator. Virology 1996;216(2):357-366.

Brady ME, Ozanne DM, Gaughan L, Waite I, Cook S, Neal DE, Robson CN. Tip60 is a nuclear hormone receptor coactivator. J Biol Chem 1999;274(25):17599-17604.

Creaven M, Hans F, Mutskov V, Col E, Caron C, Dimitrov S, Khochbin S. Control of the histone-acetyltransferase activity of Tip60 by the HIV-1 transactivator protein, Tat. Biochemistry 1999;38(27):8826-8830.

Ikura T, Ogryzko VV, Grigoriev M, Groisman R, Wang J, Horikoshi M, Scully R, Qin J, Nakatani Y. Involvement of the TIP60 histone acetylase complex in DNA repair and apoptosis. Cell 2000;102(4):463-473.

Ran Q, Pereira-Smith OM. Identification of an alternatively spliced form of the Tat interactive protein (Tip60), Tip60(beta). Gene 2000;258(1-2):141-146.

Gaughan L, Brady ME, Cook S, Neal DE, Robson CN. Tip60 is a co-activator specific for class I nuclear hormone receptors. J Biol Chem 2001;276(50):46841-46848.

Sheridan AM, Force T, Yoon HJ, O'Leary E, Choukroun G, Taheri MR, Bonventre JV. PLIP, a novel splice variant of Tip60, interacts with group IV cytosolic phospholipase A(2), induces apoptosis, and potentiates prostaglandin production. Mol Cell Biol 2001;21(14):4470-4481.

Gaughan L, Logan IR, Cook S, Neal DE, Robson CN. Tip60 and histone deacetylase 1 regulate androgen receptor activity through changes to the acetylation status of the receptor. J Biol Chem 2002;277(29):25904-25913.

Kinoshita A, Whelan CM, Berezovska O, Hyman BT. The gamma secretase-generated carboxyl-terminal domain of the amyloid precursor protein induces apoptosis via Tip60 in H4 cells. J Biol Chem 2002;277(32):28530-28536.

Legube G, Linares LK, Lemercier C, Scheffner M, Khochbin S, Trouche D. Tip60 is targeted to proteasome-mediated degradation by Mdm2 and accumulates after UV irradiation. EMBO J 2002;21(7):1704-1712.

McAllister D, Merlo X, Lough J. Characterization and expression of the mouse tat interactive protein 60 kD (TIP60) gene. Gene 2002;289(1-2):169-176.

Frank SR, Parisi T, Taubert S, Fernandez P, Fuchs M, Chan HM, Livingston DM, Amati B. MYC recruits the TIP60 histone acetyltransferase complex to chromatin. EMBO Rep 2003;4(6):575-580.

Halkidou K, Gnanapragasam VJ, Mehta PB, Logan IR, Brady ME, Cook S, Leung HY, Neal DE, Robson CN. Expression of Tip60, an androgen receptor coactivator, and its role in prostate cancer development. Oncogene 2003;22(16):2466-2477.

Legube G, Trouche D. Identification of a larger form of the histone acetyl transferase Tip60. Gene 2003;310:161-168.

Lemercier C, Legube G, Caron C, Louwagie M, Garin J, Trouche D, Khochbin S. Tip60 acetyltransferase activity is controlled by phosphorylation. J Biol Chem 2003;278(7):4713-4718.

Berns K, Hijmans EM, Mullenders J, Brummelkamp TR, Velds A, Heimerikx M, Kerkhoven RM, Madiredjo M, Nijkamp W, Weigelt B, Agami R, Ge W, Cavet G, Linsley PS, Beijersbergen RL, Bernards R. A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 2004;428(6981):431-437.

Legube G, Linares LK, Tyteca S, Caron C, Scheffner M, Chevillard-Briet M, Trouche D. Role of the histone acetyl transferase Tip60 in the p53 pathway. J Biol Chem 2004;279(43):44825-44833.

Taubert S, Gorrini C, Frank SR, Parisi T, Fuchs M, Chan HM, Livingston DM, Amati B. E2F-dependent histone acetylation



and recruitment of the Tip60 acetyltransferase complex to chromatin in late G1. Mol Cell Biol 2004;24(10):4546-4556.

Awasthi S, Sharma A, Wong K, Zhang J, Matlock EF, Rogers L, Motloch P, Takemoto S, Taguchi H, Cole MD, Lüscher B, Dittrich O, Tagami H, Nakatani Y, McGee M, Girard AM, Gaughan L, Robson CN, Monnat RJ Jr, Harrod R. A human T-cell lymphotropic virus type 1 enhancer of Myc transforming potential stabilizes Myc-TIP60 transcriptional interactions. Mol Cell Biol 2005;25(14):6178-6198.

Col E, Caron C, Chable-Bessia C, Legube G, Gazzeri S, Komatsu Y, Yoshida M, Benkirane M, Trouche D, Khochbin S. HIV-1 Tat targets Tip60 to impair the apoptotic cell response to genotoxic stresses. EMBO J 2005;24(14):2634-2645.

Sun Y, Jiang X, Chen S, Fernandes N, Price BD. A role for the Tip60 histone acetyltransferase in the acetylation and activation of ATM. Proc Natl Acad Sci USA 2005;102(37):13182-13187.

Hobbs CA, Wei G, Defeo K, Paul B, Hayes CS, Gilmour SK. Tip60 Protein Isoforms and Altered Function in Skin and Tumors that Overexpress Ornithine Decarboxylase. Cancer Res 2006;66(16):8116-8122.

Kim MS, Merlo X, Wilson C, Lough J. Co-activation of atrial natriuretic factor promoter by Tip60 and serum response factor. J Biol Chem 2006;281(22):15082-15089.

LLeonart ME, Vidal F, Gallardo D, Diaz-Fuertes M, Rojo F, Cuatrecasas M, López-Vicente L, Kondoh H, Blanco C, Carnero A, Ramón y Cajal S. New p53 related genes in human tumors: significant downregulation in colon and lung carcinomas. Oncol Rep 2006;16(3):603-608.

Murr R, Loizou JI, Yang YG, Cuenin C, Li H, Wang ZQ, Herceg Z. Histone acetylation by Trrap-Tip60 modulates loading of

repair proteins and repair of DNA double-strand breaks. Nat Cell Biol 2006;8(1):91-99.

Serra HG, Duvick L, Zu T, Carlson K, Stevens S, Jorgensen N, Lysholm A, Burright E, Zoghbi HY, Clark HB, Andresen JM, Orr HT. RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. Cell 2006;127(4):697-708.

Sykes SM, Mellert HS, Holbert MA, Li K, Marmorstein R, Lane WS, McMahon SB. Acetylation of the p53 DNA-binding domain regulates apoptosis induction. Mol Cell 2006;24(6):841-851.

Tang Y, Luo J, Zhang W, Gu W. Tip60-dependent acetylation of p53 modulates the decision between cell-cycle arrest and apoptosis. Mol Cell 2006;24(6):827-839.

Tyteca S, Vandromme M, Legube G, Chevillard-Briet M, Trouche D. Tip60 and p400 are both required for UV-induced apoptosis but play antagonistic roles in cell cycle progression. EMBO J 2006;25(8):1680-1689.

Cheng Z, Ke Y, Ding X, Wang F, Wang H, Ahmed K, Liu Z, Xu Y, Aikhionbare F, Yan H, Liu J, Xue Y, Powell M, Liang S, Reddy SE, Hu R, Huang H, Jin C, Yao X. Functional characterization of TIP60 sumoylation in UV-irradiated DNA damage response. Oncogene 2007 Aug 20;[Epub ahead of print].


Mattera L. HTATIP (HIV-1 Tat interacting protein, 60kDa). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):213-216.





HYAL1 (hyaluronoglucosaminidase 1) Demitrios H Vynios

Department of Chemistry, University of Patras, 26500 Patras, Greece


Online updated version: http://AtlasGeneticsOncology.org/Genes/HYAL1ID40903ch3p21.html DOI: 10.4267/2042/38523


Identity Hugo: HYAL1 Other names: EC 3.2.1.35; HYAL-1; NT6; LUCA1; LUCA-1; FUS2; Hyaluronidase-1 precursor; Hyaluronoglucosaminidase-1; Hs.75619; MGC45987 Location: 3p21.3 Local order: The gene of Hyal1 is tightly clustered with HYAL-2 and HYAL-3. The gene for Hyal-2, HYAL2, the earliest known lysosomal hyaluronidase, resides immediately centromeric to HYAL1. Note: The HYAL1 gene was identified as identical with LUCA-1, a candidate tumour suppressor gene, especially for tobacco-related cancers.

DNA/RNA Description The HYAL1 gene contains three exons and spans 12,492 bases (start: 50,312,324 bp to end 50,324,816 from 13pter) oriented at the minus strand.

Transcription Eight alternatively spliced transcript variants of this gene encoding six distinct isoforms have been described. The longest transcript has a length of 2,518 bps, however it is not translated to protein, since, by retaining intron 1 (occurring within exon 1), it has a number of stop codons. The longest transcript that produces active HYAL1 has a length of 2075 bps.

Pseudogene PHYAL1.

Protein Note: HYAL1 is a secreted somatic tissue hyaluronidase, and the predominant hyaluronidase in human plasma. Although HYAL1 is predominantly secreted, it has an acid pH optimum in vitro. HYAL1 can degrade high molecular weight hyaluronan to small oligomers, primarily to tetrasaccharides, whereas HYAL2 (the other major human hyaluronidase) high molecular mass hyaluronan to an approximately 20 kDa product (approximatively 50 saccharide units).

Description Size: 435 amino acids; Molecular mass: 48368 Da. The enzyme belongs to the group of carbohydrate-active enzymes (http://www.cazy.org/CASy), termed glycosyl hydrolase 56 family.

Expression HYAL1 is highly expressed in liver, kidney and heart and weakly expressed in lung, placenta and skeletal muscle. No expression is detected in adult brain. Isoform 1 is expressed only in bladder and prostate cancer cells, G2/G3 bladder tumor tissues and lymph node specimens showing tumor invasive tumors cells. Isoform 3, isoform 4, isoform 5 and isoform 6 are expressed in normal bladder and bladder tumor tissues. HYAL1 expression has been described in squamous cell carcinoma, in small cell lung cancer and glioma lines.

Localisation It is a secreted enzyme found in plasma and it is also present in lysosomes.

HYAL1 (hyaluronoglucosaminidase 1) Vynios DH


Function It is a hydrolytic enzyme (endo-beta-acetyl-D-hexosaminidase) with optimum pH about 3.7, acting on hyaluronan, chondroitin and chondroitin sulphate. It possesses also transglycosidase activity using hyaluronan and chondroitin sulphate or chondroitin as substrates. This reaction is not well understood, and the precise enzymatic mechanism is not known.

Homology The enzyme possesses 70-80% homology to orthologue hyaluronidases, 40% homology to paralogue hyaluronidases of the human and high homology with HYAL1 of other species.

Mutations Somatic There are not extended reports regarding mutations of HYAL1 gene. The patient with hyaluronidase deficiency was a compound heterozygote for two mutations in the HYAL1 gene: a 1412G-A mutation that introduced a nonconservative amino acid substitution (glu268 to lys) in a putative active site residue, and a complex intragenic rearrangement, 1361del37ins14, that resulted in a premature termination codon. In addition, the mutated HYAL1 gene has a markedly different expression pattern than the normal one.

Implicated in Mucopolysaccharidosis type IX (MPS9) Note: Defects in HYAL1 are the cause of mucopolysaccharidosis type IX, also called hyaluronidase deficiency.

Disease The clinical features are periarticular soft tissue masses, mild short stature and acetabular erosions, absence of neurological or visceral involvement and high hyaluronan concentration in the serum.

Cancer Note: HYAL1 is inactivated in most lung cancers in a conventional manner, by loss of heterozygosity or by homozygous deletion, at the DNA level. It is also inactivated in many head and neck carcinomas that are tobacco-related by aberrant splicing of the mRNA, so that only the nontranslatable form is transcribed. In addition, the expression of an alternative spliced isoform resulting in active enzyme may negatively regulate bladder tumor growth, infiltration, and angiogenesis. On the other hand, HYAL1 can function as oncogene in many cancers of the prostate and urinary tract and seems to play important role in squamous cell laryngeal carcinoma.

References Csóka AB, Frost GI, Wong T, Stern R. Purification and microsequencing of hyaluronidase isozymes from human urine. FEBS Lett 1997;417:307-310.

Frost GI, Csóka AB, Wong T, Stern R. Purification, cloning, and expression of human plasma hyaluronidase. Biochem Biophys Res Commun 1997;236:10-15.

Csóka AB, Frost GI, Heng HH, Scherer SW, Mohapatra G, Stern R. The hyaluronidase gene HYAL1 maps to chromosome 3p21.2-p21.3 in human and 9F1-F2 in mouse, a conserved candidate tumor suppressor locus. Genomics 1998;48:63-70.

Lepperdinger G, Strobl B, Kreil G. HYAL2, a human gene expressed in many cells, encodes a lysosomal hyaluronidase with a novel type of specificity. J Biol Chem 1998;273:22466-22470.

Csóka AB, Scherer SW, Stern R. Expression analysis of six paralogous human hyaluronidase genes clustered on chromosomes 3p21 and 7q31. Genomics 1999;60:356-361.

Triggs-Raine B, Salo TJ, Zhang H, Wicklow BA, Natowicz MR. Mutations in HYAL1, a member of a tandemly distributed multigene family encoding disparate hyaluronidase activities, cause a newly described lysosomal disorder, mucopolysaccharidosis IX. Proc Natl Acad Sci USA 1999;96:6296-6300.

Frost GI, Mohapatra G, Wong TM, Csóka AB, Gray JW, Stern R. HYAL1LUCA-1, a candidate tumor suppressor gene on chromosome 3p21.3, is inactivated in head and neck squamous cell carcinomas by aberrant splicing of pre-mRNA. Oncogene 2000;19:870-877.

Lerman MI, Minna JD. The 630-kb lung cancer homozygous deletion region on human chromosome 3p21.3: identification and evaluation of the resident candidate tumor suppressor genes. Cancer Res 2000;60:6116-6133.

Csóka AB, Frost GI, Stern R. The six hyaluronidase-like genes in the human and mouse genomes. Matrix Biol 2001;20:499-508. (Review).

Lokeshwar VB, Schroeder GL, Carey RI, Soloway MS, Iida N. Regulation of hyaluronidase activity by alternative mRNA splicing. J Biol Chem 2002;277:33654-33663.

Shuttleworth TL, Wilson MD, Wicklow BA, Wilkins JA, Triggs-Raine BL. Characterization of the murine hyaluronidase gene region reveals complex organization and cotranscription of Hyal1 with downstream genes, Fus2 and Hyal3. J Biol Chem 2002;277:23008-23018.

Junker N, Latini S, Petersen LN, Kristjansen PE. Expression and regulation patterns of hyaluronidases in small cell lung cancer and glioma lines. Oncol Rep 2003;10:609-616.

Jedrzejas MJ, Stern R. Structures of vertebrate hyaluronidases and their unique enzymatic mechanism of hydrolysis. Proteins 2005;61:227-238.

Lokeshwar VB, Cerwinka WH, Isoyama T, Lokeshwar BL. HYAL1 hyaluronidase in prostate cancer: A tumor promoter and suppressor. Cancer Res 2005;65:7782-7789.

Christopoulos TA, Papageorgakopoulou N, Theocharis DA, Mastronikolis NS, Papadas TA, Vynios DH. Hyaluronidase and CD44 hyaluronan receptor expression in squamous cell laryngeal carcinoma. Biochim Biophys Acta 2006;1760:1039-1045.

Lokeshwar VB, Estrella V, Lopez L, Kramer M, Gomez P, Soloway MS, Lokeshwar BL. HYAL1-v1, an Alternatively Spliced Variant of HYAL1 Hyaluronidase: A Negative Regulator of Bladder Cancer. Cancer Res 2006;66:11219-11227.

HYAL1 (hyaluronoglucosaminidase 1) Vynios DH


Stern R, Jedrzejas MJ. Hyaluronidases: their genomics, structures, and mechanisms of action. Chem Rev 2006;106:818-839. (Review).

Chao KL, Muthukumar L, Herzberg O. Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. Biochemistry 2007;46:6911-6920.


Vynios DH. HYAL1 (hyaluronoglucosaminidase 1). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):217-219.





MAML2 (mastermind-like 2) Kazumi Suzukawa, Jean-Loup Huret

Department of Hematology, Institute of Clinical Medicine, University of Tsukuba, Japan (KS); Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH)


Online updated version: http://AtlasGeneticsOncology.org/Genes/MAML2ID472.html DOI: 10.4267/2042/38524

This article is an update of: Stenman G. MAML2 (mastermind-like 2). Atlas Genet Cytogenet Oncol Haematol.2003;7(3):170-171. This work is licensed under a Creative Commons Attribution-Non-commercial-No Derivative Works 2.0 France Licence. © 2008 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Hugo: MAML2 Other names: hMam-3; KIAA1819 Location: 11q21

DNA/RNA Description Spans 365 kb; 5 exons.

Transcription A major transcript of 7.5 kb.

Protein Description 1153 aa, 125 kDa; conserved N-terminal basic domain (aa 29-92) which binds to the ankyrin repeat domain of Notch receptors; two acidic domains (aa 263-360 and 1124-1153) and a C-terminal transcriptional activation domain.

Expression Widely expressed.

Localisation Nuclear granules.

Function Mastermind-like coactivator for all four Notch receptors; forms a complex with the Notch intracellular domain (Notch ICD) and the CSL family of transcription factors (CSL: CBF1/RBP-jk, Suppressor of Hairless, LAG1), resulting in activation of the Notch

target genes HES1 and HES5; functions as a CSL-dependent transcriptional coactivator for ligand-stimulated Notch.

Homology MAML1 and MAML3.

Implicated in Mucoepidermoid carcinoma with t(11;19)(q21-22;p13) Disease - Most common type of malignant salivary gland tumor; - Second most frequent lung tumor of bronchial gland origin; - Rare tumour in the thyroid. The t(11;19) was found in samples from the three different sites.

Prognosis - Mucoepidermoid carcinomas have an unpredictable behaviour. - The CRTC1-MAML2 fusion transcript was found equally in low, intermediate and high grade tumours; however, tumours lacking the fusion transcript were significantly associated with metastases; they may represent a subset of aggressive tumours. - In another study, the median survival for fusion-positive patients was greater than 10 years compared to 1.6 years for fusion-negative patients.

Hybrid/Mutated Gene CRTC1-MAML2; exon 1 of CRTC1 fused to exons 2-5 of MAML2. Note: CRTC1 is also known as MECT1, or WAMTP1.

MAML2 (mastermind-like 2) Suzukawa K, Huret JL


Abnormal Protein CRTC1-MAML2. In the fusion protein, the first 171 aa including the basic domain of MAML2 are replaced by 42 aa of CRTC1; there are no sequence similarities in the N-terminal domains of MAML2 and CRTC1. The fusion protein activates transcription of the Notch target gene HES1 independently of both Notch ligand and CSL. Transforming activity of CRTC1-MAML2 fusion oncoprotein is mediated by mimicking constitutive activation of cAMP signaling, by activating CREB directly.

Warthin's tumor with t(11;19)(q21-22;p13) Note: In rare instances mucoepidermoid carcinoma may arise from or coexist with Warthin's tumors.

Disease Warthin's tumor is a salivary gland neoplasm consisting of benign epithelial and lymphoid components; malignant transformation is extremely rare.

Hybrid/Mutated Gene CRTC1-MAML2

Clear cell hidradenomas of the skin with t(11;19)(q21-22;p13) Disease Clear cell hidradenomas of the skin are benign sweat gland tumors of eccrine duct origin.

Hybrid/Mutated Gene CRTC1-MAML2; exon 1 of CRTC1 fused to exons 2 of MAML2.

inv(11)(q21q23) in therapy related leukemias Disease Therapy-related acute leukemia and MDS.

