CURRICULUM VITAE DONALD L. DURDEN, M.D., Ph.D.jacobsschool.ucsd.edu/uploads/agile_faculty/cv/durden_donald.pdfCurriculum vitae Donald L. Durden, M.D., Ph.D 04/28/16 6 2012-present

4/28/2016

CURRICULUM VITAE

DONALD L. DURDEN, M.D., Ph.D.

Professor of Pediatrics UCSD School of Medicine Rady Children’s Hospital Vice Chair for Research Department of Pediatrics

Director Pediatric Oncology Research Associate Director, Pediatric Oncology

UCSD Moores Cancer Center Email: [email protected]

Home Address: 12545 El Camino Real, Unit C San Diego, CA 92130 (678) 595-7362 (M) A. EDUCATION 7/1988-11/1992 Fellowship, Pediatric Hematology/Oncology Children’s Hospital Medical Center

Fred Hutchinson Cancer Research Center Seattle, Washington

7/1989-11/1992 Postdoctoral fellowship


7/1987-4/1988 Pediatrics Residency (year 3)

Children's Hospital and Medical Center Seattle, Washington

7/1986-6/1987 Pediatrics Residency (year 2)

St. Christopher's Hospital for Children Philadelphia, Pennsylvania

7/1985-6/1986 Internship, Department of Pediatrics

St. Christopher's Hospital for Children Philadelphia, Pennsylvania

9/1980-5/1985 Medical School (MD degree conferred 5/1985) University of Miami School of Medicine Miami, Florida

Curriculum vitae Donald L. Durden, M.D., Ph.D 04/28/16

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9/1980-5/1985 Graduate School Ph.D. (PhD degree conferred 5/1983)

Dept. of Microbiology and Immunology University of Miami School of Medicine Miami, Florida

9/1971-7/1977 B.S. (BS degree conferred 1977)

University of South Florida Tampa, Florida Major: Microbiology and Zoology

Medical licensure & Board Certifications: Indiana State #01051141 Washington State #0025156

California State #G075994 Georgia State #54999 ABP, Pediatrics (1993) ABP, Pediatric Hematology-Oncology

B. HONORS AND AWARDS 1979 Sigma Xi 1988 American Cancer Society Postdoctoral Fellowship 1989 National Research Service Award, National Institutes of Health 1997 Society for Pediatric Research (SPR) 1997 Stop Cancer Research Career Development Award 2003 Georgia Cancer Coalition Distinguished Cancer Scholar Award 2004 Aflac Endowed Professor, Emory University School of Medicine 2007 Goldhirsh Foundation Award 2009 Vice Chair for Research, Department of Pediatrics, UCSD 2012 Cancer Working Group, USWNR 2015 Editorial Board, Molecular Cancer Research (MCR)(AACR) C. FELLOWSHIPS 7/1988-11/1992 Fellowship, Pediatric Hematology/Oncology Children’s Hospital Medical Center


7/1989-11/1992 Postdoctoral fellowship


"Tyrosine phosphorylation and myeloid signal transduction" Jonathan A. Cooper, PhD, advisor

D. ACADEMIC APPOINTMENTS


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10/2009-present Professor, Pediatrics

UCSD School of Medicine Vice Chair for Research Founder and Director Rady UCSD Biorepository (RUB) Director Pediatric Oncology Research Associate Director, Pediatric Oncology UCSD Moores Cancer Center La Jolla, CA, 92093

12/2003-10/2009 Endowed Professor, Pediatrics Scientific Director, Aflac Cancer Center and Blood Disorders Services Children’s Healthcare of Atlanta Emory University School of Medicine Atlanta, GA 5/1999-12/2003 Associate Professor, Pediatrics

Biochemistry and Molecular Biology Herman B Wells Center for Pediatric Research James Whitcomb Riley Hospital for Children Indiana University Medical Center Cancer Research Institute Indianapolis, Indiana

5/1999-12/2003 Member, Indiana University Cancer Center IU School of Medicine Indianapolis, Indiana 11/1992-4/1999 Assistant Professor of Pediatrics

USC School of Medicine Division of Hematology/Oncology Children’s Hospital Los Angeles Los Angeles, California

1/1993-4/1999 Member, Norris Cancer Center

USC School of Medicine Los Angeles, California

E. PROFESSIONAL BACKGROUND 10/2015-present Founding Member, Rady Pediatric Genomics and Systems Biology Institute at Rady

Children’s Hospital, Directory Rady UCSD Biorepository (RUB) 10/2015-present Member, Center for Microbiome Innovation UCSD Schools of Medicine and Bioengineering 12/2009-present Member, Drug Discovery Institute (ORU) UCSD School of Medicine


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12/2009-present Member, Senior Leadership Executive Committee Associate Director, Pediatric Oncology Moores UCSD Cancer Center UCSD School of Medicine 12/2009-present Attending Physician Hematology-Oncology Program UCSD/Rady Children’s Hospital San Diego, CA 12/2003-10/2009 Member, Winship Cancer Center Emory University School of Medicine Atlanta, GA 12/2003-10/2009 Attending Physician Hematology-Oncology Program Aflac Cancer Center Children’s Healthcare of Atlanta (CHOA) Emory University School of Medicine Atlanta, GA 1/2007 Pediatric Executive Leadership Training Program (PEP) 12/2003-10/2009 Member, Drug Discovery Program Department of Pharmacology WCI-Emory University School of Medicine Atlanta, GA 12/2003-10/2009 Member, Brain Tumor Research Program Member, Prostate Cancer Research Program Winship Cancer Institute (WCI) Emory University School of Medicine Atlanta, GA 8/2002-12/2003 Director, Pediatric Oncology Research Group Indiana University School of Medicine Indiana University Cancer Center Indianapolis, Indiana 5/2000-12/2003 Member, Indiana University Center for Vascular Biology Indiana University School of Medicine Indianapolis, Indiana 5/1999-12/2003 Attending Physician

Stem Cell Transplantation Program Riley Children’s Hospital Indiana University School of Medicine Indianapolis, Indiana

5/1999-12/2003 Associate Professor of Pediatrics,


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Biochemistry and Molecular Biology Herman B Wells Center for Pediatric Research James Whitcomb Riley Hospital for Children Indiana University School of Medicine Cancer Research Institute Indianapolis, Indiana

11/1992-4/1999 Children’s Hospital Los Angeles

USC School of Medicine Attending Physician, Division of Hematology-Oncology, Autologous Bone Marrow Transplant Program, Peripheral Blood Stem Cell Program; Post ABMT Biotherapy/Immunotherapy.

1/1993-4/1999 Physician: Pediatric Hematology-Oncology; Neural Tumors Program; Neuro-

oncology Group; Autologous Bone Marrow Transplant Program, Brain Tumors, Neuroblastoma and Retinoblastoma. Assistant Professor, Department of Pediatrics University of Southern California School of Medicine Los Angeles, California

6/1979 – 7/1985 Graduate/Medical MD/PhD Student Research

Department of Microbiology and Immunology University of Miami School of Medicine, Miami, Florida "Isolation and characterization of Wolinella succinogenes L-asparaginase" J. A. Distasio, advisor

9/1975 – 7/1977 Undergraduate Research Experience

Department of Biology, University of Southern Florida Tampa, Florida "Nitrogen fixation in Rhizobium sp." W. S. Silver, advisor

F. TEACHING 1993 – 1999 Division of Hematology-Oncology Weekly Seminars: Clinical Conferences and

Multidisciplinary Conferences Oncology Course Lecture to Fellows: Started format for daily 10-15 minute

lectures to be given to residents prior to start of work rounds. This is found to be a very effective method of teaching in didactic sense. 1997 – 1999 Co-organizer, Signal Transduction Journal Club, Children’s Hospital Los Angeles 1999-2003 B810 Biochemistry Graduate Course, “Molecular Basis for Signal Transduction” Indiana University School of Medicine, Indianapolis, IN 1999- 2003 Member, Graduate Faculty, Indiana University-Purdue University Graduate Program 2009-2014 Director, Subspecialty Pediatric Fellows Research Forum, UCSD Department of

Pediatrics, Rady Childen’s Hospital (twice monthly meeting where clinical fellows present ongoing research).