Hybrid/Mutated Gene MLL-MAML2; exon 1-7 of MLL fused to exons 2-5 of MAML2.

Abnormal Protein Hybrid transcript MLL/MAML2 contains the following domains: from MLL: AT-hook, DNA-Methyltransferase; from MAML2: Q rich domain, acidic domain.

To be noted Note: It is amazing that a similar fusion transcript (CRTC1-MAML2) can be seen both in a benign and in a malignant tumour of the same organ: Warthin's tumor, a benign salivary gland neoplasm, and mucoepidermoid carcinoma of the salivary gland: either another event differentiate the two, or the genetic event takes place in different cell types or in a given cell type at different states of differenciation. It has been hypothezised that CRTC1-MAML2 fusion is etiologically linked to benign and low-grade malignant tumors originating from diverse exocrine glands rather than being linked to a separate tumor entity.

References Bullerdiek J, Haubrich J, Meyer K, Bartnitzke S. Translocation t(11;19)(q21;p13.1) as the sole chromosome abnormality in a cystadenolymphoma (Warthin's tumor) of the parotid gland. Cancer Genet Cytogenet 1988;35(1):129-132.

Mark J, Dahlenfors R, Stenman G, Nordquist A. Chromosomal patterns in Warthin's tumor. A second type of human benign salivary gland neoplasm. Cancer Genet Cytogenet 1990;46(1):35-39.

Jhappan C, Gallahan D, Stahle C, Chu E, Smith GH, Merlino G, Callahan R. Expression of an activated Notch-related int-3 transgene interferes with cell differentiation and induces neoplastic transformation in mammary and salivary glands. Genes Dev 1992;6:345-355.

Nordkvist A, Gustafsson H, Juberg-Ode M, Stenman G. Recurrent rearrangements of 11q14-22 in mucoepidermoid carcinoma. Cancer Genet Cytogenet 1994;74:77-83.

MAML2 (mastermind-like 2) Suzukawa K, Huret JL


de la Pompa JL, Wakeham A, Correia KM, Samper E, Brown S, Aguilera RJ, Nakano T, Honjo T, Mak TW, Rossant J, Conlon RA. Conservation of the Notch signalling pathway in mammalian neurogenesis. Development 1997;124:1139-1148.

Stenman G, Petursdottir V, Mellgren G, Mark J. A child with a t(11;19)(q14-21;p12) in a pulmonary mucoepidermoid carcinoma. Virchows Archiv 1998;433; 579-581.

Wu L, Aster JC, Blacklow SC, Lake R, Artavanis-Tsakonas S, Griffin JD. MAML1, a human homologue of Drosophila Mastermind, is a transcriptional co-activator for NOTCH receptors. Nat Genet 2000;26:484-489.

Lin SE, Oyama T, Nagase T, Harigaya K, Kitagawa M. Identification of new human mastermind proteins defines a family that consists of positive regulators for notch signaling. J Biol Chem 2002;277:50612-50620.

Wu L, Sun T, Kobayashi K, Gao P, Griffin JD. Identification of a family of mastermind-like transcriptional coactivators for mammalian notch receptors. Mol Cell Biol 2002;22:7688-7700.

Tonon G, Modi S, Wu L, Kubo A, Coxon AB, Komiya T, O'Neil K, Stover K, El-Naggar A, Griffin JD, Kirsch IR, Kaye FJ. t(11;19)(q21;p13) translocation in mucoepidermoid carcinoma creates a novel fusion product that disrupts a Notch signaling pathway. Nat Genet 2003;33:208-213.

Enlund F, Behboudi A, Andrén Y, Oberg C, Lendahl U, Mark J, Stenman G. Altered Notch signaling resulting from expression of a WAMTP1-MAML2 gene fusion in mucoepidermoid carcinomas and benign Warthin's tumors. Exp Cell Res 2004;292(1):21-28.

Martins C, Cavaco B, Tonon G, Kaye FJ, Soares J, Fonseca I. A study of MECT1-MAML2 in mucoepidermoid carcinoma and Warthin's tumor of salivary glands. J Mol Diagn 2004;6(3):205-210.

Behboudi A, Winnes M, Gorunova L, van den Oord JJ, Mertens F, Enlund F, Stenman G. Clear cell hidradenoma of

the skin-a third tumor type with a t(11;19)--associated TORC1-MAML2 gene fusion. Genes Chromosomes Cancer 2005;43(2):202-205.

Wu L, Liu J, Gao P, Nakamura M, Cao Y, Shen H, Griffin JD. Transforming activity of MECT1-MAML2 fusion oncoprotein is mediated by constitutive CREB activation. EMBO J 2005;24(13):2391-2402.

Behboudi A, Enlund F, Winnes M, Andrén Y, Nordkvist A, Leivo I, Flaberg E, Szekely L, Mäkitie A, Grenman R, Mark J, Stenman G. Molecular classification of mucoepidermoid carcinomas-prognostic significance of the MECT1-MAML2 fusion oncogene. Genes Chromosomes Cancer 2006;45(5):470-481.

Nemoto N, Suzukawa K, Shimizu S, Shinagawa A, Takei N, Taki T, Hayashi Y, Kojima H, Kawakami Y, Nagasawa T. Identification of a novel fusion gene MLL-MAML2 in secondary acute myelogenous leukemia and myelodysplastic syndrome with inv(11)(q21q23). Genes Chromosomes Cancer 2007;46(9):813-819.

Tirado Y, Williams MD, Hanna EY, Kaye FJ, Batsakis JG, El-Naggar AK. CRTC1/MAML2 fusion transcript in high grade mucoepidermoid carcinomas of salivary and thyroid glands and Warthin's tumors: implications for histogenesis and biologic behavior. Genes Chromosomes Cancer 2007;46(7):708-715.

Winnes M, Mölne L, Suurküla M, Andrén Y, Persson F, Enlund F, Stenman G. Frequent fusion of the CRTC1 and MAML2 genes in clear cell variants of cutaneous hidradenomas. Genes Chromosomes Cancer 2007;46(6):559-563.


Suzukawa K, Huret JL. MAML2 (mastermind-like 2). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):220-222.





MUC16 (mucin 16, cell surface associated) Shantibhusan Senapati, Moorthy P Ponnusamy, Ajay P Singh, Maneesh Jain, Surinder K Batra

Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, 985870 Nebraska Medical Center, Durham Research center 7005, Omaha, NE 68198-5870, USA


Online updated version: http://AtlasGeneticsOncology.org/Genes/MUC16ID41455ch19q13.html DOI: 10.4267/2042/38525


Identity Hugo: MUC16 Other names: CA125; FLJ14303; Mucin-16 Location: 19p13.2 Note: MUC16 belongs to the subgroup of the membrane-anchored mucin. It is a type-1 glycopotein with heavy O- and N-type glycosylation.

DNA/RNA Description In the genome, MUC16 is localized in 19p13.2 chromosome and is coded by sequences present within

approximatively 179 kb of genomic DNA.

Transcription As per the present available information, there is a discrepancy regarding the total number of exons present in MUC16 genomic DNA. This discrepancy is due to the absence/presence of some of the genomic sequences (particularly for the repeat regions) in the available genomic databases. The terminal nine exons on both 5' and 3' ends code for the amino- and carboxy-terminal domains of MUC16, respectively. At the same time, it has been proposed that five consecutive exons code for a single repeat unit (SRU) of the central tandem repeat domain.

Shows the genomic organization of MUC16 gene.

Protein

Shows the structural organization of CA125/MUC16 protein.

MUC16 (mucin 16, cell surface associated) Senapati S, et al.


Description MUC16 protein harbors a central tandem repeat region, N-terminal domain and carboxy terminal domain. The N-terminal domain has 12070 numbers of amino acids rich in serine/threonine residues and accounts for the major O-glycosylation known to be present in CA125. The MUC16 protein back bone is dominated by tandem repeat region, which has more than 60 repeat domains, each composed of 156 amino acids. Though all the individual repeat units are not similar, most of them occur more than once in the sequence. The repeat units are rich in serine, threonine and proline residues, which are typical for any mucins. Each repeat unit has some homology to the SEA (Sea-urchin sperm protein, Enterokinase and Agrin) module, whose exact biological function is not known. The epitopes for known anti-CA125 antibodies (OC125 and M11) are thought to be present on a small cysteine ring region present in the tandem-repeat region of MUC16. The carboxy-terminal domain has 284 aminoacids and can be divided into three different regions: extra cellular, transmembrane and cytoplasmic tail. The extracellular part of the carboxy-terminal domain has many N-glycosylation sites and some O-glycosyaltion sites. Several in silico analyses suggest a putative cleavage site in the extracellular part of carboxy-terminal domain. The MUC16 cytoplasmic tail is 31 amino acids long and has many possible phosphorylation sites. The phosphorylation of CA125 in WISH cells has been reported by labeling with 32PO43- and immunoprecipitaion analysis but the exact site of phosphorylation is yet to be mapped. Interestingly, CA125 contains a putative tyrosine phosphorylation site (RRKKEGY), which was first recognized in Src family protein. This sequence is conserved in the translated mouse EST (AK003577) that has homology with CA125/MUC16 at the C-terminal end. Recently, it has been shown that MUC16 cytoplasmic tail, which contains a polybasic aminoacid sequence, interacts with cytoskeleton through ERM (ezrin/radixin/moesin) actin-binding proteins.

Expression The expression of MUC16 has been reported in human epithelia of conjunctiva, cornea, middle ear and trachea under normal physiological conditions. MUC16 is also expressed in ovarian carcinoma.

Localisation It is a type I membrane-bound protein and due to cleavage gets secreted into the extracellular space. On the ocular surface, MUC16 is expressed on the tips of the microplicae of the ocular surface.

Function MUC16 provides a disadhesive protective barrier to the ocular epithelial surface. Overexpression of CA125/MUC16 in ovarian cancer indicates its possible role in cancer pathogenesis. Studies have shown that CA125/MUC16 binds to mesothelin and galectin-1, which are overexpressed in ovarian cancer. It has also been shown that mesothelin-MUC16 interaction has significance in adhesion of ovarian cancer cells to mesothelial cells present on the inner wall of the peritoneum and on the surface of other abdominal organs. This cell to cell adhesion may help in ovarian cancer metastasis. It has been proposed that galectin-1 bound to MUC16 may cause apoptosis of T cells, and thus help in the suppression of the host immunity.

Homology Similar to mucin 16 of Pan troglodytes, Canis lupus familiaris, Mus musculus, Rattus norvegicus and Gallus gallus.

Implicated in Ovarian cancer Disease Epithelial ovarian cancer is the most lethal gynaecologic malignancy in the United States and other parts of the world. In the United States, ovarian cancer accounts for approximately 22,000 new cases and 16,000 deaths occurring every year. The epithelial ovarian carcinomas represent approximately 90% of all types of ovarian malignant neoplasms. Due to lack of specific signs and symptoms of this disease, coupled with lack of reliable screening strategies most patients are diagnosed in the advanced stage of the disease, resulting in low overall cure rates. Ovarian cancer patients are generally treated with surgical resection and subsequent platinum-based chemotherapy. Although, many patients initially respond well to chemotherapy, long term survival remains poor due to eventual tumor recurrence and emergence of drug-resistant disease. Overall, the five year survival rate is 45%.

Prognosis Since the last 20 years, CA125/MUC16 has been used as a well-established marker for diagnosis of ovarian cancer. It is mostly overexpressed in serous type of ovarian cancers and less likely to be expressed in mucinous tumors. More than 80% of ovarian cancer patients have elevated CA125 level during their treatment period. It has been shown that the disease progression is associated with an increase in serum CA125 level, while a decline in serum CA125 level is associated with response to therapy. In another finding,

MUC16 (mucin 16, cell surface associated) Senapati S, et al.


it has been shown that the trend of serum CA125 level during the first three courses of chemotherapy is a strong forecaster of re-examination findings in patients with ovarian carcinoma at the end of treatment. Interestingly, it has been shown that a normal CA125 level by the end of second or third chemotherapy is strongly linked to the survival of patients in stage 3 or stage 4 conditions. Also, variations in the CA125 value even within the normal range carry useful information regarding prediction of time to treatment failure. Additionally, in patients in stage 1 cancers it has been suggested that CA125 elevations are not related to the tumor mass volume. Recently, the potential of CA125/MUC16 as a therapeutic target has been harnessed by using an armed human antibody (3A5) against MUC16 protein.

Oncogenesis There is no experimental evidence in the scientific literature for a role of MUC16 in oncogenesis. However, MUC16 possesses many structural similarities with other membrane bound mucins, like MUC1 and MUC4, which are already shown to be functionally involved in different cancers. Transmembrane mucins are hypothesized to serve as sensors of the external environment and can transduce signals via the post-translational modifications of their cytoplasmic tail. Phosphorylation of MUC16 protein has already been reported. Though the exact interacting partner and the site of phosphorylation are unknown, the presence of potential phosphorylation sites in MUC16 cytoplasmic tail indicates the possible role of MUC16 in downstream signal transduction. Further, it has been shown that MUC16 interacts with galectin-1 and mesothelin and these interactions may have a role in cancer progression.

References O'Brien TJ, Beard JB, Underwood LJ, Dennis RA, Santin AD, York L. The CA 125 gene: an extracellular superstructure dominated by repeat sequences. Tumour Biol 2001;22(6):348-366.

Yin BW, Lloyd KO. Molecular cloning of the CA125 ovarian cancer antigen: identification as a new mucin, MUC16. J Biol Chem 2001;276(29):27371-27375.

O'Brien TJ, Beard JB, Underwood LJ, Shigemasa K. The CA 125 gene: a newly discovered extension of the glycosylated N-terminal domain doubles the size of this extracellular superstructure. Tumour Biol 2002;23(3):154-169.

Yin BW, Dnistrian A, Lloyd KO. Ovarian cancer antigen CA125 is encoded by the MUC16 mucin gene. Int J Cancer 2002;98(5):737-740.

Seelenmeyer C, Wegehingel S, Lechner J, Nickel W. The cancer antigen CA125 represents a novel counter receptor for galectin-1. J Cell Sci 2003;116(Pt 7):1305-1318.

Maeda T, Inoue M, Koshiba S, Yabuki T, Aoki M, Nunokawa E, Seki E, Matsuda T, Motoda Y, Kobayashi A, Hiroyasu F, Shirouzu M, Terada T, Hayami N, Ishizuka Y, Shinya N, Tatsuguchi A, Yoshida M, Hirota H, Matsuo Y, Tani K, Arakawa T, Carninci P, Kawai J, Hayashizaki Y, Kigawa T, Yokoyama S. Solution structure of the SEA domain from the murine homologue of ovarian cancer antigen CA125 (MUC16). J Biol Chem 2004;279(13):13174-13182.

Chauhan SC, Singh AP, Ruiz F, Johansson SL, Jain M, Smith LM, Moniaux N, Batra SK. Aberrant expression of MUC4 in ovarian carcinoma: diagnostic significance alone and in combination with MUC1 and MUC16 (CA125). Mod Pathol 2006;19(10):1386-1394.

Duraisamy S, Ramasamy S, Kharbanda S, Kufe D. Distinct evolution of the human carcinoma-associated transmembrane mucins, MUC1, MUC4 AND MUC16. Gene 2006;373:28-34.

Gubbels JA, Belisle J, Onda M, Rancourt C, Migneault M, Ho M, Bera TK, Connor J, Sathyanarayana BK, Lee B, Pastan I, Patankar MS. Mesothelin-MUC16 binding is a high affinity, N-glycan dependent interaction that facilitates peritoneal metastasis of ovarian tumors. Mol Cancer 2006;5(1):50.

Blalock TD, Spurr-Michaud SJ, Tisdale AS, Heimer SR, Gilmore MS, Ramesh V, Gipson IK. Functions of MUC16 in Corneal Epithelial Cells. Invest Ophthalmol Vis Sci 2007;48(10):4509-4518.

Chen Y, Clark S, Wong T, Chen Y, Chen Y, Dennis MS, Luis E, Zhong F, Bheddah S, Koeppen H, Gogineni A, Ross S, Polakis P, Mallet W. Armed antibodies targeting the mucin repeats of the ovarian cancer antigen, MUC16, are highly efficacious in animal tumor models. Cancer Res 2007;67(10):4924-4932. Erratum in Cancer Res 2007;67(12):5998.

Davies JR, Kirkham S, Svitacheva N, Thornton DJ, Carlstedt I. MUC16 is produced in tracheal surface epithelium and submucosal glands and is present in secretions from normal human airway and cultured bronchial epithelial cells. Int J Biochem Cell Biol 2007;39(10):1943-1954.


Senapati S, Ponnusamy MP, Singh AP, Jain M, Batra SK. MUC16 (mucin 16, cell surface associated). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):223-225.

Gene Section Review




MUC17 (mucin 17, cell surface associated) Wade M Junker, Nicolas Moniaux, Surinder K Batra

Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, 985870 Nebraska Medical Center, Durham Research Center 7005, Omaha, NE 68198-5870, USA


Online updated version: http://AtlasGeneticsOncology.org/Genes/MUC17ID41456ch7q22.html DOI: 10.4267/2042/38526


Identity Hugo: MUC17 Other names: MUC3; mucin-17; mouse Muc3; small intestinal mucin MUC3; membrane mucin 17; secreted mucin 17; intestinal membrane mucin MUC17 Location: 7q22.1 Note: Mucin glycoproteins are a diverse family of high molecular mass, heavily glycosylated proteins differentially expressed in epithelial tissues of the gastrointestinal, reproductive and respiratory tracts. Membrane mucins such as MUC17 are expressed by epithelial cells to provide protection, maintain luminal structure, and provide signal transduction. Often these molecules are over- or aberrantly expressed in cancers of epithelial origin. Mucins confer anti-adhesive properties in cancer cells that lose their apical/basal polarization. They also provide adhesive properties towards endothelial cells favoring dissemination of mucin expressing cancer cells. MUC17 is a recently fully characterized mucin that belongs to the membrane-bound subfamily of mucin (Moniaux et al., 2006). It is a cell surface glycoprotein that is found on epithelial cells in select tissues of the body (wade need to precise the know profile). Its structure consists of an extracellular domain that extends above the cell surface and an intracellular domain of 80 amino acid residues. New evidence suggests its de-regulation in pancreatic cancer (Moniaux et al., 2006; Moehle et al., 2006). The first partial length cDNA sequence, now known to correspond to MUC17, was identified by Van Klinken et al. (1997). At that time, van klinken isolated a chimeric cDNA clone overlapping MUC3 specific tandem repeat sequence and a new 59 aa sequence

repeated in tandem. This sequence was therefore believed to be part of MUC3 until Gum and collaborators identified the carboxy terminal sequence of the 59 aa tandem repeat and identified this sequence has part of a new mucin called MUC17. Indeed, in 2002, driven with the hypothesis that the 177 bp tandem repeated sequences were part of a new unidentified mucin, Gum et al. screened the public GenBankTM database and the proprietary Lifeseq Gold database (Incyte Genomics Inc., CA). By database searching and RT-PCR they extended the partial mucin sequence found during analysis of MUC3 and cloned a MUC17 fragment of 3,807 bp (accession number AF430017) (Gum et al., 2002). In 2006, Moniaux and Junker reported the complete coding sequence and organization of the MUC17 gene (Moniaux et al., 2006).

DNA/RNA Note: MUC17 was identified and localized to chromosome 7q22 by radiation hybridization mapping (Gum et al., 2002) where it resided in a gene cluster with mucins MUC3A, MUC3B, and MUC11/12 (Figure 1). These mucins reside next to each other and have function in intestinal epithelium integrity, but are expressed in other different tissues and likely have different functions as well. The mucin genes within the cluster are transcribed independent of one another. MUC17 shows a low degree of VNTR (variable number of tandem repeats) polymorphism with only three different genomic DNA sizes detected for the large tandemly repeated extracellular domain in 24 cancer lines (pancreas, colon, and breast) and in four healthy individuals control samples (Moniaux et al., 2006).

MUC17 (mucin 17, cell surface associated) Junker WM, et al.


Figure 1 - Chromosome location within the 7q22 mucin gene cluster. MUC17 resides in a gene cluster with mucins MUC3A, MUC3B, and MUC11/12 on chromosome 7 in the region q22.1. Upstream of the gene cluster 50 Kb is a potential open reading frame that shares similarity to MUC3. Approximately 26.7 Kb downstream of MUC17 reside TRIM56 and SERPINE1. Less that 1.2 Kb of genomic distance separates the 3' end of MUC12 from the beginning of MUC17. Figure 2 - MUC17 genomic and transcript bp size. The MUC17 gene encompasses (38,587 bp) 38.6 Kb of genomic sequence. The coding sequence contains 13 exons and is transcribed as a (14,360 bp) 14.4 Kb RNA that gives rise to full-length MUC17. The presence of an alternative splice site results in exclusion of exon 7, and produces a processed RNA that gives rise to a shorter MUC17/SEC.

Description Radiation hybridization mapping was performed using the GeneBridge4 radiation hybrid panel (Research Genetics, Huntsville, AL). Following hybridization, the MUC17 probe was detected in the panel of 93 independent human/hamster fusion clones with MUC17 specific primers, and data were analyzed using the GeneBridge4 server. This analysis indicated linkage to markers on chromosome 7q22 with an LOD score of 15. The presence or absence of MUC17 was concordant with STS (Sequence Tagged Site) marker D7S666 in 83 of 84 panel DNA samples that were unambiguous for both markers, thus positioning MUC17 near base 101,250,000 of chromosome 7 using the National Center for Biotechnology Information (NCBI) STS map. The NCBI electronic PCR server refined the interval to bases 98,871,000 and 99,054,000 of the STS map and positioned MUC17 approximately 2,000,000 bases telomeric of MUC3A on 7q22 (Gum et al., 2002). The MUC17 sequence has now been extended toward its 5'-extremity to complete the sequence and localize the promoter and regulatory elements. Rapid amplification of cDNA ends (RACE) and sequences from the Human Genome databases were used. The MUC17 gene is located within a 39-kb DNA fragment between MUC12 and SERPINE1 on chromosome 7 in the region q22.1 (Moniaux et al., 2006).

Transcription The MUC17 full-length coding sequence is transcribed as a (14,360 bp) 14.4 Kb mRNA encompassing 13 exons from a (38,587 bp) 38.6 kb genomic fragment (Figure 2). Alternate splicing generates two variants coding for a membrane-anchored and a secreted form of the protein (Moniaux et al., 2006) (Figure 3). The presence of an alternative splice event/site in the 3'-extremity of MUC17 was investigated by RT-PCR. RT-PCR was carried out on AsPC-1 cDNA. The generated amplification products were cloned into pCR® 2.1 and screened. Two distinct fragments were identified through sequencing. One of the fragments was 100% identical to the previous referenced sequence of MUC17 (accession number AJ606307). The second product revealed the occurrence of an alternative splice event that resulted in the skipping of exon 7. This alternative splice event generated a frame-shift which coded for the 21 (MUC17/SEC) specific amino acid residues and introduced a stop codon positioned 66 nucleotides after the junction. The resulting protein encodes a secreted form of MUC17 (accession number AJ606308), lacking the second EGF domain, the transmembrane domain and cytoplasmic tail (Figure 3). RT-PCR was carried out in four distinct cell lines, (pancreatic AsPC-1, and colonic LS174T, CaCo-2, and Ls180), representing different tissues, i.e. pancreas and colon (Moniaux et al., 2006). Two



amplification products were detected. Sequencing of the major amplification product identified it as the MUC17 sequence described by Gum et al. (accession number AF430017), while the other amplicon (minor) corresponded to an alternatively spliced variant (skipping of exon 7), the secreted form of MUC17, referred to as MUC17/SEC. The level of expression of MUC17/SEC seemed very low in the cell lines investigated; the intensity of the corresponding band was very faint as compared to the MUC17 fragment (Moniaux et al., 2006). MUC17 is expressed in select cell lines including pancreatic AsPC-1 and HPAF-II; and colon cancer cell lines LS174T, Caco-2, NCI-H498, and HM3 (Gum et al., 2002; Moniaux et al., 2006). Tissue expression was first shown by RNA dot blot analysis (Clontech multiple tissue expression blot). MUC17 is expressed in intestinal tissues, with the highest levels found in the duodenum (highest level) and in the transverse colon (85% of the level detected in duodenum). The only non-intestinal tissues found to express MUC17 in this analysis were stomach and fetal kidney. Both tissues showed approximately 7% of the expression level detected in the duodenum (Gum et al., 2002). In-situ hybridization was conducted to determine the cell specificity of MUC17 expression in the small intestine. In-situ hybridization showed MUC17 expression predominantly in the apical region of villi absorptive cells. Barely detectable expression was found in immature cells of the crypts. No expression was detected in goblet cells (Gum et al., 2002). Therefore, MUC17 expression is localized to the mature, absorptive cells of intestinal villi epithelium and is expressed in pancreatic cancer tissue (Moniaux et al., 2006). The MUC17 gene is located within a 39 kb DNA fragment between MUC12 and SERPINE1 on chromosome 7. Approximately 1.2 Kb of sequence lies between the 3' end of MUC12 and the 5' UTR of the MUC17 gene. Expression of MUC17 is regulated by this 1,146-bp fragment upstream of MUC17 which contains various VDR/RXR, GATA, NFkB, and Cdx-2 response elements. Like mouse Muc3 (mMUC3), regulation of expression is controlled by both growth factors and cytokines (unpublished data, Junker and Batra, 2007). The mMuc3 promoter has consensus binding sites for AP1, CREB, SP1, NF kappa B, GATA binding protein, and Cdx. Reporter constructs demonstrate that IL-4, IL-6, EGF, and PMA increase mMuc3 promoter activity 35-58% of control levels. TNF-a and IFN-g showed a lesser degree of stimulation (Shekels et al., 2003). Regulation of mMuc3 by cytokines and growth factors suggests an active role of mouse Muc3 and its human homologue, MUC17 in intestinal mucosal defense. A recent study conducted by Moehle et al. (2006) concerned the aberrant expression and allelic variants of mucin genes associated with inflammatory bowel

disease (IBD). The aim of the study was to characterize changes in the expression profiles of genes related to intestinal epithelial function by comparing biopsy samples from patients with Ulcerative Colitis (UC) and Crohn's disease (CD), to controls; as the loss of intestinal mucosal integrity is an important factor in IBD. DNA-microarray analysis was applied and showed that mucin genes are differentially regulated in CD and UC. The loss of intestinal integrity is an important factor in the pathogenesis of inflammatory bowel disease. A coordinate down regulation of mucins was observed in a pool of biopsy RNAs (n=4) taken from affected and unaffected (control) regions of the terminal ileum and colon of CD and UC patients. No expression in the biopsy samples was detected for MUC6, MUC7, MUC8, MUC9, MUC11, MUC15, MUC16, and MUC18. Highest mucin expression values were displayed my MUC2, MUC13, and MUC17 in the ileum and the colon, while MUC12 was expressed in the colon. The relative expression levels for MUC1, MUC2, MUC4, MUC5B, MUC12, MUC13, MUC17, and MUC20 showed strong down regulation with decrease factors ranging from -1.3 to -48.5 fold. MUC17 showed a -4.3 and -2.6 fold decrease in Crohn’s disease and Ulcerative Colitis respectively in the colon, but showed an apparent increase of 1.2 and 1.1 fold respectively in the same diseases in the ileum biopsy pooled RNAs. Real-time RT-PCR TaqMan assays were conducted to obtain a specific overview of mucin expression in human tissues. An initial analysis of nine mucin genes in a panel of 26 different tissues was completed. The authors confirmed the relative expression values (average intensity) of DNA-microarray data (i.e., higher expression levels of MUC4, MUC5B, MUC12, and MUC20 were obtained in the colon compared to the ileum RNA; MUC17 was expressed higher in the ileum than in the colon). A meta-analysis of 11 genome-wide linkage studies for IBD revealed 38 significant IBD loci (Brant et al., 2004). Interestingly, all mucin family member loci reside within or directly beside these IBD candidate loci. Therefore, Moehle et al. performed allelic discrimination of one candidate exonic SNP within each mucin gene in UC (n=220), CD (n=181), and control (n=250) patient samples. Significant associations were detected for the MUC2, MUC4, and MUC13 mucin SNPs. The von Wildebrand Factor (vWF) domain of MUC2 (11p15, A/G, V116M) is associated with CD, the vWF domain of MUC4 (3q29, G/T, A585S) with UC, and the cytoplasmic tail domain of MUC13 (3q13.1, A/C, R502S) with UC. Unassessed SNPs in the mucin genes may still be associated and remain unchecked. The abundance of NFkB sites present in mucin promoters prompted the authors (Moehle et al., 2006) to explore regulation of mucin expression in the Ls174T colon cell line model. The ligands TNF-a,



TGFb, and LPS induced expression of MUC17 as assayed by quantitative real-time PCR. TNF-a was able to induce a time-dependent up-regulation of all the monitored mucin genes (MUC-1, -2, MUC-5AC, -5B, -12, -13, -17, -20). All monitored mucin genes showed increased expression with TGFb treatment. Cooperation of NFkB with TGFb signaling has been reported (Jono et al., 2002). Co-incubations of TGFb with NFkB pathway inhibitors (CAPE and MG132) resulted in a decrease of mucin expression, thus demonstrating a link between these two pathways with regards to mucin gene regulation. Treatment of the Ls174T colon cancer cell line with sodium butyrate had little or no effect on induction of MUC17 expression level but did induce expression of MUC3 which is not highly expressed in the cell line (Gum et al., 2002). Similarly, incubation of MUC17 non-expressing cell line, MiaPaCa, with HDAC inhibitor 5-aza-cytosine had little effect if any on MUC17 expression.

Protein Note: 4493 aa; 425.5 kDa (note: without modification such as glycosylation)

Description MUC17 is classified as a membrane-bound mucin glycoprotein. The protein may serve as a cellular receptor. The deduced full-length membrane-bound amino acid sequence (4493 aa) shows the presence of various mucin domains (Figure 3). A signal sequence

targets the protein to the plasma membrane. The majority of the molecule encodes the mucin central domain that is modified extensively by glycosylation and is displayed on the extracellular face of the cell. The central domain of MUC17 contains 63 repeats of 59 amino acid sequence (177 bp) that are repeated in tandem. This tandem repeat 'central domain' is followed by a region of unique degenerate tandem repeats and mucin-like sequences (i.e. that are repetitive, G/C rich, and contain a high content of threonine, serine, and proline amino acids). Two EGF-like domains flank both sides of a SEA module and precede the transmembrane domain. Putative N-glycosylation sites occur near the carboxyl terminus. The 80 amino acid C-terminal cytoplasmic domain has potential serine and tyrosine phosphorylation sites.

Expression RNA blot analysis and RT-PCR suggests MUC17 is expressed in the digestive tract, primarily in the duodenum (highest level) and the transverse colon (85% of the level detected in duodenum) (Gum et al., 2002). Expression is also reported in the terminal ileum (Moehle et al., 2006) with MUC17 expressed higher in the ileum than in the colon (quantitative RT-PCR, microarray analysis). Many colon and pancreatic cancer cell lines do express varied levels of MUC17 (Gum et al., 2002; Moniaux et al., 2006). An over-expression of MUC17 by Western blot and immunohistochemical analyses in pancreatic tumor cell lines and tumor tissues compared to normal pancreas samples is seen (Moniaux et al., 2006).

Figure 3 - full length MUC17 and secreted MUC17/SEC. Full-length MUC17 contains a 25 amino acid leader peptide (secretion, membrane targeting signal), a central domain with tandemly repeated sequence, two EGF-like domains, a SEA domain, transmembrane domain, and an 80 amino acid cytoplasmic tail domain. Usage of an alternative splice site excludes exon 7 and introduces a frame shift that creates a premature stop codon 66 nucleotides after the splice junction, within the SEA domain coding sequence. This shorter transcript results in a secreted form of the protein, MUC17/SEC, which has 21 unique C-terminal amino acids, and lacks the second EGF domain, transmembrane domain, and C-terminal cytoplasmic tail.



Localisation Surface localization of the smaller subunit of MUC17 is reported to be dependent on its N-glycosylation status (Ho et al., 2003). MUC17 contains a SEA domain, a transmembrane domain, and putative N-glycosylation sites in the carboxyl terminus. Mucins that possess a SEA domain usually undergo an auto-proteolytic cleavage event within the domain (Macao et al., 2006) to yield two subunits, the smaller of which is associated with the surface membrane. Ho and colleagues reported that the ASPC-1 pancreatic cancer cell line shows three main bands (38, 45, and 49 kDa) of immunoreactivity with an antibody directed against a site downstream of the postulated SEA cleavage site (Ho et al., 2003). Treatment with N-glycan specific hydrolases showed the 38 kDa band contained high mannose glycans, whereas the 45 and 49 kDa bands contained complex-type glycans. Surface biotinylation studies revealed that only forms possessing complex-type N-glycans were localized to the cell surface. Both tunicamycin (N-glycosylation inhibitor) and brefeldin A (an inhibitor of protein transport) reduced surface localization. Surface localization of the smaller subunit of MUC17 therefore appears to be dependent on its N-glycosylation status in AsPC-1 pancreatic cancer cells. Immunohistochemical analysis of mouse Muc3 (the homologue of MUC17) revealed strong staining in goblet cells and patchy staining of surface columnar cells in the duodenum, small intestine, caecum, colon and rectum (Shekels et al., 1998). Northern blot analysis indicates that the mRNA is approximately 13.5 kb. Highest expression was detected in the caecum with lesser amounts detected in the colon and small intestine. No message was found in mouse stomach, trachea, lung, kidney, esophagus or pancreas (Shekels et al., 1998). In-situ hybridization studies show expression at the tips of villi, in the upper crypts, and in surface cells of the caecum and colon (Shekels et al., 1998). Rodent (mouse) mMuc3 and (rat) rMuc3 are assumed to represent secretory mucins expressed in columnar and goblet cells of the intestine. In-situ hybridization with a 3'-probe localized (rat) rMuc3 expression generally to columnar cells. Two antibodies specific for the C-termini of rat rMuc3 localized the protein to apical membranes and cytoplasm of columnar cells. An antibody to the tandem repeat sequence, however, localized the protein to both columnar and goblet cells. Cesium chloride ultracentrifugation was used to isolate both light- and heavy-density fractions. A full-length membrane-associated form (light density) was found in columnar cells, whereas, the carboxyl-truncated soluble form of rat Muc3 (heavy density) was present in goblet cells (Wang et al., 2002). A similar localization of human MUC17 may exist as the splice variant, MUC17SEC, encodes a truncated protein that is missing the second EGF-like domain, the

transmembrane domain, and the cytoplasmic tail. The MUC17/SEC protein is believed to be a soluble protein as the absence of the transmembrane domain would result in its secretion from the cell. In a mouse model of human cystic fibrosis, both soluble Muc3 and goblet cell Muc2 are increased and hyper-secreted contributing to the excess intestinal mucus of cystic fibrosis mice (Khatri et al., 2001).

Function MUC17 is a membrane-bound glycoprotein that may serve as a cellular receptor through its extended, repetitive extracellular glycosylation domain. The extracellular domain may serve a lubricant functionality and provide a signal transduction capability. Membrane mucins, such as MUC17, function in epithelial cells to provide cytoprotection, maintain luminal structure, provide signal transduction, and confer anti-adhesive properties to cancer cells which lose their apical/basal polarization. Outside-in signaling may be transduced by the protein's interaction with extracellular matrix constituents including growth factors and cytokines and/or potentially binding of Ca2+ or EGF ligands to extracellular displayed EGF-like domains. The 80 amino acid cytoplasmic domain has potential serine and tyrosine phosphorylation sites to convey extracellular signals. Analysis of mouse Muc3 showed that a definitive proteolytic cleavage occurs during processing in the endoplasmic reticulum. Recombinant products consisted of a V5-tagged 30 kDa extracellular glycopeptide and a Myc-tagged 49 kDa membrane-associated glycopeptide. Throughout their cellular transport to the plasma membrane, the two fragments remained associated by non-covalent SDS-sensitive interactions. Site-specific mutagenesis showed requirement for glycine and serine residues in the cleavage sequence Leu-Ser-Lys-Gly-Ser-Ile-Val-Val, which is found in the SEA domain between the two EGF-like motifs of the mucin. A similar cleavage sequence has been reported in human MUC1 and analogous sites are present in human MUC3, MUC12, MUC16 and MUC17. Proteolytic cleavage may be a conserved characteristic of the membrane-bound mucins, and possibly precedes the release of their large extracellular domains at cell surfaces (Wang et al., 2002).

Homology The similarity of human MUC17 to rodent Muc3 (mouse and rat) was first reported by JR Gum Jr and colleagues. This similarity was represented by a cladogram calculated using sequences initiating at the start of the second EGF-like domain and continuing through to the carboxy-termini of the proteins. The analysis suggests that MUC17 diverged from human MUC3 earlier in evolution than the divergence of primates and rodents, and suggests that MUC17 is the



true structural homolog of rodent Muc3 (Gum et al., 2002). Whether MUC17 is the functional homolog of rodent Muc3 is still unclear, however, and needs to be experimentally proven. Chromosome computer analysis assigns mouse Muc3 to mouse chromosome 5, a region of synteny to human chromosome 7, the location of the human MUC3, MUC12, and MUC17 mucin genes. NCBI HomoloGene reports MUC17 is conserved in Coelomata or in organisms higher than coelenterates and certain primitive worms. Expression of the mouse Muc3 mucin has been characterized in terms of regulation of its promoter by cytokines and growth factors. Mouse Muc3 is now believed to be the true structural homologue of human MUC17 due to higher sequence similarity to MUC17

than human MUC3 (Moniaux et al., 2006). The N-terminal domain for MUC17 is coded by two exons, whereas for MUC3, it is coded by a single exon. Gum and colleagues (2002) showed that the degree of sequence homology between the carboxy-extremity of MUC17 and mMuc3 was higher than that between MUC3 and mMUC3. An alignment of the amino-extremities of MUC17, MUC3, and mMuc3 is shown. No similarity is shown by MUC3 and mMuc3, but a high degree of identity exists between MUC17 and mMUC3. Their similar structural organization and high degree of identity show that MUC17 is the human homologue of mMuc3. A search of the National Center for Bioinformatics HomoloGene and UniGene databases returned the following suggested sequences for comparison.