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2012-present Annual lecture on: Grant Writing 101 (to junior faculty and postdoctoral fellows in

Department of Pediatrics and Rady Children’s Hospital. 2009-present Division of Pediatric Hematology-Oncology, lectures and journal club presentations to

residents and hematology-oncology fellows G. COMMITTEE SERVICE 1993-1997 Education Committee, Division of Hematology-Oncology Children’s Hospital Los Angeles, Los Angeles, California 1993-1997 Research Committee, Division of Hematology-Oncology Children’s Hospital Los Angeles, Los Angeles, California 1995-1999 Heidelberger Fellowship Committee, USC School of Medicine, Los Angeles, CA 1999-2003 Member of Graduate Faculty, Indiana University-Purdue University, Indianapolis, IN 1999-2003 Scientific Review Committee, Indiana University School of Medicine 1999-2003 Biomedical Research Committee, Indiana University School of Medicine 2007 Search Committee, Vice-Chair for Research, Dept of Pediatrics, Emory University 2006-2007 MD-PhD MSTP Admissions Committee, Emory University School of Medicine 2007-2009 Conflict of Interest Committee (COI), Emory University 2009-2012 MD-PhD MSTP Admissions Committee, UCSD School of Medicine 2010-2013 Academic Review Committee (ARC), UCSD School of Medicine 2011-present iHOPE Genomics Committee, UCSD School of Medicine 2009-present Research Council, UCSD Department of Pediatrics 2010-2013 Committee to plan Annual Department of Pediatrics Retreat 2009-2012 Organizing Committee for Annual Department of Pediatrics Research Symposium 2012-present Member of Moores Cancer Center, Cancer Cabinet 2014-present Co-Director Biorepository Moores UCSD Cancer Center 2012-present Co-Director, Moores Cancer Center, Pediatric Oncology Disease Team 2009-present Member, Moores Cancer Center, Executive Committee 2010-present Member, Committee for junior faculty and clinical translational faculty development 2013-present Member, Protocol review maintenance committee (PRMC) (Scientific review IRB

protocols) 2015-present Member, Center for Microbiome Innovation, UCSD School of Medicine H. SOCIETY MEMBERSHIPS American Association for the Advancement of Sciences American Academy of Pediatrics American Association for Cancer Research (AACR) American Society for Clinical Oncology (ASCO) American Society for Microbiology (ASM) Society for Pediatric Research (SPR) American Society for Hematology (ASH) American Society for Pediatric Hematology-Oncology (ASPHO)

I. NIH STUDY SECTIONS/ADVISORY BOARDS/NATIONAL COMMITTEES


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2000 NIH, NCI Special Emphasis Panel – Molecular Targets for Drug Discovery in Cance 2001 SPORE, Special Programs Research Excellence, Brain Tumors (ad hoc)

2002-2005 RAID/NCI Biologic Oversight Committee for RAID Program (4 year term) 2002 PO1 review “Lipid Mediators in Cancer (ad hoc) (9/2002) 2003 NIH/PPG PO1 review “Glioblastoma, PTEN and Imaging” (ad hoc) (9/2002) 2003 NIH/RFA “Bone-Cancer Cell Interactions” (ad hoc) (4/2003) 2004 NIH/PPG PO1 review “Glioblastoma, PTEN and Imaging” (9/2003) 2003 NIH Study Section ZRG F09 (NIH Fellowship Review) (7/2003) 2003-2007 Tumor Microenvironment Study Section (TME) (member) (4 year term)

2007 P30 Study Section, St Jude Childen’s Research Foundation (renewal) 2008 CTEP NCI task force for PAM (PI-3 kinase-AKT-mTOR) 2015 Member, Scientific Advisory Board, Data4Cure (Leroy Hood, T. Ideker and Napo

Ferrara) J. EDITORIAL BOARD ACTIVITIES AND REVIEW 2000-2003 Managing Editor, Frontiers in Bioscience, Section: “Angiogenesis and

Signaling” 2008-present Editorial Board, International Journal of Oncology, Open Journal of Apoptosis

(OJAp) 2013-present Editorial Board, Journal of Pediatric Oncology 2014-present Editorial Board, Frontiers in Pediatric Oncology 2015-present Editorial Board, Molecular Cancer Research (AACR) 2014-present Editorial Board, Macrophage 2015-present Editorial Board, Recent Patents on Endocrine, Metabolic and Immune Drug