HomoloGene:88635. Gene conserved in Coelomata

Human H. sapiens MUC17 mucin 17, cell surface associated, chromosome 7q22.1, GeneID: 140453

Chimpanzee (West African) P. troglodytes MUC17 mucin 17, chromosome 7, GeneID: 740201

Mouse M. musculus Muc3 mucin 3, intestinal

Rhesus monkey Macaca mulatta chromosome 3

Human Homo sapiens CAE54435 membrane mucin MUC17 (Homo sapiens) gi/51869309/emb/CAE54435.1/(51869309)

Human Homo sapiens CAE54436 secreted mucin MUC17 (Homo sapiens) gi/51869311/emb/CAE54436.1/(51869311)

Human Homo sapiens AAL89737 intestinal membrane mucin MUC17 (Homo sapiens) gi/19526645/gb/AAL89737.1/AF430017_1(19526645)

Human Homo sapiens AAI26316 MUC17 protein (Homo sapiens) gi/118835615/gb/AAI26316.1/(118835615)

Human Homo sapiens NP_001035194 mucin 17 (Homo sapiens) gi/91982772/ref/NP_001035194.1/(91982772)

Chimpanzee (West African) Pan troglodytes XP_001142083 PREDICTED: mucin 17 (Pan troglodytes)

gi/114615083/ref/XP_001142083.1/(114615083)

Opossum (gray short-tailed)

Monodelphis domestica XP_001375221

PREDICTED: similar to membrane mucin MUC17 (Monodelphis domestica) gi/126323716/ref/XP_001375221.1/(126323716)


Monodelphis domestica

XP_001371346 PREDICTED: similar to MUC17 protein (Monodelphis domestica) gi/126309309/ref/XP_001371346.1/(126309309)

Chicken Gallus gallus XP_001233667 PREDICTED: similar to intestinal membrane mucin MUC17, partial (Gallus gallus) gi/118121828/ref/XP_001233667.1/(118121828)

Mosquito Aedes aegypti EAT36316 secreted mucin MUC17, putative (Aedes aegypti), gi/108872091/gb/EAT36316.1/(108872091)


Monodelphis domestica LOC100017948

similar to MUC17 protein (Monodelphis domestica) , Chromosome: 2, GeneID: 100017948

Chicken Gallus gallus LOC770330 similar to intestinal membrane mucin MUC17 (Gallus gallus), Chromosome: Un, GeneID: 770330


Monodelphis domestica

LOC100023779 similar to membrane mucin MUC17 (Monodelphis domestica), Chromosome: 3, GeneID: 100023779

Chimpanzee (West African) Pan troglodytes LOC463614 similar to membrane mucin MUC17 (Pan troglodytes), Chromosome: 7,

GeneID: 463614

Rat (Brown Norway) Rattus norvegicus Muc17_predicted

Muc17_predicted and Name: mucin 17 (predicted) (Rattus norvegicus), Chromosome: 7, GeneID: 295035

Fruit Fly D. melanogaster Sgs1 Salivary gland secretion 1, 25B2-3 puff, salivary glands, third instar



Selected Protein Similarities Comparison of sequences in UniGene with proteins supported by a complete genome. The alignments can suggest function of a gene.

C. elegans 32.92 % / 636 aa ref:NP_505150.1 - reverse transcriptase (Caenorhabditis elegans)

S. cerevisiae 30.55 % / 644 aa pir:S48478 - S48478 glucan 1,4-alpha-glucosidase

R. norvegicus 27.24 % / 647 aa pir:A53577 - A53577 ascites sialoglycoprotein 1 - rat

M. musculus 24.77 % / 633 aa ref:NP_035871.1 - zonadhesin (Mus musculus)

A. thaliana 23.94 % / 531 aa ref:NP_564694.1 - expressed protein (Arabidopsis thaliana)

D. melanogaster 22.31 % / 637 aa sp:Q02910 - CPN_DROME CALPHOTIN

E. coli 18.05 % / 563 aa ref:NP_287395.1 - putative membrane protein of prophage CP-933X (Escherichia coli O157:H7 EDL933)

Implicated in Pancreatic adenocarcinoma Disease Worldwide, pancreatic cancer is the eleventh most common cancer. In the United States of America, pancreatic cancer is the fourth leading cause of cancer related death. Pancreatic cancer presents a 5-year survival rate of just 5%. The incidence and age-adjusted mortality rate (approximatively 95%) are almost equal, underscoring the aggressive nature of the disease.

Prognosis Currently, no approved diagnostic biomarker for pancreatic cancer is licensed in the United States. The CA19-9 mucin epitope is used for the diagnosis of ovarian cancer and other mucins are being developed for the detection of breast cancer (MUC1) and pancreatic cancer (MUC1, MUC4, MUC3, MUC17). The DUPAN-2 antibody recognizes a tumor-associated antigen carried by the MUC4 protein and is used as a clinical diagnostic for pancreatic adenocarcinoma in Japan. MUC4 is aberrantly expressed in 80% of pancreatic adenocarcinomas and is not expressed in the normal pancreas or benign pancreatitis. In addition MUC4 is expressed early in the onset of pancreatic cancer (detected in pancreatic intraepithelial neoplasia (PanIN) stage I disease). Similarly, MUC17 is aberrantly expressed in pancreatic adenocarcinomas as compared to no expression in the normal pancreas or benign pancreatitis. MUC17 is expressed early in the progression of pancreatic cancer with localization to well defined pancreatic ductal structures undergoing malignant transformation (Moniaux et al., 2006).

Oncogenesis In an in vivo model, subcutaneous injection of MUC17 AsPC-1 knock-down cells show slight increase in tumorigenicity in relation to parental control cells; although no significant difference was detected due to large variation in tumor weights. A xenograph model with knock-down cell lines injected orthotopically into the pancreas head showed an increased potential to metastasize to the peritoneal cavity and increased

tumor mass (weight) in relation to mice injected with the parental cell line (control group) transfected with a scrambled RNAi sequence (unpublished data, Junker and Batra, 2007).

To be noted Note: Conserved in Coelomata. From (HomoloGene: 88635. Gene conserved in Coelomata). Coelome: the cavity within the body of all animals higher than the coelenterates and certain primitive worms, formed by the splitting of the embryonic mesoderm into two layers. In mammals, the coelome forms the peritoneal, pleural, and pericardial cavities.

References Van Klinken BJ, Van Dijken TC, Oussoren E, Buller HA, Dekker J, Einerhand AW. Molecular cloning of human MUC3 cDNA reveals a novel 59 amino acid tandem repeat region. Biochem Biophys Res Commun 1997;238(1):143-148.

Shekels LL, Hunninghake DA, Tisdale AS, Gipson IK, Kieliszewski M, Kozak CA, Ho SB. Cloning and characterization of mouse intestinal MUC3 mucin: 3' sequence contains epidermal-growth-factor-like domains. Biochem J 1998;330:1301-1308.

Khatri IA, Ho C, Specian RD, Forstner JF. Characteristics of rodent intestinal mucin Muc3 and alterations in a mouse model of human cystic fibrosis. Am J Physiol Gastrointest Liver Physiol 2001;280:G1321-1330.

Gum JR Jr, Crawley SC, Hicks JW, Szymkowski DE, Kim YS. MUC17, a novel membrane-tethered mucin. Biochem Biophys Res Commun 2002;291(3):466-475.

Jono H, Shuto T, Xu H, Kai H, Lim DJ, Gum JR Jr, Kim YS, Yamaoka S, Feng XH, Li JD. Transforming growth factor-beta -Smad signaling pathway cooperates with NF-kappa B to mediate nontypeable Haemophilus influenzae-induced MUC2 mucin transcription. J Biol Chem 2002;277(47):45547-45557.

Wang R, Khatri IA, Forstner JF. C-terminal domain of rodent intestinal mucin Muc3 is proteolytically cleaved in the endoplasmic reticulum to generate extracellular and membrane components. Biochem J 2002;366:623-631.

Ho JJ, Jaituni RS, Crawley SC, Yang SC, Gum JR, Kim YS. N-glycosylation is required for the surface localization of MUC17 mucin. Int J Oncol 2003;23:585-592.

Shekels LL, Ho SB. Characterization of the mouse Muc3 membrane bound intestinal mucin 5' coding and promoter regions: regulation by inflammatory cytokines. Biochim Biophys Acta 2003;1627:90-100.



Brant SR, Shugart YY. Inflammatory bowel disease gene hunting by linkage analysis: rationale, methodology, and present status of the field. Inflamm Bowel Dis 2004;10(3):300-311. (Review).

Macao B, Johansson DG, Hansson GC, Hard T. Autoproteolysis coupled to protein folding in the SEA domain of the membrane-bound MUC1 mucin. Nat Struct Mol Biol 2006;13(1):71-76.

Moehle C, Ackermann N, Langmann T, Aslanidis C, Kel A, Kel-Margoulis O, Schmitz-Madry A, Zahn A, Stremmel W, Schmitz G. Aberrant intestinal expression and allelic variations of mucin

genes associated with inflammatory bowel disease. J Mol Med 2006;84(12):1055-1066.

Moniaux N, Junker WM, Singh AP, Jones AM, Batra SK. Characterization of human mucin MUC17. Complete coding sequence and organization. J Biol Chem 2006;281:23676-23685.


Junker WM, Moniaux N, Batra SK. MUC17 (mucin 17, cell surface associated). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):226-233.

Gene Section Review




PTHLH (parathyroid hormone-like hormone) Sai-Ching Jim Yeung

The University of Texas M. D. Anderson Cancer Center, Department of General Internal Medicine, Ambulatory Treatment and Emergency Care, Department of Endocrine Neoplasia and Hormonal Disorders, 1515 Holcombe Boulevard, Unit 437, Houston, Texas 77030, USA


Online updated version: http://AtlasGeneticsOncology.org/Genes/PTHLHID41897ch12p11.html DOI: 10.4267/2042/38527


Identity Hugo: PTHLH Other names: PTHLP (parathyroid hormone-like protein); PTHRP (parathyroid hormone-related protein); PTHrP; PTH-rP (PTH-related protein); PTHR; HHM (humoral hypercalcemia of malignancy); Osteostatin; PLP (parathyroid-like protein); MGC14611 Location: 12p11.22

DNA/RNA Description PTHLP is encoded by a single gene that is highly conserved among species. The gene is composed of 7 exons spanning a region of 13,899 bases (Start:

28,002,284 bp from pter; End: 28,016,183 bp from pter). Orientation: minus strand. The genomic DNA for the PTHLP gene was isolated from a human placental genomic library.

Transcription The sequence is supported by 3 sequences from 3 cDNA clones.

Pseudogene None.

Protein Description Size: 177 amino acids, 20194 Da.

This diagram represents schematically one possible proteolytic processing pattern of PTHLP into 3 bioactive peptides. The mid-region of PTHLP contains the nuclear localization signal (NLS).

PTHLH (parathyroid hormone-like hormone) Yeung SCJ


The PTHLP gene has seven exons, and its transcripts are processed by alternative splicing into three isoforms, encoding proteins with 139, 173 and 141 amino acids. The pattern of expression of PTHLP mRNA isoforms may be cell type-specific. Although different tumors may have different PTHLP splicing patterns, there are no tumor-specific transcripts. PTHLP is processed into a set of distinct peptide hormones by endoproteolytic cleavage of the initial translation products: mature N-terminal, mid-region and C-terminal secretory peptides, each having its own distinct function. The distribution of the endopeptidase processing enzymes (PTP (prohormone thiol protease), prohormone convertases 1 and 2 ( PC1 and PC2 )) may vary in different tissues. PTP cleaved the PTHLP precursor at the multibasic, dibasic, and monobasic residue cleavage sites to generate the NH2-terminal peptide (residues 1-37, having PTH-like and growth regulatory activities), the mid-region domain (residues 38-93, regulating calcium transport and cell proliferation), and the COOH-terminal domain (residues 102-141, modulating osteoclast activity).

Expression PTHLP is a protein polyhormone produced by most if not all tissues in the body. It is secreted during both fetal and postnatal life. Although PTHLP is found in the circulation, most of its activity appears to be paracrine. A complex of transcription factors and coactivators (CREB, Etsl and CBP) regulates PTHLP transcription and may contribute to the alterations associated with the promotion of carcinogenesis. Disruption of the normal regulation during cancer progression may in part be associated with TGF-beta1 -induced changes in PTHLP mRNA isoform expression and stability. TGF-beta activates PTHLP expression increasing transcription from the P3 promoter through a synergistic interaction of Smad3 and Ets1. ERK1/ERK2 -dependent Ets2/PKCepsilon synergism also appears to regulate PTHLP expression in breast cancer cells. The PTHLP gene is also under the transcriptional control of glucocorticoids and vitamin D. 1,25-dihydroxy vitamin D3 treatment increases PTHLP mRNA expression and blocks the stimulatory effect of TGF-beta on PTHLP mRNA expression. Glucocortical steroid hormone can suppress PTHLP mRNA expression and release of bioactive PTHLP in certain PTHLP-producing tumors. The regulation of PTHLP expression by female sex steroid hormones is still unclear. PTHLP is a downstream target for RAS and SRC, K-ras mutation increases PTHLP expression while a farnesyltransferase inhibitor known to inhibit RAS function can decrease PTHLP expression. The von Hippel-Lindau tumor suppressor protein also negatively regulates PTHLP expression at the post-transcriptional level.

Localisation PTHLP is a secreted polyhormone and is localized in the Golgi apparatus in the cytoplasm. However, in some cells, PTHLP can be detected in the nucleus by immunochemistry. The growth-inducing effect of NLS-containing mid-region PTHLP peptide in breast cancer is dependent on both internalization into the cytoplasm and subsequent translocation to the nucleus. PTHLP travels from the cytosol to the nucleus with the help of the nuclear transport factor importin beta1. Importin beta1 enhanced the association of PTHLP with microtubules, and the microtubule cytoskeleton plays an important role in protein transport to the nucleus. The site of recognition of PTHLP is the N-terminal half of importin, which can also bind Ran and nucleoporin, and is sufficient for PTHLP nuclear import.

Function PTHLP is a growth factor, a PTH-like calciotropic hormone, a developmental regulatory molecule, and a muscle relaxant. The diverse activities of PTHLP result not only from processing of the precursor into multiple hormones, but from use of multiple receptors. It is clear that the Type 1 Parathyroid Hormone Receptor (PTH1R), binding both PTH (1-34) and PTHLP (1-36), is the receptor mediating the function of PTHLP (1-36), and it is the only cloned receptor for PTHLP so far. PTHLP also binds to a type of receptor in some tissues that does not bind PTH. PTHLP (67-86) activates phospholipase C signaling pathways through a receptor distinct from that activated by PTHLP (1-36) in the same cells. Unlike PTH, picomolar concentrations of the PTHLP (107-111) fragment to can activate membrane-associated PKC in osteosarcoma cells. PTHLP (107-139) exerts effects through the PKC/ERK pathway. Thus, it is highly likely that the mid-region and osteostatin peptides bind other, unique receptors, but those receptors have yet to be cloned and identified. In contrast to the receptor-mediated endrocrine and paracrine action, the mid-region PTHLP peptide contains a classic bipartite nuclear localization signal (NLS) which upon entering the nuclear compartment confers 'intracrine' actions. Details of the nuclear action of PTHLP are still lacking, but overall, nuclear PTHLP appears to be mitogenic. The translation of PTHLP initiates from both the methionine-coding AUG and a leucine-coding CUGs further downstream in its signal sequence. It appeared that when translation was initiated from CUGs, PTHLP accumulated in the nucleoli, and that when translation was initiated from AUG, PTHLP localized in both the Golgi apparatus and nucleoli. Thus, nucleolar PTHLP appears to be derived from translation initiating from both AUG and CUGs. Modulation of cell adhesion by PTHLP localized in the nucleus is a normal physiological action of PTHLP, mediated by increasing integrin gene



transcription. The promotion by PTHLP in cancer growth and metastasis may be mediated by upregulating integrin alpha6beta4 expression and activating Akt. PTHLP also interacts with beta-arrestin 1B, an important component of MAPK signaling and G-protein-coupled receptor desensitization, and this interaction requires residues 122-141 of PTHLP. Therefore, beta-arrestin 1 may mediate a novel regulatory function of PTHLP in intracellular signaling. PTHLP also play a major role in development of several tissues and organs. PTHLP stimulates the proliferation of chondrocytes and suppresses their terminal differentiation. PTHLP (107-139) is a substrate for secPHEX, and osteocalcin, pyrophosphate and phosphate are inhibitors of secPHEX activity; thus PHEX activity and PTHLP are part of a complex network regulating bone mineralization. PTHLP plays a central role in the physiological regulation of bone formation, by promoting recruitment and survival of osteoblasts, and probably plays a role in the physiological regulation of bone resorption, by enhancing osteoclast formation. Signaling by fibroblast growth factor receptor 3 and PTHLP coordinate in cartilage and bone development. PTHLP is also an essential physiological regulator of adult bone mass. PTHLP aids in normal mammary gland development and lactation as well as placental transfer of calcium. Mammary gland development depends upon a complex interaction between epithelial and mesenchymal cells that requires PTHLP. The calcium sensor (CaR) regulates PTHLP production as well as transport of calcium in the lactating mammary gland. In normal animals, mammary epithelial cells secrete a lot of PTHLP, which helps to adjust maternal metabolism to meet the calcium demands of lactation. The mid-region PTHLP peptide has also been shown to control the normal maternal-to-fetal pumping of calcium across the placenta. PTHLP is secreted from smooth muscle in many organs, usually in response to stretching. PTHLP relaxes smooth muscle. Transgenic mice that express PTHLP in vascular smooth muscle have hypotension, being consistent with a vasodilating effect of PTHLP. PTHLP is highly expressed in the skin. EGF and other similar ligands can potentially activate PTHLP gene expression in the epidermis. PTHLP can inhibit hair growth and is required for tooth eruption as shown by mouse models that manipulated the PTHLP gene.

Implicated in Humoral hypercalcemia of malignancy Disease Humoral hypercalcemia of malignancy (HHM) was first described by Albright in 1941, and is a well-known complication among cancer patients. This

syndrome is commonly encountered in advanced cancer of epithelial origin, especially squamous cell carcinoma of the lung. Studies of the 'humors' secreted by cancer that causes hypercalcemia led to the discovery of 3 classes of peptides: parathyroid-like peptides, growth factor-like peptides, and bone-resorbing factors. Then protein purification led to molecular studies that cloned cDNAs for PTHLH. A study suggested that the PTHLH may be responsible for the abnormal calcium metabolism in HHM.

Prognosis The median survival after the first occurrence of hypercalcemia is 66 days in patients with serum PTHLP inferior or equal to 21 pmol/L and 33 days in patients with PTHLP superior to 21 pmol/L. In hypercalcemia of malignancy, raised serum levels of PTHLP indicate a more advanced tumor state and an extremely poor prognosis.

Autocrine promotion of tumor progression Prognosis In the absence of hypercalcemia, approximately 17% of patients with gastroesophageal carcinoma have elevated levels of PTHLP, and the increase in PTHLP was associated with a poor prognosis.

Oncogenesis mRNA for the PTH1R was detected many tumors expressing PTHLP; thus the PTHLP produced by these tumors may act in an autocrine or paracrine fashion. PTHLP (1-34) treatment resulted in an increase in proliferation in prostate cancer cells which may require androgen in some cell lines. In breast cancer cells, PTHLP regulates CDC2 and CDC25B via PTH1R in a cAMP-independent manner, and PTHLP promotes cell migration through induction of ITGA6, PAI-1, and KISS-1, and promotes proliferation through induction of KISS-1. These pieces of evidence together suggest that PTHLP and PTH1R together play an important role in the autocrine/paracrine promotion of tumor proliferation in some cancers.

Bone metastasis Disease Breast cancer

Oncogenesis PTHLP is a mediator of the bone destruction associated with osteolytic metastasis. Patients with PTHLP-expressing breast carcinoma are more likely to develop bone metastasis, and bone metastasis expresses PTHLP in more than 90% of cases as compared with less than 20% of cases of metastasis to other sites. In breast cancer, osteolytic metastases are the most common. PTHLP is a common osteolytic factor, and other osteolytic factors include vascular endothelial



growth factor and interleukin 8 and interleukin 11. Since osteoblasts are the main regulators of osteolytic osteoclasts, stimulation of osteoblasts can paradoxically increase osteoclast function. Simultaneous expression of osteoblastic and osteolytic factors can produce mixed metastases. PTHLP expression by cancer cells may provide a selective growth advantage in bone because PTHLP stimulates osteoclastic bone resorption to release growth factors such as TGF-beta from the bone matrix. TGF-beta in turn will activate by osteoclastic bone resorption and enhance PTHLP expression and tumor cell growth, thus completing a vicious cycle (See diagram). Taken together, PTHLP expression by breast carcinoma cells enhance the development and progression of breast carcinoma metastasis to bone. Alternatively, cytokines such as IL-8 initiate the process of osteoclastic bone resorption in the early stages of breast cancer metastasis, and PTHLP expression is induced to stimulate the vicious cycle of osteolysis at a later stage. Certain cancer treatments, especially sex steroid hormone deprivation therapies, stimulate bone loss. Bone resorption will result in the release of bone growth factors, which may inadvertently facilitate bone metastasis. Treatment with bisphosphonates will prevent bone resorption and reduce the release of bone growth factors.

The interactions among breast cancer cells, osteoblasts and osteoclasts define a feedback loop that promote breast cancer growth in the bone microenvironment.

Cachexia in hypercalcemia of malignancy Oncogenesis PTHLP induces a wasting/cachectic syndrome. PTHLP leads to decreased physical activity and lowered energy metabolism independently of the effects of

hypercalcemia and proinflammatory cytokines. In a rodent model, PTHLP induces a cachectic syndrome (in addition to inducing hypercalcemia of malignancy) by changing the mRNA levels of orexigenic and anorexigenic peptides, except leptin and orexin. Expression of cachexia-inducing cytokines such as interleukin-6 and leukemia inhibitory factor is increased by PTHLP. Animal data suggest that humanized antibody against PTHLP may be effective for patients with hypercalcemia and cachexia in patients with humoral hypercalcemia of malignancy.

References Albright F. Case records of the Massachusetts General Hospital (case 27461). New Eng J Med 1941;225:789-791.

Suva LJ, Winslow GA, Wettenhall RE, Hammonds RG, Moseley JM, Diefenbach-Jagger H, Rodda CP, Kemp BE, Rodriguez H, Chen EY et al. A parathyroid hormone-related protein implicated in malignant hypercalcemia: cloning and expression. Science 1987;237:893-896.

Broadus AE, Mangin M, Ikeda K, Insogna KL, Weir EC, Burtis WJ, Stewart AF. Humoral hypercalcemia of cancer. Identification of a novel parathyroid hormone-like peptide. N Engl J Med 1988;319:556-563. (Review).

Mangin M, Webb AC, Dreyer BE, Posillico JT, Ikeda K, Weir EC, Stewart AF, Bander NH, Milstone L, Barton DE et al. Identification of a cDNA encoding a parathyroid hormone-like peptide from a human tumor associated with humoral hypercalcemia of malignancy. Proc Natl Acad Sci USA 1988;85:597-601.

Ikeda K, Lu C, Weir EC, Mangin M, Broadus AE. Transcriptional regulation of the parathyroid hormone-related peptide gene by glucocorticoids and vitamin D in a human C-cell line. J Biol Chem 1989;264:15743-15746.

Yasuda T, Banville D, Hendy GN, Goltzman D. Characterization of the human parathyroid hormone-like peptide gene. Functional and evolutionary aspects. J Biol Chem 1989;264:7720-7725.

Kasono K, Isozaki O, Sato K, Sato Y, Shizume K, Ohsumi K, Demura H. Effects of glucocorticoids and calcitonin on parathyroid hormone-related protein (PTHrP) gene expression and PTHrP release in human cancer cells causing humoral hypercalcemia. Jpn J Cancer Res 1991;82:1008-1014.

Powell GJ, Southby J, Danks JA, Stillwell RG, Hayman JA, Henderson MA, Bennett RC, Martin TJ. Localization of parathyroid hormone-related protein in breast cancer metastases: increased incidence in bone compared with other sites. Cancer Res 1991;51:3059-3061.

Gagnon L, Jouishomme H, Whitfield JF, Durkin JP, MacLean S, Neugebauer W, Willick G, Rixon RH, Chakravarthy B. Protein kinase C-activating domains of parathyroid hormone-related protein. J Bone Miner Res 1993;8:497-503.

Li X, Drucker DJ. Parathyroid hormone-related peptide is a downstream target for ras and src activation. J Biol Chem 1994;269:6263-6266.

Pecherstorfer M, Schilling T, Blind E, Zimmer-Roth I, Baumgartner G, Ziegler R, Raue F. Parathyroid hormone-related protein and life expectancy in hypercalcemic cancer patients. J Clin Endocrinol Metab 1994;78:1268-1270.

Southby J, O'Keeffe LM, Martin TJ, Gillespie MT. Alternative promoter usage and mRNA splicing pathways for parathyroid hormone-related protein in normal tissues and tumours. Br J Cancer 1995;72:702-707.



Guise TA, Yin JJ, Taylor SD, Kumagai Y, Dallas M, Boyce BF, Yoneda T, Mundy GR. Evidence for a causal role of parathyroid hormone-related protein in the pathogenesis of human breast cancer-mediated osteolysis. J Clin Invest 1996;98:1544-1549.

Orloff JJ, Ganz MB, Nathanson MH, Moyer MS, Kats Y, Mitnick M, Behal A, Gasalla-Herraiz J, Isales CM. A midregion parathyroid hormone-related peptide mobilizes cytosolic calcium and stimulates formation of inositol trisphosphate in a squamous carcinoma cell line. Endocrinology 1996;137:5376-5385.

Kurebayashi J, Sonoo H. Parathyroid hormone-related protein secretion is inhibited by oestradiol and stimulated by antioestrogens in KPL-3C human breast cancer cells. Br J Cancer 1997;75:1819-1825.

Sugimoto T, Shiba E, Watanabe T, Takai S. Suppression of parathyroid hormone-related protein messenger RNA expression by medroxyprogesterone acetate in breast cancer tissues. Breast Cancer Res Treat 1999;56:11-23.

Yin JJ, Selander K, Chirgwin JM, Dallas M, Grubbs BG, Wieser R, Massagué J, Mundy GR, Guise TA. TGF-beta signaling blockade inhibits PTHrP secretion by breast cancer cells and bone metastases development. J Clin Invest 1999;103:197-206.

Guise TA. Molecular mechanisms of osteolytic bone metastases. Cancer 2000;88:2892-2898. (Review).

Asadi F, Faraj M, Malakouti S, Kukreja SC. Effect of parathyroid hormone related protein, and dihydrotestosterone on proliferation and ornithine decarboxylase mRNA in human prostate cancer cell lines. Int Urol Nephrol 2001;33:417-422.

Boileau G, Tenenhouse HS, Desgroseillers L, Crine P. Characterization of PHEX endopeptidase catalytic activity: identification of parathyroid-hormone-related peptide107-139 as a substrate and osteocalcin, PPi and phosphate as inhibitors. Biochem J 2001;355:707-713.

Clemens TL, Cormier S, Eichinger A, Endlich K, Fiaschi-Taesch N, Fischer E, Friedman PA, Karaplis AC, Massfelder T, Rossert J, Schlüter KD, Silve C, Stewart AF, Takane K, Helwig JJ. Parathyroid hormone-related protein and its receptors: nuclear functions and roles in the renal and cardiovascular systems, the placental trophoblasts and the pancreatic islets. Br J Pharmacol 2001;134:1113-1136.

de Miguel F, Fiaschi-Taesch N, López-Talavera JC, Takane KK, Massfelder T, Helwig JJ, Stewart AF. The C-terminal region of PTHrP, in addition to the nuclear localization signal, is essential for the intracrine stimulation of proliferation in vascular smooth muscle cells. Endocrinology 2001;142:4096-4105.

Deftos LJ, Burton D, Hastings RH, Terkeltaub R, Hook VY. Comparative tissue distribution of the processing enzymes 'prohormone thiol protease,' and prohormone convertases 1 and 2, in human PTHrP-producing cell lines and mammalian neuroendocrine tissues. Endocrine 2001;15:217-224.

Hook VY, Burton D, Yasothornsrikul S, Hastings RH, Deftos LJ. Proteolysis of ProPTHrP(1-141) by 'prohormone thiol protease' at multibasic residues generates PTHrP-related peptides: implications for PTHrP peptide production in lung cancer cells. Biochem Biophys Res Commun 2001;285:932-938.

Iguchi H, Onuma E, Sato K, Sato K, Ogata E. Involvement of parathyroid hormone-related protein in experimental cachexia induced by a human lung cancer-derived cell line established from a bone metastasis specimen. Int J Cancer 2001;94:24-27.

Kamai T, Arai K, Koga F, Abe H, Nakanishi K, Kambara T, Furuya N, Tsujii T, Yoshida KI. Higher expression of K-ras is associated with parathyroid hormone-related protein-induced hypercalcaemia in renal cell carcinoma. BJU Int 2001;88:960-966.

Kunakornsawat S, Rosol TJ, Capen CC, Middleton RP, Hannah SS, Inpanbutr N. Effects of 1,25(OH)2D3, EB1089, and analog V on PTHrP production, PTHrP mRNA expression and cell growth in SCC 2/88. Anticancer Res 2001;21:3355-3363.

Lindemann RK, Ballschmieter P, Nordheim A, Dittmer J. Transforming growth factor beta regulates parathyroid hormone-related protein expression in MDA-MB-231 breast cancer cells through a novel Smad/Ets synergism. J Biol Chem 2001;276:46661-46670.

Amizuka N, Oda K, Shimomura J, Maeda T. Biological action of parathyroid hormone (PTH)-related peptide (PTHrP) mediated either by the PTH/PTHrP receptor or the nucleolar translocation in chondrocytes. Anat Sci Int 2002;77:225-236.

Conlan LA, Martin TJ, Gillespie MT. The COOH-terminus of parathyroid hormone-related protein (PTHrP) interacts with beta-arrestin 1B. FEBS Lett 2002;527:71-75.

Lam MH, Thomas RJ, Loveland KL, Schilders S, Gu M, Martin TJ, Gillespie MT, Jans DA. Nuclear transport of parathyroid hormone (PTH)-related protein is dependent on microtubules. Mol Endocrinol 2002;16:390-401.

Bendre M, Gaddy D, Nicholas RW, Suva LJ. Breast cancer metastasis to bone: it is not all about PTHrP. Clin Orthop Relat Res 2003;S39-45. (Review).

Conti E. The hitchhiker's guide to the nucleus. Nat Struct Biol 2003;10:8-9.

Lindemann RK, Braig M, Hauser CA, Nordheim A, Dittmer J. Ets2 and protein kinase C epsilon are important regulators of parathyroid hormone-related protein expression in MCF-7 breast cancer cells. Biochem J 2003;372:787-797.

Sato K, Onuma E, Yocum RC, Ogata E. Treatment of malignancy-associated hypercalcemia and cachexia with humanized anti-parathyroid hormone-related protein antibody. Semin Oncol 2003;30:167-173.

Amizuka N, Davidson D, Liu H, Valverde-Franco G, Chai S, Maeda T, Ozawa H, Hammond V, Ornitz DM, Goltzman D, Henderson JE. Signalling by fibroblast growth factor receptor 3 and parathyroid hormone-related peptide coordinate cartilage and bone development. Bone 2004;34:13-25.

Bisello A, Horwitz MJ, Stewart AF. Parathyroid hormone-related protein: an essential physiological regulator of adult bone mass. Endocrinology 2004;145:3551-3553.

Cho YM, Lewis DA, Koltz PF, Richard V, Gocken TA, Rosol TJ, Konger RL, Spandau DF, Foley J. Regulation of parathyroid hormone-related protein gene expression by epidermal growth factor-family ligands in primary human keratinocytes. J Endocrinol 2004;181:179-190.

Massfelder T, Lang H, Schordan E, Lindner V, Rothhut S, Welsch S, Simon-Assmann P, Barthelmebs M, Jacqmin D, Helwig JJ. Parathyroid hormone-related protein is an essential growth factor for human clear cell renal carcinoma and a target for the von Hippel-Lindau tumor suppressor gene. Cancer Res 2004;64:180-188.

Sellers RS, Luchin AI, Richard V, Brena RM, Lima D, Rosol TJ. Alternative splicing of parathyroid hormone-related protein mRNA: expression and stability. J Mol Endocrinol 2004;33:227-241.

Dackiw A, Pan J, Xu G, Yeung SC. Modulation of parathyroid hormone-related protein levels (PTHrP) in anaplastic thyroid cancer. Surgery 2005;138:456-463.

Deans C, Wigmore S, Paterson-Brown S, Black J, Ross J, Fearon KC. Serum parathyroid hormone-related peptide is associated with systemic inflammation and adverse prognosis in gastroesophageal carcinoma. Cancer 2005;103:1810-1818.

Guise TA, Kozlow WM, Heras-Herzig A, Padalecki SS, Yin JJ, Chirgwin JM. Molecular mechanisms of breast cancer



metastases to bone. Clin Breast Cancer 2005;5 Suppl:S46-53. (Review).

Onuma E, Tsunenari T, Saito H, Sato K, Yamada-Okabe H, Ogata E. Parathyroid hormone-related protein (PTHrP) as a causative factor of cancer-associated wasting: possible involvement of PTHrP in the repression of locomotor activity in rats bearing human tumor xenografts. Int J Cancer 2005;116:471-478.

Rabbani SA, Khalili P, Arakelian A, Pizzi H, Chen G, Goltzman D. Regulation of parathyroid hormone-related peptide by estradiol: effect on tumor growth and metastasis in vitro and in vivo. Endocrinology 2005;146:2885-2894.

Ardeshirpour L, Dann P, Pollak M, Wysolmerski J, VanHouten J. The calcium-sensing receptor regulates PTHrP production and calcium transport in the lactating mammary gland. Bone 2006;38:787-793.

de Gortázar AR, Alonso V, Alvarez-Arroyo MV, Esbrit P. Transient exposure to PTHrP (107-139) exerts anabolic effects through vascular endothelial growth factor receptor 2 in human osteoblastic cells in vitro. Calcif Tissue Int 2006;79:360-369.

Dittmer A, Vetter M, Schunke D, Span PN, Sweep F, Thomssen C, Dittmer J. Parathyroid hormone-related protein regulates tumor-relevant genes in breast cancer cells. J Biol Chem 2006;281:14563-14572.

Iguchi H, Aramaki Y, Maruta S, Takiguchi S. Effects of anti-parathyroid hormone-related protein monoclonal antibody and osteoprotegerin on PTHrP-producing tumor-induced cachexia in nude mice. J Bone Miner Metab 2006;24:16-19.

Kumari R, Robertson JF, Watson SA. Nuclear targeting of a midregion PTHrP fragment is necessary for stimulating growth in breast cancer cells. Int J Cancer 2006;119:49-59.

Shen X, Falzon M. PTH-related protein upregulates integrin alpha6beta4 expression and activates Akt in breast cancer cells. Exp Cell Res 2006;312:3822-3834.

Alokail MS, Peddie MJ. Characterisation of ligand binding to the parathyroid hormone/parathyroid hormone-related peptide receptor in MCF7 breast cancer cells and SaOS-2 osteosarcoma cells. Cell Biochem Funct 2007;25:139-147.

Anderson JA, Grabowska AM, Watson SA. PTHrP increases transcriptional activity of the integrin subunit alpha5. Br J Cancer 2007;96:1394-1403.

Gessi M, Monego G, Calviello G, Lanza P, Giangaspero F, Silvestrini A, Lauriola L, Ranelletti FO. Human parathyroid hormone-related protein and human parathyroid hormone receptor type 1 are expressed in human medulloblastomas and regulate cell proliferation and apoptosis in medulloblastoma-derived cell lines. Acta Neuropathol (Berl) 2007;114:135-145.

Hamzaoui H, Rizk-Rabin M, Gordon J, Offutt C, Bertherat J, Bouizar Z. PTHrP P3 promoter activity in breast cancer cell lines: role of Ets1 and CBP (CREB binding protein). Mol Cell Endocrinol 2007;268:75-84.

Hashimoto H, Azuma Y, Kawasaki M, Fujihara H, Onuma E, Yamada-Okabe H, Takuwa Y, Ogata E, Ueta Y. Parathyroid hormone-related protein induces cachectic syndromes without directly modulating the expression of hypothalamic feeding-regulating peptides. Clin Cancer Res 2007;13:292-298.


Yeung SCJ. PTHLH (parathyroid hormone-like hormone). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):234-239.





SOCS2 (suppressor of cytokine signaling 2) Leandro Fernández-Pérez, Amilcar Flores-Morales

University of Las Palmas de GC, Faculty of Health Sciences, Molecular and Translational Endocrinology Group, c/ Dr. Pasteur s/n - Campus San Cristobal, 35016 - Las Palmas, Spain, (LFP); Department of Molecular Medicine and Surgery, Karolinska Institute, 17176 Stockholm, Sweden (AFM)


Online updated version: http://AtlasGeneticsOncology.org/Genes/SOCS2ID44123ch12q21.html DOI: 10.4267/2042/38528


Identity Hugo: SOCS2 Other names: CIS-2, Cytokine-inducible SH2 protein 2; CIS2, STAT induced STAT inhibitor-2; Cish2, STAT-induced STAT inhibitor 2; SOCS-2, suppressor of cytokine signaling 2; SSI-2, suppressor of cytokine signaling-2; SSI2; STATI2 Location: 12q21.33 Local order: By cytogenetic and radiation hybrid mapping, SOCS-2 has been mapped to chromosome 12q21.3-q23 (Yandava et al., 1999).

DNA/RNA Description 6,38 kb; 3 exons. Mouse SOCS2 gene is composed of 3 exons and 2 introns (Metcalf et al., 2000). Human SOCS-2 is a functioning gene that comprises 3 exons spanning roughly 6,38 kb of genomic DNA.

Transcription 2210 bp mRNA. 1 protein (22.2 kDa; 198 aa). Although constitutively expressed SOCS2 mRNA has been detected in several tissues and cell types, its expression is, in general, induced by stimulation with different cytokines and hormones (Rico-Bautista et al., 2006). SOCS2 promoter analysis indicates the presence of AhR and STAT5 binding sites that confer responsiveness to dioxin (Boverhof et al., 2004) and GH (Vidal et al., 2006), respectively.

Protein Description 22.2 kDa; 198 aa.

Expression SOCS mRNA and protein levels are constitutively low in unstimulated cells, but their expression is rapidly induced upon cytokine stimulation, thereby creating a negative feedback loop. Its expression is, in general, induced by stimulation with different cytokines and hormones (Rico-Bautista et al., 2006).

Localisation Intracellular, cytoplasm.

Function SOCS mechanisms of action rely on their ability to bind tyrosine phosphorylated proteins through their SH2 domains, but also to bind Elongin BC through their SOCS box domains. SOCS family proteins form part of a classical negative feedback system that regulates cytokine signal transduction (Rico-Bautista et al., 2006). SOCS2 appears to be a negative regulator in the growth hormone/IGF1 signaling pathway (Metcalf et al., 2000). SOCS2 appear to be involved in regulating protein turnover, targeting proteins for proteasome-mediated degradation (Rico-Bautista et al., 2004).

SOCS2 (suppressor of cytokine signaling 2) Fernández-Pérez L, Flores-Morales A


Diagram representing the structure of SOCS proteins. At least eight proteins belonging to the SOCS family of proteins are shown (upper panel). They are characterized by the presence of an SH2 central domain and the SOCS box domain at the C-terminus. A small domain called kinase inhibitory region (KIR), only found in SOCS1 and SOCS3, is shown as a small box at the N-terminal region. SOCS proteins can interact with phosphotyrosine phosphorylated proteins through their SH2 domain and with Elongin BC through their SOCS box domain. Other proteins containing a SOCS box domain but lacking a SH2 domain are also shown (lower panel). Adapted from Elliot and Johnston (Elliott and Johnston, 2004) with modifications.

Mutations Note: SNP: increasing the risk of type 2 diabetes.

Implicated in Diabete Note: Susceptibility to type 2 diabetes (Kato et al., 2006).

Metabolism Note: SOCS2 null mice are giants but not obese (Metcalf et al., 2000). SOCS2 deficient mice have some metabolic characteristics that can be related to the enhanced GH actions (Rico-Bautista et al., 2005).

Bone Note: Analysis of SOCS2 null mice have revealed that the absence of SOCS2 induces a reduction in the trabecular and cortical volumetric bone mineral density (Lorentzon et al., 2005). SOCS2 induces the

differentiation of C2C12 mesenchymal cells into myoblasts or osteoblasts (Ouyang et al., 2006).