Discovery (PMED) 2015-present Reactome reviewer for PTEN signaling axis 1999-present Oncogene (ad hoc)

Journal of Immunology (ad hoc) Blood (ad hoc)

The Journal of Infectious Diseases (ad hoc) Molecular and Cellular Biology (ad hoc) Journal of Biological Chemistry (ad hoc) Journal of Experimental Medicine (ad hoc) Nature (ad hoc) FEBS letters (ad hoc) Neuro-Oncology (ad hoc) Cancer Research (ad hoc) Molecular Cancer Research (AACR) Frontiers in Oncology K. EXTRAMURAL GRANTS AND CONTRACTS

Current/Funded

RO1 FD004385-01A2 “Phase II trial of poly-ICLC in pediatric low grade glioma” Principal Investigator Project Period: 7/01/14-6/31/2018


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ADC: $258,000 R41 CA192656-01 “Maximum MYC control using dual PI-3K/BRD4 inhibitors” Principal Investigator Project Period: 9/01/15-8/31/2016 ADC: $209,000 (score 25) R42 CA192656-01 “Maximum MYC control using dual PI-3K/BRD4 inhibitors” Principal Investigator (Phase II STTR) Project Period: 9/01/16-8/31/2018 ADC: $1,998,000 (score 29)

FDA RO1 FD-R-05113 “Phase I trial of dual PI-3K/BRD4 inhibitor SF1126 in hepatocellular carcinoma” Principal Investigator Project Period: 9/01/16-8/31/19 ADC: $250,000 (Score 23) R41 CA206859 “Maximizing synthetic lethality with dual PI-3K/PARP inhibitor” Principal Investigator Project Period: 7/01/16-6/31/2017 ADC: $298,000 Develop dual PI-3K/PARP inhibitor for cancer therapeutics (score 33) Merck LKRD149174 “Role of PD1 in innate immunity, macrophage M1 to M2 transition” Principal Investigator Project Period: 9/01/15-8/31/17 ADC: $100,000

St. Balderick’s Foundation Grant Co-investigator (with NANT consortium) Funding for Phase I trial of SF1126 (First PI-3K inhibitor in pediatric oncology) Project Period: 7/01/14-6/31/18 ADC: $270,000 Faculty mentor and co-investigator on multiple institutional T32 and K12 training grants at UCSD e.g. “UCSD training program in drug development” T32-121938, PI: Howell.

Pending/Recently submitted or resubmitted

RO1 RO1 “Genome driven Phase II trial of SF1126 in PIK3CA mutated SCCHN” Principal Investigator Project Period: 9/01/16-8/31/2021 ADC: $250,000 x 5 years

RO1 CA204921 “Determine role of Syk in M1-M2 macrophage transition and metastasis” Principal Investigator Project Period: 9/01/16-8/31/2021 ADC: $296,000


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RO1 CA172513-01-A1 “Role of PTEN and PI-3 kinase in medulloblastomagenesis” Principal Investigator Project Period: 9/01/16-8/31/2021 (score 37) ADC: $233,000

RO1 HL108838 “Transcriptomic analysis of sickle cell disease leukocytes; target discovery” Principal Investigator Project Period: 9/01/16-8/31/2021 ADC: $250,000 (score: 27)

Recent & Past

Hyundai Hope Grant Principal Investigator Project Period: 10/01/13-9/01/15 ADC: $125,000 UCSD Department of Pediatrics Genomics Grant “Genomics of Sickle Cell Disease” Principal Investigator Project Period: 5/01/13-4/31/15 ADC: $50,000

R21CA173330-A1 “Nanoparticle enabled L-asparaginase therapy for CLL” Co-investigator (Esener) Project Period: 7/01/13-6/30/15 ADC: $250,000

RO1 CA94233-09 “Vascular targeted PI-3 kinase inhibitor, SF1126 for glioma therapy” Principal Investigator Project Period: 9/01/09-6/31/2013 ADC: $250,000

R21HL091385-01 “Role of Syk and rac2 in regulation of HIF1 and neovascularization”

Principal Investigator Project Period: 9/01/09-8/31/2011

National Institutes of Health ADC: 150,000

R56 CA94233-06 “Vascular targeted PI-3 kinase inhibitor for glioma therapy” Principal Investigator (NDBA) Project Period: 9/01/08-8/31/2009 ADC: $233,000