Neural development Note: SOCS2 plays a critical role in neuronal development, growth, and stem cell differentiation (Turnley et al., 2002).

Cancer Note: SOCS2 has been associated with cancer such as myeloid leukaemia, pulmonary adenocarcinoma, and ovarian cancer, breast cancer, and anal cancer.

References Yandava CN, Pillari A, Drazen JM. Radiation hybrid and cytogenetic mapping of SOCS1 and SOCS2 to chromosomes 16p13 and 12q, respectively. Genomics 1999;61:108-111.

Metcalf D, Greenhalgh CJ, Viney E, Willson TA, Starr R, Nicola NA, Hilton DJ, Alexander WS. Gigantism in mice lacking suppressor of cytokine signalling-2. Nature 2000;405:1069-1073.

SOCS2 (suppressor of cytokine signaling 2) Fernández-Pérez L, Flores-Morales A


Turnley AM, Faux CH, Rietze RL, Coonan JR, Bartlett PF. Suppressor of cytokine signaling 2 regulates neuronal differentiation by inhibiting growth hormone signaling. Nat Neurosci 2002;5:1155-1162.

Boverhof DR, Tam E, Harney AS, Crawford RB, Kaminski NE, Zacharewski TR. 2,3,7,8-Tetrachlorodibenzo-p-dioxin induces suppressor of cytokine signaling 2 in murine B cells. Mol Pharmacol 2004;66:1662-1670.

Elliott J, Johnston JA. SOCS: role in inflammation, allergy and homeostasis. Trends Immunol 2004;25:434-440. (Review).

Rico-Bautista E, Negrín-Martínez C, Novoa-Mogollón J, Fernández-Perez L, Flores-Morales A. Downregulation of the growth hormone-induced Janus kinase 2/signal transducer and activator of transcription 5 signaling pathway requires an intact actin cytoskeleton. Exp Cell Res 2004;294:269-280.

Lorentzon M, Greenhalgh CJ, Mohan S, Alexander WS, Ohlsson C. Reduced bone mineral density in SOCS-2-deficient mice. Pediatr Res 2005;57:223-226.

Rico-Bautista E, Greenhalgh CJ, Tollet-Egnell P, Hilton DJ, Alexander WS, Norstedt G, Flores-Morales A. Suppressor of cytokine signaling-2 deficiency induces molecular and metabolic changes that partially overlap with growth hormone-dependent effects. Mol Endocrinol 2005;19:781-793.

Kato H, Nomura K, Osabe D, Shinohara S, Mizumori O, Katashima R, Iwasaki S, Nishimura K, Yoshino M, Kobori M, Ichiishi E, Nakamura N, Yoshikawa T, Tanahashi T, Keshavarz

P, Kunika K, Moritani M, Kudo E, Tsugawa K, Takata Y, Hamada D, Yasui N, Miyamoto T, Shiota H, Inoue H, Itakura M. Association of single-nucleotide polymorphisms in the suppressor of cytokine signaling 2 (SOCS2) gene with type 2 diabetes in the Japanese. Genomics 2006;87(4):446-458.

Ouyang X, Fujimoto M, Nakagawa R, Serada S, Tanaka T, Nomura S, Kawase I, Kishimoto T, Naka T. SOCS-2 interferes with myotube formation and potentiates osteoblast differentiation through upregulation of JunB in C2C12 cells. J Cell Physiol 2006;207(2):428-436.

Rico-Bautista E, Flores-Morales A, Fernández-Perez L. Suppressor of cytokine signaling (SOCS) 2, a protein with multiple functions. Cytokine Growth Factor Rev 2006;17:431-439. (Review).

Vidal OM, Merino R, Rico-Bautista E, Fernández-Perez L, Chia DJ, Woelfle J, Ono M, Lenhard B, Norstedt G, Rotwein P, Flores-Morales A. In vivo transcript profiling and phylogenetic analysis identifies SOCS2 as a direct STAT5b target in liver. Mol Endocrinol 2006;21(1):293-311.


Fernández-Pérez L, Flores-Morales A. SOCS2 (suppressor of cytokine signaling 2). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):240-242.

Leukaemia Section Short Communication




del(11)(p12p13) Pieter Van Vlierberghe, Jules PP Meijerink

ErasmusMC/Sophia Children’s Hospital, Pediatric Oncology/Hematology, Rotterdam, The Netherlands

Published in Atlas Database: July 2007

Online updated version: http://AtlasGeneticsOncology.org/Anomalies/del11p12p13ID1351.html DOI: 10.4267/2042/38529


Clinics and pathology Disease T-cell acute lymphoblastic leukemia (T-ALL).

Epidemiology About 5% of T-ALL patients.

Prognosis Currenlty, no relation between the cryptic deletion, del(11)(p12p13), and prognosis could be established. This could be due to the limited patient numbers in the study.

Genetics Note: The cryptic deletion, del(11)(p12p13) was identified using microarray-based comparative genome hybridisation (array-CGH). The deleted region is about 3 Mb in size and the telomeric breakpoint of these deletions is situated in or near the LMO2 oncogene. Variances in the centromeric breakpoints is detected.

Cytogenetics Variants One of the T-ALL patients showed a cryptic deletion, del(11)(p12p13), that did not target the LMO2 oncogene. Indeed, this case showed no ectopic LMO2 expression. Therefore, this genomic region could potentially contain a tumor supressor gene that also contributes to T-ALL pathogenesis.

Genes involved and Proteins RBTN2/LMO2 Location: 11p13

Protein LMO2 encodes a protein that participates in the transcription factor complex, which includes E2A, TAL1, GATA1, and LDB1 in erythroid cells. Within

this transcription complex, LMO2 mediates the protein-protein interactions by recruiting LDB1, whereas TAL1, GATA1, and E2A regulate the binding to specific DNA target sites. This complex regulates the expression of several genes in various cellular backgrounds including C-KIT, EKLF, and RALDH. In normal T-cell development, LMO2 is expressed in immature CD4/CD8 double-negative thymocytes, and is down-regulated during T-cell maturation.

Results of the chromosomal anomaly Hybrid gene Note: Ectopic expression of the LMO2 oncogene due to the removal of a negative regulatory element situated upstream of the LMO2 gene, leading to activation of the proximal LMO2 promoter. In one T-ALL case, this recurrent deletion resulted in a RAG2-LMO2 fusion gene, bringing the LMO2 gene under the control of RAG2 promoter sequences. However, it was shown that promoter substitution was not the main activational mechanism as none of the other del(11)(p12p13) positive cases showed a similar RAG2-LMO2 fusion gene. In addition, RQ-PCR analysis revealed that the expression of the RAG2-LMO2 fusion is much lower than the wildtype LMO2 expression from the proximal LMO2 gene promoter.

References Van Vlierberghe P, van Grotel M, Beverloo HB, Lee C, Helgason T, Buijs-Gladdines J, Passier M, van Wering ER, Veerman AJP, Kamps WA, Meijerink JPP, Pieters R. The cryptic chromosomal deletion del(11)(p12p13) as a new activation mechanism of LMO2 in pediatric T-cell acute lymphoblastic leukemia. Blood 2006;108(10):3520-3529.


Van Vlierberghe P, Meijerink JPP. del(11)(p12p13). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):243.





t(3;5)(q26;q34) Jean-Loup Huret

Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France


Online updated version: http://AtlasGeneticsOncology.org/Anomalies/t0305q26q34ID1278.html DOI: 10.4267/2042/38530


Identity

t(3;5)(q26;q34) G-banding – Coutesy Melanie Zenger and Claudia Haferlach.

Clinics and pathology Disease Acute myeloid leukaemia (AML)

Epidemiology Only two cases to date, a 48-year-old female patient and a male patient of unknown age, both with M2 AML.

Prognosis No data.

Cytogenetics Cytogenetics morphological Sole anomaly in both cases.

Genes involved and Proteins Note: The partner of EVI1 is yet unknown.

EVI1 Location: 3q26.2

Protein Transcrition factor; EVI1 targets include: GATA2, ZBTB16/PLZF, ZFPM2/FOG2, JNK and the PI3K/AKT pathway. Role in cell cycle progression, likely to be cell-type dependant; antiapoptotic factor; involved in neuronal development organogenesis; role in hematopoietic differentiation.

References Sendi HS, Elghezal H, Temmi H, Hichri H, Gribaa M, Elomri H, Meddeb B, Ben Othmane T, Elloumi M, Saad A,. Cytogenetic analysis in 139 Tunisian patients with de novo acute myeloid leukemia. Ann Genet 2002;45:29-32.

Poppe B, Dastugue N, Vandesompele J, Cauwelier B, De Smet B, Yigit N, De Paepe A, Cervera J, Recher C, De Mas V, Hagemeijer A, Speleman F. EVI1 is consistently expressed as principal transcript in common and rare recurrent 3q26 rearrangements. Genes Chromosomes Cancer 2006;45:349-356.

Wieser R. The oncogene and developmental regulator EVI1: expression, biochemical properties, and biological functions. Gene 2007 Jul 15;396(2):346-57.


Huret JL. t(3;5)(q26;q34). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):244.





t(3;9)(q26;p23) Jean-Loup Huret



Online updated version: http://AtlasGeneticsOncology.org/Anomalies/t0309q26p23ID1279.html DOI: 10.4267/2042/38531


Clinics and pathology Disease T-cell non Hodgkin lymphoma (T-cell NHL). Note: This is one of the very rare cases of EVI1 involvement in lymphoid malignancies.

Epidemiology Only one case to date, a 11-year-old boy.

Prognosis No data.

Cytogenetics Cytogenetics morphological Sole anomaly.



Protein Transcrition factor; EVI1 targets include: GATA2, ZBTB16 PLZF, ZFPM2/FOG2, JNK and the PI3K/AKT pathway. Role in cell cycle progression, likely to be cell-type dependant; antiapoptotic factor; involved in neuronal development organogenesis; role in hematopoietic differentiation.

References Poppe B, Dastugue N, Vandesompele J, Cauwelier B, De Smet B, Yigit N, De Paepe A, Cervera J, Recher C, De Mas V, Hagemeijer A, Speleman F. EVI1 is consistently expressed as principal transcript in common and rare recurrent 3q26 rearrangements. Genes Chromosomes Cancer 2006;45:349-356.

Wieser R. The oncogene and developmental regulator EVI1: expression, biochemical properties, and biological functions. Gene 2007 396:346-357.


Huret JL. t(3;9)(q26;p23). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):245.





t(3;17)(q26;q22) Jean-Loup Huret





Identity

t(3;17)(q26;q22) G-banding – Coutesy Melanie Zenger and Claudia Haferlach.

Clinics and pathology Disease Chronic myelogenous leukaemia with t(9;22)(q34;q11) and blast crisis of CML (BC-CML) (4 cases altogether), other myeloproliferative syndromes, Acute myeloid leukaemia (AML) in 4 cases (one M1, 2 therapy related AML,one of which after breast cancer).

Epidemiology At least 9 cases to date, aged 62 years (median, range: 49-78); sex ratio was 5M/3F.

Prognosis No data.

Cytogenetics Cytogenetics morphological Sole anomaly in 3 cases, with t(9;22)(q34;q11) in 4 cases (one of which was a complex translocation), with del(5q) (1 case), and with del(7q), and +21 in 1 case.



Protein Transcrition factor; EVI1 targets include: GATA2, ZBTB16/PLZF, ZFPM2/FOG2, JNK and the PI3K/AKT pathway. Role in cell cycle progression, likely to be cell-type dependant; antiapoptotic factor; involved in neuronal development organogenesis; role in hematopoietic differentiation.

References Mecucci C, Michaux JL, Broeckaert-Van Orshoven A, Symann M, Boogaerts M, Kulling G, Van den Berghe H. Translocation t(3;17)(q26;q22): a marker of acute disease in myeloproliferative disorders? Cancer Genet Cytogenet 1984;12:111-119.

Mugneret F, Solary E, Favre B, Caillot D, Sidaner I, Guy H. New case of t(3;17)(q26;q22) as an additional change in a Philadelphia-positive chronic myelogenous leukemia in acceleration. Cancer Genet Cytogenet 1992;60:90-92.

t(3;17)(q26;q22) Huret JL


Pedersen-Bjergaard J, Pedersen M, Roulston D, Philip P. Different genetic pathways in leukemogenesis for patients presenting with therapy-related myelodysplasia and therapy-related acute myeloid leukemia. Blood 1995;86:3542-3552.

Charrin C, Belhabri A, Treille-Ritouet D, Theuil G, Magaud JP, Fiere D, Thomas X. Structural rearrangements of chromosome 3 in 57 patients with acute myeloid leukemia: clinical, hematological and cytogenetic features. Hematol J 2002;3:21-31.

Barjesteh van Waalwijk van Doorn-Khosrovani S, Erpelinck C, van Putten WL, Valk PJ, van der Poel-van de Luytgaarde S, Hack R, Slater R, Smit EM, Beverloo HB, Verhoef G, Verdonck LF, Ossenkoppele GJ, Sonneveld P, de Greef GE, Löwenberg B, Delwel R. High EVI1 expression predicts poor survival in acute myeloid leukemia: a study of 319 de novo AML patients. Blood 2003;101:837-845.

Poppe B, Dastugue N, Vandesompele J, Cauwelier B, De Smet B, Yigit N, De Paepe A, Cervera J, Recher C, De Mas V, Hagemeijer A, Speleman F. EVI1 is consistently expressed as principal transcript in common and rare recurrent 3q26 rearrangements. Genes Chromosomes Cancer 2006;45:349-356.

Wieser R. The oncogene and developmental regulator EVI1: expression, biochemical properties, and biological functions. Gene 2007;396:346-357.


Huret JL. t(3;17)(q26;q22). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):246-247.

Leukaemia Section Mini Review




t(6;7)(q23;q34) Emmanuelle Clappier, Jean Soulier

Genome Rearrangements and Cancer Group, Hematology Laboratory and U728 INSERM, Hopital Saint-Louis and Paris 7 University, Paris, France




Identity

R-band analysis. Partial karyotype showing t(6;7)(q23;q34).

Clinics and pathology Disease T cell acute lymphoblastic leukemia (T-ALL).

Phenotype / cell stem origin T cell precursor.

Epidemiology Less than 5% among a series of non selected adult and pediatric T-ALLs (n = 3 out of 92). Six cases were described, all of them children, and 5 out of 6 being under 3 years old (1.1, 1.3, 1.8, 2.5, and 2.9 years old, respectively), which is very young for T-cell leukemia. The t(6;7) translocation could therefore be relatively common in this very low range of age.

Cytology Lymphoblasts.

Prognosis The prognosis is yet to be evaluated.

Cytogenetics Cytogenetics morphological t(6;7)(q23;q34) may be barely detectable by chromosome banding technics alone.

Left: Whole chromosome painting of chromosomes 6 (green) and 7 (red). Right: Locus-specific break-apart FISH using 6q23 probes RP11-184J4 (red) and RP11-845K5 (green) showing translocation involving the 6q23 locus.

t(6;7)(q23;q34) Clappier E, Soulier J


Cytogenetics molecular Involvement of the TCRB locus and the MYB locus can be demonstrated using flanking FISH probes.

Genes involved and Proteins TRB Location: 7q34

Protein T-cell receptor beta chain.

C-MYB Location: 6q23.3

DNA / RNA Spans over 38 kb, 15 exons (and additional alternative exons), mRNA 3.3 kb.

Protein v-myb myeloblastosis viral oncogene homolog. Transcription factor: 640 amino acids.

AHI-1 Location: 6q23.3

DNA / RNA Spans over 214 kb, 28 exons (and additional alternative exons), mRNA 5.5 kb.

Protein Jouberin (Abelson helper integration site 1 protein homolog) (AHI-1). 1196 amino acids including one SH3 domain and WD repeats.

Results of the chromosomal anomaly Hybrid gene Note: No fusion gene The t(6;7)(q23.3;q34) translocation results in juxtaposition of TRB regulatory sequences to the MYB-AHI1 locus. It results in deregulated expression of C-MYB, as demonstrated by skewed allelic expression.

Fusion protein Oncogenesis C-MYB is a transcription factor involved in hematopoiesis. In T-cell differentiation, discrete threshold levels of MYB activity regulate transition through distinct stages, suggesting that a deregulated expression could disturb the maturation process and play a role in oncogenesis. A potential role of AHI1 deregulation as a cofactor has to be evaluated. Of note, the same locus at 6q23.3 is also involved in short tandem duplications of a about 230 kb genomic region which includes the C-MYB gene (about 10% T-ALL in children and adults). This somatic abnormality can be detected by array-CGH, genomic Q-PCR or fiber-FISH, but not or hardly by standard metaphasic or interphasic FISH.

References Sinclair P, Harrison CJ, Jarosová M, Foroni L. Analysis of balanced rearrangements of chromosome 6 in acute leukemia: clustered breakpoints in q22-q23 and possible involvement of c-MYB in a new recurrent translocation, t(6;7)(q23;q32 through 36). Haematologica 2005;90(5):602-611.

Clappier E, Cuccuini W, Kalota A, Crinquette A, Cayuela JM, Dik WA, Langerak AW, Montpellier B, Nadel B, Walrafen P, Delattre O, Aurias A, Leblanc T, Dombret H, Gewirtz AM, Baruchel A, Sigaux F, Soulier J. The C-MYB locus is involved in chromosomal translocation and genomic duplications in human T-cell acute leukemia (T-ALL), the translocation defining a new T-ALL subtype in very young children. Blood 2007; 110 (4):1251-1261.

Lahortiga I, De Keersmaecker K, Van Vlierberghe P, Graux C, Cauwelier B, Lambert F, Mentens N, Beverloo HB, Pieters R, Speleman F, Odero MD, Bauters M, Froyen G, Marynen P, Vandenberghe P, Wlodarska I, Meijerink JP, Cools J. Duplication of the MYB oncogene in T cell acute lymphoblastic leukemia. Nat Genet 2007;39(5):593-595.


Clappier E, Soulier J. t(6;7)(q23;q34). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):248-249.

Solid Tumour Section Mini Review




Soft tissue tumors: Alveolar soft part sarcoma Jean-Loup Huret


Published in Atlas Database: Update -July 2007

Online updated version: http://AtlasGeneticsOncology.org/Tumors/AlveolSoftPartSID5125.html DOI: 10.4267/2042/38534

This article is an update of: Huret JL. Soft tissue tumors: Alveolar soft part sarcoma. Atlas Genet Cytogenet Oncol Haematol.2001;5(4):296-297. This work is licensed under a Creative Commons Attribution-Non-commercial-No Derivative Works 2.0 France Licence. © 2008 Atlas of Genetics and Cytogenetics in Oncology and Haematology

Identity Other names: Malignant nonchromaffin paraganglioma; Malignant organoid granular cell myoblastoma

Clinics and pathology Embryonic origin The histogenesis of this tumour is still unknown, despite immunohistochemistry studies and electron microscopy. It may have a myogenic origin, and might be a variant of rhabdomyosarcoma.

Epidemiology Rare tumour: represents less than 1% of soft tissues sarcomas of adults and 1-2% of soft tissues sarcomas in children. Occurs most often in the young adult, less frequently in children. Median age is 20 years in female patients, and 30 years in male patients. More frequently, patients are females (ratio M/F is 2/3).

Clinics Involve the muscles and soft tissues, in particular those of the lower extremities (buttocks, thighs and legs). This represents more than half cases in the adults. It may also arise in the upper extremities, in the head and neck regions, especially in the child, but it can also have extra muscular localizations, such as the female genital tract, the trunk, the mediastinum, or the retroperitoneum. Metastases are frequent. They occur mainly in lungs, bones, and brain. Symptoms at diagnosis may be pain and/or swelling. Diagnosis is often retarded.

Pathology Well circumscribed tumours with a multinodular pattern, haemorrhagic and necrotic. Microcopically, exhibits an alveolar structure, the center of the alveolar space being formed by detachment of necrotic cells, and with surronding capillaries (there is a more solid pattern in children). Cells are large, with abundant cytoplasm. Mitoses are rare. Secretory process with the formation of cytoplasmic membrane-bound crystals (PAS+, diastase resistant) can often be seen with electron microscopy, a feature of great diagnostic value (they are pathognomic). These granules contain monocarboxylate transporter 1 (MCT1) - CD147 complexes. Immunochemistry: in general, alveolar soft part sarcomas are negative for neuroendocrin and epithelial markers, and often positive for vimentin, muscle-specific actin, and desmin. The strong nuclear staining of an anti C-term TFE3 can be used for diagnosis (although cytogenetics and/or molecular genetics are the most relevant tools for diagnosis). To be noted is that a subset of renal cell carcinomas, the primary renal ASPSCR1-TFE3 tumour, share some morphological features with the alveolar soft part sarcoma (it may be a differential diagnosis); they also share a common genetic substratum.

Treatment Primary tumours: large surgical excision (a complete resection is of great importance) and radiation. Metastases: chemotherapy, with or without radiation or surgery, depending on the number of metastases.

Evolution Slow growing tumour, but highly angiogenic, which favours metastases dissemination.

Soft tissue tumors: Alveolar soft part sarcoma Huret JL


Metastases appear in more than half of the patients who presented without metastases at diagnosis (up to 70% in one study); however, there is a long disease-free interval before appearence of metastases (median 6 years) in these patients.

Prognosis Relatively indolent clinical course. In one study, overall survival of adult patients without metastases reached 87% at 5 years, but that of adult patients with metastases at diagnosis was only 20% at 5 years, with a median survival of 40 mths. Pediatric cases had a better prognosis, with a 5 years survival of 80% for all cases included, reaching 91% in cases without metastases. Median survival in patients without metastases at diagnosis was noted above 10 years in a large -but old (period 1923-1986)- study, and it may be expected that progress has been made. Due to the rarity of the disease and its long course, survival data are outdated.

Cytogenetics Cytogenetics morphological t(X;17)(p11;q25) is found in all alveolar soft part sarcomas so far studied, but also in primary renal ASPSCR1-TFE3 tumours. In the case of alveolar soft part sarcoma, the chromosome rearrangement is found in an unbalanced form, as a der(17)t(X;17)(p11;q25), in 80% of cases; the unbalanced form implicates: 1- the formation of a hybrid gene at the breakpoint, but also, 2- gain in Xp11-pter sequences, and loss of heterozygocity in 17q25-qter, with possible implications, although no clinical (including prognostic) nor pathological differences have so far been noted between balanced and unbalanced cases... but, again, the disease is rare, and cases with cytogenetic studies even rarer (about 25 cases). Note: the t(X;17)(p11;q25) in primary renal ASPSCR1-TFE3 tumours is balanced in all known cases.

Genes involved and Proteins Note: Retention of heterozygocity in the tumours of female patients (i.e. a normal maternal X and a normal paternal X are present, in addition to the Xp11-pter involved in the translocation) has been noted in all (n=7) female cases studied, showing that the translocation occurred in G2 phase.

Genes TFE3 Location: Xp11

DNA/RNA 8 exons.

Protein Transcription factor; member of the basic helix-loop-helix family (b-HLH) of transcription factors primarily found to bind to the immunoglobulin enchancer muE3 motif.

ASPSCR1 Location: 17q25

Protein Contains an UBX domain, ASPSCR1 binds SLC2A4 (solute carrier family 2 (facilitated glucose transporter), member 4, also called GLUT4) endocytosed from the plasma membrane into vesicles. SLC2A4 is retained in the cell by ASPSCR1 in the absence of insulin. Insulin stimulates the release of retained SLC2A4 to exocytosis, allowing the rapid mobilization of glucose transporters to the cell surface.

Result of the chromosomal anomaly Hybride Gene Description 5' ASPSCR1 - 3' TFE3; the reciprocal 5' TFE3 - 3' ASPSCR1 is most often absent. ASPSCR1 is fused in frame either to TFE3 exon 3 or to exon 4 (type 1 and type 2 fusions respectively).

Fusion protein Description 234 NH2 term amino acids from ASPSCR1, fused to the 280 or 315 C term amino acids from TFE3, including the activation domain, the helix-loop-helix, and the leucine zipper from TFE3.

References Lieberman PH, Brennan MF, Kimmel M, Erlandson RA, Garin-Chesa P, Flehinger BY. Alveolar soft-part sarcoma. A clinico-pathologic study of half a century. Cancer 1989;63:1-13.

Cullinane C, Thorner PS, Greenberg ML, Kwan Y, Kumar M, Squire J. Molecular genetic, cytogenetic, and immunohistochemical characterization of alveolar soft-part sarcoma. Implications for cell of origin. Cancer 1992;70(10):2444-2450.

van Echten J, van den Berg E, van Baarlen J, van Noort G, Vermey A, Dam A, Molenaar WM. An important role for chromosome 17, band q25, in the histogenesis of alveolar soft part sarcoma. Cancer Genet Cytogenet 1995;82(1):57-61.

Heimann P, Devalck C, Debusscher C, Sariban E, Vamos E. Alveolar soft-part sarcoma: further evidence by FISH for the involvement of chromosome band 17q25. Genes Chromosomes Cancer 1998;23(2):194-197.

Ordóñez NG, Mackay B. Alveolar soft-part sarcoma: a review of the pathology and histogenesis. Ultrastruct Pathol 1998;22:275-292.

Joyama S, Ueda T, Shimizu K, Kudawara I, Mano M, Funai H, Takemura K, Yoshikawa H. Chromosome rearrangement at 17q25 and xp11.2 in alveolar soft-part sarcoma: A case report and review of the literature. Cancer 1999;86:1246-1250.

Soft tissue tumors: Alveolar soft part sarcoma Huret JL


Ordóñez NG. Alveolar soft part sarcoma: a review and update. Adv Anat Pathol 1999;6:125-139.

Casanova M, Ferrari A, Bisogno G, Cecchetto G, Basso E, De Bernardi B, Indolfi P, Fossati Bellani F, Carli M. Alveolar soft part sarcoma in children and adolescents: A report from the Soft-Tissue Sarcoma Italian Cooperative Group. Ann Oncol 2000;11:1445-1449.

Lasudry J, Heimann P. Cytogenetic analysis of rare orbital tumors: further evidence for diagnostic implication. Orbit 2000;19(2):87-95.

Argani P, Antonescu CR, Illei PB, Lui MY, Timmons CF, Newbury R, Reuter VE, Garvin AJ, Perez-Atayde AR, Fletcher JA, Beckwith JB, Bridge JA, Ladanyi M. Primary renal neoplasms with the ASPL-TFE3 gene fusion of alveolar soft part sarcoma: a distinctive tumor entity previously included among renal cell carcinomas of children and adolescents. Am J Pathol 2001;159(1):179-192.

Ladanyi M, Lui MY, Antonescu CR, Krause-Boehm A, Meindl A, Argani P, Healey JH, Ueda T, Yoshikawa H, Meloni-Ehrig A, Sorensen PHB, Mertens F, Mandahl N, van den Berghe H, Sciot R, dal Cin P, Bridge J. The der(17)t(X.17)(p11;q25) of human alveolar soft part sarcoma fuses the TFE3 transcription factor gene to ASPL, a novel gene at 17q25. Oncogene 2001;20:48-57.

Portera CA Jr, Ho V, Patel SR, Hunt KK, Feig BW, Respondek PM, Yasko AW, Benjamin RS, Pollock RE, Pisters PW. Alveolar soft part sarcoma: clinical course and patterns of metastasis in 70 patients treated at a single institution. Cancer 2001;91:585-591.

Ladanyi M, Antonescu CR, Drobnjak M, Baren A, Lui MY, Golde DW, Cordon-Cardo C. The precrystalline cytoplasmic granules of alveolar soft part sarcoma contain monocarboxylate transporter 1 and CD147. Am J Pathol 2002;160(4):1215-1221.

van Ruth S, van Coevorden F, Peterse JL, Kroon BB. Alveolar soft part sarcoma. a report of 15 cases. Eur J Cancer 2002;38(10):1324-1328.

Sandberg A, Bridge J. Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors: alveolar soft part sarcoma. Cancer Genet Cytogenet 2002;136(1):1-9.

Uppal S, Aviv H, Patterson F, Cohen S, Benevenia J, Aisner S, Hameed M. Alveolar soft part sarcoma--reciprocal translocation between chromosome 17q25 and Xp11. Report of a case with metastases at presentation and review of the literature. Acta Orthop Belg 2003;69(2):182-187.

Anderson ME, Hornicek FJ, Gebhardt MC, Raskin KA, Mankin HJ. Alveolar soft part sarcoma: a rare and enigmatic entity. Clin Orthop Relat Res 2005;438:144-148.

Huang HY, Lui MY, Ladanyi M. Nonrandom cell-cycle timing of a somatic chromosomal translocation: The t(X;17) of alveolar soft-part sarcoma occurs in G2. Genes Chromosomes Cancer 2005;44(2):170-176.

Folpe AL, Deyrup AT. Alveolar soft-part sarcoma: a review and update. J Clin Pathol 2006;59(11):1127-1132.

Kayton ML, Meyers P, Wexler LH, Gerald WL, LaQuaglia MP. Clinical presentation, treatment, and outcome of alveolar soft part sarcoma in children, adolescents, and young adults. J Pediatr Surg 2006;41(1):187-193.

Tettamanzi MC, Yu C, Bogan JS, Hodsdon ME. Solution structure and backbone dynamics of an N-terminal ubiquitin-like domain in the GLUT4-regulating protein, TUG. Protein Sci 2006;15(3):498-508.

Aulmann S, Longerich T, Schirmacher P, Mechtersheimer G, Penzel R. Detection of the ASPSCR1-TFE3 gene fusion in paraffin-embedded alveolar soft part sarcomas. Histopathology 2007;50(7):881-886.

Zarrin-Khameh N, Kaye KS. Alveolar soft part sarcoma. Arch Pathol Lab Med 2007;131(3):488-491.


Huret JL. Soft tissue tumors: Alveolar soft part sarcoma. Atlas Genet Cytogenet Oncol Haematol.2008;12(3):250-252.

Solid Tumour Section Mini Review




Bone: Subungual exostosis with t(X;6)(q13;q22) Clelia Tiziana Storlazzi, Fredrik Mertens

Department of Genetics and Microbiology, University of Bari, Bari, Italy (CTS); Department of Clinical Genetics, Lund University Hospital, Lund, Sweden (FM)


Online updated version: http://AtlasGeneticsOncology.org/Tumors/SubungExosttX6ID5526.html DOI: 10.4267/2042/38535


Identity Other names: Dupuytren's exostosis

Classification Note: Benign bone-producing neoplasm of unknown cellular origin.

Clinics and pathology Disease Subungual exostosis.

Phenotype stem cell origin Unknown.

Embryonic origin Unknown.

Etiology Unknown.

Epidemiology Affects children and young adults.

Clinics Subungual exostosis usually presents as a slowly growing, painful mass localized dorsomedially in the distal phalanx, and in contrast to osteochondroma, there is usually no continuity with the underlying cortex.

Treatment Surgical excision, but local recurrences are not uncommon.

Prognosis Excellent.

Cytogenetics Cytogenetics morphological t(X;6)(q22;q13-14).

Partial G-banding karyotype showing chromosomes 6 and X in a case of subungual exostosis. The arrows indicate the breakpoints.

Cytogenetics molecular A Probe specific for COL12A1 (RP11-815E21) identified the breakpoint in 6q14.1, as it showed splitting signals on der(X) and der(6). On the same chromosomes, these signals colocalized with the signals of RP11-815E21, encompassing the COL4A5 and IRS4 genes in band Xq22.3.

Probes RP11-815E21 (COL4A5 and IRS4); RP11-1072D13 (COL12A1).

Variants The breakpoint on chromosome 6 could be centromeric to COL4A5, in an unknown location.

Bone: Subungual exostosis with t(X;6)(q13;q22) Storlazzi CT, Mertens F


FISH experiment revealing the breakpoint regions on both chromosomes 6 and X on a case of subungual exostosis.

Genes involved and Proteins COL4A5 (alpha 5 type IV collagen) Location: Xq22.3 Note: It is currently unknown whether any of these two genes is involved in the pathogenesis of subungual exostosis. DNA/RNA Genomic (chrX:107,569,810-107,827,431). Three transcript variants: isoform 1 (NM_000495), isoform 2 (NM_033380), isoform 3 (NM_03338). Protein Three proteins, respectively encoded by the isoform 1 (695 aa), isoform 2 (1691 aa), and isoform 3 (1688 aa).

COL12A1 (collagen, type XII, alpha 1) Location: 6q13 DNA/RNA Genomic (chr6:75,850,762-75,972,343). Two transcript variants, a long (NM_004370) and a short isoform (NM_080645). Protein Two proteins: 1899 amino acids (aa) and 3063 aa, respectively encoded by the short and long transcript isoforms.

Result of the chromosomal anomaly Hybride Gene Note: No detected fusion gene.

To be noted To elucidate how the transcription of these genes is affected by the translocation, further fresh or fresh frozen samples need to be studied.

References Dal Cin P, Pauwels P, Poldermans LJ, Sciot R, Van den Berghe H. Clonal chromosome abnormalities in a so-called Dupuytren's subungual exostosis. Genes Chromosomes Cancer 1999;24:162-164.

Murphey MD, Choi JJ, Kransdorf MJ, Flemming DJ, Gannon FH. Imaging of osteochondroma: variants and complications with radiologic-pathologic correlation. Radiographics 2000;20:1407-1434. (Review).

Ilyas W, Geskin L, Joseph AK, Seraly MP. Subungual exostosis of the third toe. J Am Acad Dermatol 2001;45:S200-S201.

Zambrano E, Nosé V, Perez-Atayde AR, Gebhardt M, Hresko MT, Kleinman P, Richkind KE, Kozakewich HP. Distinct chromosomal rearrangements in subungual (Dupuytren) exostosis and bizarre parosteal osteochondromatous proliferation (Nora lesion). Am J Surg Pathol 2004;28:1033-1039.

Storlazzi CT, Wozniak A, Panagopoulos I, Sciot R, Mandahl N, Mertens F, Debiec-Rychter M. Rearrangement of the COL12A1 and COL4A5 genes in subungual exostosis: molecular cytogenetic delineation of the tumor-specific translocation t(X;6)(q13-14;q22). Int J Cancer 2006;118:1972-1976.


Storlazzi CT, Mertens F. Bone: Subungual exostosis with t(X;6)(q13;q22). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):253-254.

Cancer Prone Disease Section Mini Review




Glomuvenous malformation (GVM) Virginie Aerts, Pascal Brouillard, Laurence M Boon, Miikka Vikkula

Human Molecular Genetics (GEHU) de Duve Institute, Universite catholique de Louvain, Avenue Hippocrate 74(+5), bp. 75.39, B-1200 Brussels, Belgium


Online updated version: http://AtlasGeneticsOncology.org/Kprones/GlomuvenousID10120.html DOI: 10.4267/2042/38536


Identity Other names: Venous malformation with glomus cells (VMGLOM); Glomangioma; Multiple glomus tumor. Note: Glomuvenous malformation (GVM) is a localized bluish-purple cutaneous vascular lesion, histologically consisting of distended venous channels with flattened endothelium surrounded by variable number of maldifferentiated smooth muscle-like “glomus cells” in the wall. GVM account for 5% of venous anomalies referred to centers for vascular anomalies. Previously, these lesions have been called “multiple glomus tumors” or “glomangioma”. Inheritance: GVM is often, if not always, hereditary (64%), and transmitted as an autosomal dominant disorder. Expressivity varies, as does penetrance, which is age dependent and maximal (93%) by 20 years of age.

Clinics Phenotype and clinics There is a wide phenotypic variation between GVM patients, even within the same family (with the same germline mutation). An individual can have an extensive lesion, affecting for example a whole extremity or most of the trunk, while others have minor, scattered papulonodular lesions of a few millimetres in diameter. The lesions are often multiple, and they can affect any body part. Seven features characterize GVM lesions : (1) Colour: GVMs can be pink in infants, the most are bluish-purple; (2) Affected tissues: the lesions are localized to the skin and subcutis, and they are rarely mucosal and never extend deeply into muscles;

(3) Localization: lesions are more often located on the extremities, although they can be found all over the body; (4) Appearance: lesions are usually nodular and multifocal, raised with a cobblestone-like appearance, except for the rare plaque-like variant. They are often hyperkeratotic; (5) The lesions are not compressible; (6) The lesions are painful on palpation; (7) New lesions can appear with time, likely after trauma.

Examples of GVMs: (A) Extended GVM on leg. (B) Small GVM on knee.

Glomuvenous malformation (GVM) Aerts V, et al.


At the histological level, the mural glomus cells are positive for smooth muscle alpha-actin and vimentin, but negative for desmin, Von Willebrand factor and S-100. Under electron microscopy, glomus cells show smooth muscle myofibrils and “dense bodies”, characteristics of vascular smooth muscle cells (vSMCs). Thus, these cells are most likely incompletely or improperly differentiated vSMCs.

Neoplastic risk GVM has no neoplastic histological characteristics and never becomes malignant.

Treatment The gold-standard treatment for GVM consists of surgical resection, as lesions are superficial and rarely affect deeply the underlying muscle, and sometimes sclerotherapy. In contrast to venous malformations, the use of elastic compressive garments often aggravate pain and should thus be avoided.

Evolution GVM is a developmental lesion that grows proportionally with the child. After partial resection, recurrence is frequent. New small lesions can appear with time. The red plaque-like lesions of the young darken with age.

Cytogenetics No cytogenetic abnormally has been reported for GVM.

Genes involved and Proteins Glomulin Location: 1p22.1

DNA/RNA Description: The glomulin gene spans about 55 kbp and contains 19 exons coding for 1785 bp. Transcription: 2 kb transcript.

Protein Glomulin was identified by reverse genetics, and its function is currently unknown. Description: Glomulin gene encodes a protein of 594 amino acids (68 kDa). Expression: The high level of glomulin expression in the murine vasculature indicates that glomulin may have an important role in blood vessel development and/or maintenance.

Localisation: Glomulin is likely an intracellular protein. Function: The exact function of glomulin is unknown. Glomulin has been described to interact with FKBP12, an immunophilin that binds the immunosuppressive drugs FK506 and rapamycin. FKBP12 interacts with the TGFbeta type I receptor, and prevents its phosphorylation. Thus, FKBP12 safeguards against the ligand-independent activation of this pathway. Glomulin, through its interaction with FKBP12, could act as a repressor of this inhibition. Glomulin has also been described to interact with c-MET. Glomulin interacts with the inactive, non phosphorylated form of c-MET. When c-MET is activated by HGF, glomulin is released in a phosphorylated form. This leads to p70 S6 protein kinase (p70S6K) phosphorylation. It is not known whether glomulin activates p70S6K directly or indirectly. The p70S6K is a key regulator of protein synthesis. Glomulin could thereby control cellular events such as migration and cell division. The third reported glomulin partner is Cul7. This places glomulin in an SCF-like complex, which is implicated in protein ubiquitination and degradation.

Mutations There is no phenotype-genotype correlation in GVM. Germinal: To date, 29 different inherited mutations (deletions, insertions and nonsense substitutions) have been identified. The most 5' mutation are located in the first coding exon. The majority of them cause premature truncation of the protein and likely result in loss-of-function. One mutation deletes 3 nucleotides resulting in the deletion of an asparagine at position 394 of the protein. More than 70% of GVMs are caused by eight different mutations in glomulin: 157delAAGAA (40,7%), 108C to A (9,3%), 1179delCAA (8,1%), 421insT and 738insT (4,65% each), 554delA+556delCCT (3,5%), 107insG and IVS5-1(G to A) (2,3% each). Somatic: The phenotypic variability observed in GVM could be explained by the need of a somatic second-hit mutation. Such a mechanism was discovered in one GVM (somatic mutation 980delCAGAA), suggesting that the lesion is due to a complete localized loss-of-function of glomulin. This concept can explain why some patients have bigger lesions than others, why new lesions appear, and why they are multifocal. This could also explain, why some mutation carriers are unaffected.