Alex Lemonade Stand Springboard Grant Principal Investigator Project Period: 9/01/13-8/31/14 ADC: $100,000 Alex Lemonade Stand Foundation Principal Investigator


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Project Period: 7/30/08-6/30/2010 ADC: $100,000

Cure Childhood Cancer Principal Investigator Project Period 7/01/07-6/31/2009 ADC: $213,000 Magic Water Foundation Award Principal Investigator Project Period 3/01/08-2/28/2009 ADC: $100,000

Goldhirsh Foundation Award Principal Investigator Project Period: 7/01/07-6/30/2008 ADC: $100,000

Georgia Cancer Coalition Scholar Award Project Period 9/27/03-8/31/08

ADC: $150,000

Co-Principal Investigator Multiple Myeloma Research Committee “SF1126 Multiple Myeloma into the Clinic” Project Period 1/31/07 to 12/31/2008 ADC: $95,454

Principal Investigator EntreMed Pharmaceuticals “Role of PTEN and PI-3 kinase in 2ME2 therapeutics” Project Period 1/31/07-12/31/2008 ADC: $147,800

Co-Investigator (PI MC Dinauer) NIH RO1 “Phagocyte NADPH Oxidase and Signaling” Project Period: 12/1/01-11/31/06 Annual Direct Cost: $250,000

Co-Investigator (PI J Garlich) NIH R43 SBIR “RGD Chelate Chemistry for Molecular Targeting” Project Period 9/1/01-8/31/03 Annual Direct Cost: $250,000 Co-investigator (PI J Garlich) NIH R43 SBIR “Targeted p53 inhibitors molecular therapeutics” Project Period 4/1/02-3/31/03 ADC: $166,000


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Co-investigator (PI J Garlich) NIH R43 SBIR “Bone targeting degradable drug filled nanoparticles” Project Period 4/1/03-3/31/05 ADC $248,000

Principal Investigator (Leader of Project 2)

“Biology and Therapy of High Risk Neuroblastoma” PO1 CA81403 Project 2: “Vascular Endothelial Cell Integrins v3 and v5 in neuroblastoma and angiogenesis” Program Project Grant/NIH PO1 Project Period: 4/01/00-3/31/04 Annual Direct Cost: $113,922 Principal Investigator NIH NCI RAID “Recombinant Wolinella asparaginase for treatment of ALL” Project Period: 5/1/02-6/31/03 Principal Investigator American Cancer Society "Cbl-adapter Protein Interaction in regulation of RAS" RPG-98-244-01-LBC Project Period: 7/01/98-6/30/02 Annual Direct Cost: $93,342 Annual Indirect Cost: $23,311

Principal Investigator

American Cancer Society Institutional Research Grant (through University of Southern California) IRG-21-34-04 "Generation of Chimeric Receptors Using the Intracytoplasmic Antigen Receptor Homology I Domain" Project Period: 01/01/94-12/31/94 Annual Direct Cost: $14,445 Co-Principal Investigator (Program Project Grant) NIH/UP1, (RFA) CA95-07 “Role of v3 in Tumor Angiogenesis" 4/1/96 – 5/1/99 Annual Direct Cost: $10,544

Co-Principal Investigator “Treatment of Collagen Induced Arthritis with L-Asparaginase” Arthritis Foundation (National) Project Period: 7/01/98-5/1/99 Annual Direct Cost: $80,000 Principal Investigator “An anti-angiogenic approach to treatment of pediatric brain tumors; polyICLC” NIH/NCI RO1 CA75637-01 Project Period: 08/01/97-07/31/00 Annual Direct Cost: $93,736


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Annual Indirect Cost: $46,360

L. INTRAMURAL FUNDING Past 2011-2012 "PolyICLC for pediatric low grade glioma, Phase II trial."

Moores UCSD Cancer Center Pilot Project Award, P30 CA23100-27 Annual Direct Cost: $50,000

12/95-11/98 "GTPases and Raf-1 in mammalian signaling."