Glomuvenous malformation (GVM) Aerts V, et al.


Schematic representation of glomulin : The two stars indicate the start and the stop codons, in exon 2 and 19 respectively. All known mutations are shown. Somatic second hit is in blue.

References Goodman TF, Abele DC. Multiple glomus tumors. A clinical and electron microscopic study. Arch Dermatol 1971;103(1):11-23.

Chambraud B, Radanyi C, Camonis JH, Shazand K, Rajkowski K, Baulieu EE. FAP48, a new protein that forms specific complexes with both immunophilins FKBP59 and FKBP12. Prevention by the immunosuppressant drugs FK506 and rapamycin. J biol Chem 1996;271(51):32923-32929.

Chen YG, Liu F, Massagué J. TGFbeta receptor inhibition by FKBP12. EMBO J 1997;16(13):3866-3876.

Boon LM, Brouillard P, Irrthum A, Karttunen L, Warman ML, Rudolph R, Mulliken JB, Olsen BR, Vikkula M. A gene for inherited cutaneous venous anomalies ('glomangiomas') localizes to chromosome 1p21-22. Am J Hum Genet 1999;65(1):125-133.

Brouillard P, Olsen BR, Vikkula M. High-resolution physical and transcript map of the locus for venous malformations with glomus cells (VMGLOM) on chromosome 1p21-p22. Genomics 2000;67(1):96-101.

Grisendi S, Chambraud B, Gout I, Comoglio PM, Crepaldi T. Ligand-regulated binding of FAP68 to the hepatocyte growth factor receptor. J Biol Chem 2001;276(49):46632-46638.

Irrthum A, Brouillard P, Enjolras O, Gibbs NF, Eichenfield LF, Olsen BR, Mulliken JB, Boon LM, Vikkula M. Linkage disequilibrium narrows locus for venous malformation with glomus cells (VMGLOM) to a single 1.48 Mbp YAC. Eur J Hum Genet 2001;9(1):34-38.

Brouillard P, Boon LM, Mulliken JB, Enjolras O, Ghassibé M, Matthew L, Warman O, Tan T, Olsen BR, Vikkula M. Mutations in a novel factor, Glomulin, are responsible for glomuvenous malformations ('Glomangiomas'). Am J Hum Genet 2002;70:866-874.

Arai T, Kasper JS, Skaar JR, Ali SH, Takahashi C, DeCaprio JA. Targeted disruption of P185/Cul7 gene results in abnormal vascular morphogenesis. Proc Natl Acad Sci USA 2003;100(17):9855-9860.

Boon LM, Mulliken JB, Enjolras O, Vikkula M. Glomuvenous malformations (glomangioma) and Venous malformations, Distinct clinicopathologic and genetic entities. Arch Dermatol 2004;140:971-976.

McIntyre BA, Brouillard P, Aerts V, Gutierrez-Roelens I, Vikkula M. Glomulin is predominantly expressed in vascular smooth muscle cells in the embryonic and adult mouse. Gene Expr Patterns 2004;4(3):351-358.

Brouillard P, Ghassibe M, Penington A, Boon LM, Dompmartin a, Temple IK, Cordisco M, Adams D, Piette F, Harper JI, Syed S, Boralevi F, Taieb A, Danda S, Baselga E, Enjolras O, Mulliken JB, Vikkula M. Four common glomulin mutation cause two thirds of glomuvenous malformations ('familial glomangiomas') : evidence for a founder effect. J Med Genet 2005;42(2):e13.

Boon LM, Vanwijck R. Medical and surgical treatment of venous malformations. Ann Chir Plast Esthet 2006;51(4-5):403-411.

Mallory SB, Enjolras O, Boon LM, Rogers E, Berk DR, Blei F, Baselga E, Ros AM, Vikkula M. Congenital plaque-type glomuvenous malformations presenting in childhood. Arch Dermatol 2006;142(7):892-896.

Brouillard P, Enjolras O, Boon LM, Vikkula M. GLMN and Glomuvenous Malformation. Inborn Errors of Development 2e, edited by Charles Epstein, Robert Erickson and Anthony Wynshaw-Boris, Oxford University Press, Inc.


Aerts V, Brouillard P, Boon LM, Vikkula M. Glomuvenous malformation (GVM). Atlas Genet Cytogenet Oncol Haematol.2008;12(3):255-257.

Case Report Section Paper co-edited with the European LeukemiaNet




Translocation t(1;6)(p35;p25) in B-cell lymphoproliferative disorder with evolution to Diffuse Large B-cell Lymphoma Elvira D Rodrigues Pereira Velloso, Cristina Ratis, Sérgio AB Brasil, João Carlos Guerra, Nydia S Bacal, Cristóvão LP Mangueira

Clinical Laboratory, Hospital Israelita Albert Einstein, São Paulo, Brazil (EDRPV, CAR, NSB, CLPM); Centro de Hematologia São Paulo, São Paulo, Brazil (SABB, JCG)


Online updated version: http://AtlasGeneticsOncology.org/Reports/0106RodriguesID100030.html DOI: 10.4267/2042/38537


Clinics Age and sex: 75 years old female patient. Organomegaly: - hepatomegaly; - splenomegaly; - enlarged lymph nodes; - no central nervous system involvement. Previous history: - B-cell Lymphoproliferative disorder for 8 years; - A 75-years-old female with an 8 years diagnosis of mature B-cell proliferation disorder. At August, 1999 a CBC showed high WBC, and physical examination showed enlarged lymph nodes (cervical and axillaries). Peripheral blood revealed Hb: 14,8g/dl, WBC: 21,8 x 109/l, lymphocytes 16 x 109/l, platelets 241 x 109/l and bone marrow trephine showed interstitial infiltration with small lymphocytes consistent with CLL stage A (Binet). In April, 2000 there was a significant increase in the lymph nodes and night sweats. PB immunophenotyping study showed a CD19/CD5 positive population consisted with B-CLL (Matutes' score 4). The patient was treated with 6 cycles of COP, with evolution to pulmonary nodules and axillae bulky in 2001. From 2001 to 2004, a few cycles of Chlorambucil and 7 cycles of R-COP produced a good response. In April, 2005, the PB morphology and immunophenotype were consistent to atypical B-CLL (CD19, CD20, CD23, CD25, HLA-DR, IgM, IgD, CD79b, CD38, and sKappa,positive and CD5 negative), and PB karyotype showed no clonal abnormalities in 20 metaphases. In August, 2005, there was an increased in the number of lymph nodes and

Rituximab and Fludarabine was started. The PB counts showed: Hb: 13g/dl, WBC: 69,6 x 109/l, lymphocytes: 62 x 109/l, platelets: 145 x 109/l. Inguinal lymph node biopsy showed diffuse large B-cell Lymphoma, Ki-67: 70%, cyclin D1 -, CD20 +, BCL2 +. From December, 2005 to April, 2006, 6 cycles of R-CHOP showed no response. From October, 2006 to May, 2007, regression of lymph nodes and clinical improvement was done with 6 cycles of MiCEP. At this time, cytogenetics and immunophenotyping studies of bone marrow were performed.

Blood WBC: 5,4 x 109/l; Hb: 13,6 g/dl; platelets: 169 x 109/l; blasts: 3,35 x 109/l (lymphoid cells)%. Bone marrow: 28% of lymphoid mature cells.

Cytopathology classification Cytology: B-cell Lymphoproliferative disorder (Atypical CLL) with evolution to diffuse large B-cell Lymphoma. Atypical CLL. Immunophenotype: 25% of total bone marrow cells are positive : CD20++, CD22+, CD25+, CD38, CD79b++, HLA-DR, sIgM, sIgD e sKappa ++. Rearranged Ig or Tcr: not done. Pathology: Inguinal Lymph node biopsy (August, 2005): Diffuse large B-cell Lymphoma, Ki-67: 70%, ciclina D1 -, CD20 +, BCL2 +. Electron microscopy: not done. Precise diagnosis: B-cell Lymphoproliferative disorder (Atypical CLL) with evolution to diffuse large B-cell Lymphoma.

Translocation t(1;6)(p35;p25) in B-cell lymphoproliferative disorder Rodrigues Pereira Velloso ED, et al. with evolution to Diffuse Large B-cell Lymphoma


Survival Date of diagnosis: 07-2007. Treatment: wide previous history (long previous history of chemotherapy), no chemotherapy after July, 2007. Complete remission: not applied. Treatment related death: - Relapse: - Status: Alive 09-2007. Survival: 2 months.

Karyotype Sample: bone marrow cells; culture time: 72 hours with and without TPA (o-tetradecanoyl phorbol-13-acetate); banding: G; results: 47, XX, t(1;6)(p35;p25), +12[13]/46,XX[7]. Karyotype at relapse: not done. Other molecular cytogenetic techniques: not done.

Other molecular studies Technics: not done

Partial karyotype, G-band.

Comments In 2005, the Belgian group described the t(1;6)(p35.3;p25.2) in 8 patients with unmutated B-CLL. As in this case, this rare cytogenetic entity has been described in typical or atypical CLL (8/8 cases), with evolution to diffuse large B-cell Lymphoma (3/8 cases); trisomy 12 been a common additional abnormality (3/8 cases).

References Michaux L, Wlodarska I, Rack K, Stul M, Criel A, Maerevoet M, Marichal S, Demuynck H, Mineur P, Kargar Samani K, Van Hoof A, Ferrant A, Marynen P, Hagemeijer A. Translocation t(1;6)(p35.3;p25.2): a new recurrent aberration in 'unmutated' B-CLL. Leukemia 2005;19:77-82.


Rodrigues Pereira Velloso ED, Ratis CA, Brasil SAB, Guerra JC, Bacal NS, Mangueira LM Pitangueira CP. Translocation t(1;6)(p35;p25) in B-cell lymphoproliferative disorder with evolution to Diffuse Large B-cell Lymphoma. Atlas Genet Cytogenet Oncol Haematol.2008;12(3):258-259.

Educational Item Section Oral presentation at the 6th European Cytogenetic Conference (ECC), Istanbul, July 2007, organized by the European Cytogeneticists Association.




How human chromosome aberrations are formed Albert Schinzel

Institute of Medical Genetics, Schorenstr. 16, CH-8603 Schwerzenbach, Switzerland


Online updated version: http://AtlasGeneticsOncology.org/Educ/ChromAberFormedID30065ES.html DOI: 10.4267/2042/38538


I- Introduction II- Origin and mechanisms of formation of chromosome aberrations III- Chromosome aberrations, classification IV- Modes of determination of the mechanisms of formation of chromosome aberrations V- Microsatellite marker analysis VI- Summary of parental origin of chromosome aberrations VII- Origin of Ullrich-Turner syndrome 45,X VIII- Origin of recurrent free trisomy 21 IX- Interchromosal effect (ICE) X- Origin of mosaic trisomy XI- Origin of interstitial (micro-)deletions, interchromosomal versus intrachromosomal XII- Frequent interstitial microdeletions (15q12, 7q11.23, 22q11.2) XIII- Origin of mosaic duplications (de novo) XIV- Origin of additional isochromosomes and isodicentric chromosomes XV- Chaotic chromosome aberrations XVI- Origin of multipe structural chromosome aberrations XVII- Primary and secondary chromosome aberrations XVIII- Conclusion

I- Introduction Characteristics of the species homo sapiens: - Many! - Among others: excessively high incidence of

reproductive failure and chromosome aberrations. - Determination of origin and mechanisms of

formation of chromosome aberrations: Each newly developed technique, from Q banding over FISH and microsatellite marker analysis to CGH, has brought additional information as to the origin of chromosomal imbalance in man.

II- Origin and mechanisms of formation of chromosome aberrations Origin may be: - maternal - paternal - combined

Formation: - Nondisjunction: meiotic, pre-meiotic, post-meiotic - Rearrangement: meiotic, pre-meiotic, post-meiotic

Any combination: Incorporation of 2 sperms or of a polar body into the oocyte.

III- Chromosome aberrations, classification Numerical aberrations: - Monosomy (X/Y) - Trisomy - Sex chromosome aneuploidy - Double/triple aneuploidy - Uniparental disomy

Ploidy aberrations: - Haploidy - Triploidy

How human chromosome aberrations are formed Schinzel A


- Tetraploidy

Structural aberrations: - Deletions - Rings - Duplications - Balanced rearrangements - Combined duplication-deletion - Complex rearrangements

Mosaic and chimeras Combinations: - Numerical and structural - Numerical and ploidy, etc...

IV- Modes of determination of the mechanisms of formation of chromosome aberrations 1. Aberration per se:

- free trisomy: nondisjunction - mosaicism: either postzygotic origin or two steps - triploidy

2. Cytogenetic markers.

3. Molecular marker analysis in proband and parents.

4. Molecular marker analysis in grandparents of proband.

5. CGH.

Legend: Paternal (P) and maternal (M) chromosomes 14, the free 14 and the 14/21 translocation from the Down's offspring, Q-banded. The free 14 is of paternal origin, therefore the 14/21 is of maternal origin (from Chamberlin 1980; Hum Genet 53: 343).



V- Microsatellite marker analysis - Almost always able to determine the origin of

deletions. - Often not successful for duplications, especially

direct or inverted duplications stemming from chromatid interchanges (no third allele, often no clear intensity differences).

VI- Summary of parental origin of chromosome aberrations - Numerical, autosomes: predominantly mat. - Numerical, X chromosomes: idem. - Numerical, X and Y: overwhelming paternal origin.

- Structural, terminal deletions and rings: predominantly pat.

- Structural, extra rearranged chromosomes isochromosomes, inv dup chromosomes: predominantly mat from initial nondisjunction.

- Structural , intrachromosomal rearrangements: equal distribution.

- Structural, interchromosomal rearrangements: idem. - Uniparental disomy:

o Heterodisomy: predominantly mat from initial trisomy.

o Isodisomy: predominantly pat. - Mosaics: mostly starting with maternal trisomy

o Triploidy: o predominantly (80%) mat, incorporation

of a polar body into the oocyte; o rarer (20%) fertilization of the oocyte by

2 different sperms.

VII- Origin of Ullrich-Turner syndrome 45,X Xg studies: predominant maternal origin of the remaining X-chromosome. Expected distribution (as 45,Y is none-viable) if mat = pat: 66 vs 33%. Distribution found: 80 vs 20% (statistically significant). Parental Xg information about 306 females with 45,X Ullrich-Turner syndrome (Sanger et al., 1971).

Xg groups of Source of normal X

Number Father Mother T

+ + + unknown 150 + + - maternal 31 + - + paternal 5 + - - maternal 10 - + + maternal 60 - + - unknown 35 - - - unknown 15

Total 306 + = Xg(a+); - = Xg(a-)

VIII- Origin of recurrent free trisomy 21 Results of molecular marker studies: - 1. In siblings:

- 60% by chance - 40% parental gonadal mosaicism

- 2. In more remote relatives: - 100% by chance



IX- Interchromosal effect (ICE) - Definition: A balanced chromosome aberration increases the risk of non-disjunction for other chromosomes.

- Consequence: Prenatal cytogenetic diagnosis is indicated if one parent carries a balanced rearrangement even if unbalanced segregation cannot lead to viable offspring.

- Evidence for ICE: More familial balanced translocations found in Down syndrome patients than expected by chance.

- Evidence against an ICE: In haploid sperms of male carriers of balanced translocations there is no increase of disomies over controls.

Number

of families

Origin of the supernumerary 21 mat pat

mat. rearrangement 2 2 0

pat. rearrangement 11 11 0

X- Origin of mosaic trisomy - Mostly first trisomy: secondary somatic loss of the

third homologue. - Not infrequently: mosaicism between (maternal)

trisomy and (maternal) uniparental disomy.

XI- Origin of interstitial (micro-)deletions, interchromosomal versus intrachromosomal Principle Investigation of grandparents of the side of origin with markers flanking the deleted segment.

Williams-Beuren syndrome: - Deletion of 7q11.22 including the Elastin locus. - Supravalvular aortic stenosis. - Peripheral pulmonary stenosis. - Growth retardation. - Moderate mental retardation. - Outgoing pleasant personality. - Full lips, cheeks and lids. - Deep voice

Result:

- Switch from grandpaternal to grandmaternal origin on either side: - interchromosomal rearrangement. - meiotic origin. - low recurrence risk.

- No switch, markers on either side from grandparent: - intrachromosomal rearrangement (between 2

chromatids). - meiotic or pre- or post-meiotic origin. - not necessarily low recurrence risk.



Representative examples of microsatellite analysis at 7q11.2 carried out. The deleted region of chromosome 7 is indicated with a black bar beside the chromosome 7 ideogram. Marker D7S1870, located within the deleted region, illustrates the maternal origin of the deletion. Grandparental origin of the regions flanking the deletion are shown with markers D7S672 (proximal region) and D7S524 (distal region). XII- Frequent interstitial microdeletions (15q12, 7q11.23, 22q11.2) - Reason for their high incidence: similar short tandem

repeats. - Frequent paracentric inversions of this segment. - Tend to pair at meiosis. - Cutting out of the segment forming an inversion

loop.

XIII- Origin of mosaic duplications (de novo) Not infrequently: - First trisomy; - Second rearrangement; - Third uniparental disomy.



XIV- Origin of additional isochromosomes and isodicentric chromosomes Molecular marker analysis: - Postmeiotic: one normal, one strong allele.

- Meiotic: M1: proximal heterozygosity / M2 : vice versa.

- Results: mostly M2 maternal.

Mechanism: - first meiotic nondisjunction, - second isochromosome formation. Examples: i(8p), i(9p), i(12p), i(18p).

XV- Chaotic chromosome aberrations - Found especially at investigation of early

spontaneous abortions. - Multiple deletions, combined deletions and

duplications, etc...



- Complex balanced and unbalanced aberrations often following irradiation.

XVI- Origin of multipe structural chromosome aberrations CGH re-investigations of visible structural chromosome aberrations not infrequently detect further submicroscopic imbalances, mostly small deletions, rarer duplications. These point towards a much more complex mechanism of origin of structural aberrations than seen on the first glance and parallels the complex origin of mosaics, especially for structural and combined numerical - structural chromosome aberrations.

XVII- Primary and secondary chromosome aberrations Secondary aberrations may enable survival of an otherwise lethal unbalanced product. Examples: - Additional isochromosomes deriving from a

trisomy. - Correction of trisomy through uniparental disomy. - Secondary structural aberrations with loss of a

chromosomal segment following a trisomy. - Reduction of a complex rearrangement with multiple

breaks to a simpler one through recombination - balanced and unbalanced.

XVIII- Conclusion A distinct feature of homo sapiens is the excessively high incidence of unbalanced chromosome aberrations, especially trisomy and triploidy. Nature has an incredible phantasy and many different mechanisms to correct such unbalanced aberrations. This may happen because of a high proneness to early postzygotic numerical and structural aberrations combined with a high selection pressure. It is unknown whether primary aberrations may lead with preference to secondary imbalance. Anyway, these visible aberrations constitute the tip of an iceberg, and under the water surface are the many spontaneous miscarriages due to chromosomal imbalance.

Acknowledgements IMG Zurich:

Alessandra Baumer and Collaborators. Mariluce Riegel and Collaborators.

Europe: Collegues from many countries, especially Turkey (Seher Basaran), Poland, Hungary, Ukraine, Spain, and the ECARUCA project .


Schinzel A. How human chromosome aberrations are formed. Atlas Genet Cytogenet Oncol Haematol.2008;12(3):260-266.



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