Children’s Hospital Los Angeles Career Development Award Annual Direct Cost: $50,000

7/96-6/99 Wright Foundation Award, USC School of Medicine Cbl-adapter protein interaction in Bcr/Abl transformation Principal Investigator Annual Direct Cost: $49,600

1993-1994 "Amino Acid Sequence Requirements for Signal Transduction through FcRI"

Children’s Hospital Los Angeles Intramural Research Grant Annual Direct Cost: $20,500

M. PRIVATE GRANT SUPPORT Current 9/15 – 8-17 Olivia Hudson Foundation (OHF) ADC: $20,000 5/14 – 4/15 Cricket Corporation, Inc. Annual Direct Cost: $75,000 Past 6/93 - 9/95 Pediatric Brain and Solid Tumors

University of Southern California, Gene Therapy Co-Investigator Annual Direct Cost: $70,100

1993-1999 Co-investigator

Cellular & Molecular Biology of Pediatric Cancer Program T.J. Martell Foundation (Program Project) Annual Direct Cost: $54,199

1997-2000 Stop Cancer Research Career Development Award

Stop Cancer Foundation “Anti-angiogenic therapy for treatment of childhood cancer”


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Project Period: 12/01/97 - 11/30/00 Annual Direct Cost: $100,000

N. CLINICAL-TRANSLATIONAL RESEARCH INTERESTS: Neuro-oncology, anti-angiogenic therapies for treatment of cancer and inflammation, Immunoreceptor/ITAM/ITIM signaling, inflammation, graft rejection, GVHD and autoimmune diseases. Small drug-design and biologic therapies for above based on specific signaling pathways. Molecular targets for Drug Discovery. Target discovery and drug development; pharmacokinetic and pharmacodynamic markers and clinical trial development for novel targeted therapeutic agents. Phase I and Phase II trials. Clinical Protocols: Open for enrollment or completed: Poly-ICLC in Management of Recurrent Low Grade Gliomas (Phase II). Donald L. Durden, M.D., Ph.D., Principal Investigator (multicenter trial)(Biology study: TLR3 stimulation of innate and adaptive immune response against the tumor is the mechanism for polyIC antitumor activity). This is form of immunotherapy under study in the Durden laboratory (open to enrollment, Clintrials.gov: NCT01188096). RO1 FDA FD-04385 grant funded 2014, $1.6M, (IRB 101780). 11 patients enrolled, 50% response rate so far. https://www.clinicaltrials.gov/ct2/results?term=polyICLC+and+pediatrics&Search=Search Vascular targeted pan PI-3 kinase bromodomain inhibitor, SF1126, for neuroblastoma therapeutics. First in pediatrics Phase I trial in pediatric solid tumor. NANT Phase I trial. Approved. Durden, inventor and developer of this agent. (opened for enrollment 8/2015). https://www.clinicaltrials.gov/ct2/show/NCT02337309?term=SF1126&rank=2 Poly-ICLC in Management of Newly Diagnosed Malignant Gliomas and Recurrent Pediatric Brain Tumors (Phase II). Donald L. Durden, M.D., Ph.D., Principal Investigator (multicenter trial)(Biology study: TLR3 agonist activates innate and adaptive immunity as mechanism for polyIC antitumor activity) (closed to accrual) (RO1 CA75637) (reported, J. Ped Hem-Onc, 2012). Rapamycin and VP-16 in recurrent solid tumors (Phase I). Terry Vik and D. Durden (Principal Investigators) (closed to enrollment). Use of targeted pan PI-3 kinase inhibitor, SF1126 for the treatment of adult cancer (Phase I). Dan Von Hoff, M.D., Principal Investigator, TGEN-US Oncology and Indiana University School of Medicine (multicenter trial adult solid tumor) (SF1126 Phase I started 4-2007)(closed to enrollment) (reported: Eur.J. Cancer, 2012). Use of targeted pan PI-3 kinase inhibitor, SF1126 for the treatment of refractory multiple myeloma and B cell malignancies (Phase Ib). Saga Lonial, M.D., Principal Investigator, Emory University School of Medicine, Arizona Cancer Center (multicenter trial)(MMRF)(Accrual now 8 patients)(closed to enrollment) (reported Eur. J. Cancer, 2012; CCP, 2013). Whole transcriptome analysis of leukocytes in patients with sickle cell disease undergoing vasoocclusive crisis compared to baseline leukocytes sorted from same patient. Jenny Kim, M.D., Principal Investigator, UCSD Rady Children’s Hospital (multicenter study in pediatrics) (Co-Investigator) RO1 and U54 submitted 2012 (open to enrollment). RNAseq of monocytes reveal M1 vs M2 skewed signature associated with crisis. Clinical Protocols in Development:


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PI-3 kinase inhibitor for brain tumor therapeutics (Phase I trial) (Novartis). First in pediatric Phase I trial of BKM120 pan PI-3 kinase inhibitor. Durden is Chair of Biology and Co-Investigator (Approved by CTEP) (PBTC trial to open 2015). Phase I trial of Pembroluzumab (anti-PD1 antibody) in pediatric solid malignancies. First PD1 antibody trial in pediatric oncology. Merck sponsored trial. Co-investigator on this trial. Phase II trial of Pembroluzumab with polyICLC in recurrent pediatric solid tumors. This trial has evolved from our pediatric polyICLC Phase II trial open for low grade glioma patients and ongoing experiments in the Durden laboratory support this concept. Pediatric Pre-emptive precision oncology protocol (P3-Oncology). High risk patients to enroll on-study at diagnosis for molecular profiling using whole exome and immunome sequencing combined with Oncoscan and systems biologic analysis. At relapse enrolled on targeted therapy trials e.g. SF1126 if meets criteria set by molecular tumor board, and bioinformatics platform: www.Data4Cure.com. Use of recombinant Wolinella succinogenes asparaginase for the treatment of leukemia (Phase I/II trial) Frank Keller, M.D., Principal Investigator. (Official COG biology study) NCI RAID supported (going to IRB) (FDA on clinical hold pending new toxicology studies). First recombinant asparaginase. Rapamycin and VP-16 in recurrent neuroblastoma (Phase Ib). Terry Vik and Donald Durden (Principal Investigators) (closed to accrual). Use of a c-raf multikinase inhibitor, Sorafenib with vinblastine for the treatment of recurrent Neuroblastoma (Phase Ib). Lisa Hartman, M.D., Principal Investigator, UCSD Rady Children’s Hospital (multicenter trial in pediatrics). Use of a c-raf multikinase inhibitor, Sorafenib with proteasome inhibitor for the treatment of recurrent Medulloblastoma (Phase Ib). Lisa Hartman, M.D. and John Crawford, MD, Principal Investigators, UCSD Rady Children’s Hospital (multicenter trial in pediatrics). O. PATENTS: Cloning and expression of recombinant Wolinella succinogenes asparaginase for the treatment of hematologic and autoimmune diseases. (provisional patent filed 6/4/97; PCT International Patent & Utility filed 6/9/98). D. L. Durden, Inventor. In preparation for the Phase I/II Clinical Trials. (RAID NCI funded) (U.S. Patent No. 6,255,388) (licensed to Rare Disease Therapeutics)(In production for Phase I trial). PTEN and PI-3 kinase inhibitors and other kinase/phosphatase inhibitors and agonists for treatment of diseases which benefit from control of PI-3 kinase and PTEN signaling pathways. (provisional filed 5-99) D. L. Durden, Inventor (US Patent No. 6,668,002) (licensed to SignalRx Pharmaceuticals). PI-3 kinase inhibitor prodrugs for therapeutic applications. Garlich J. and Durden D.L. Inventors. (September 27, 2005) (U.S. Patent No. 6,958,300) (FDA/IND issued on SF1126 on 1-28-2007). PTEN inhibitors for therapeutic applications. D.L. Durden and Garlich, J. Inventors (United States PI-3 kinase inhibitor prodrugs. Garlich, J. and Durden, D.L. Inventors (US Patent No. 7,396,828) (Issued July 8, 2008).


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PI-3 kinase inhibitor prodrugs. Garlich, J. and Durden, D.L. Inventors (US Patent No. 7,662,997) (Issued Feb, 16, 2010). PI-3 kinase inhibitor prodrugs. Garlich, J. and Durden, D.L. Inventors (US Patent No. 7,666,333) Compositions and methods for identifying agents which modulate PTEN function and PI-3 kinase pathways. D.L. Durden, Inventor (United States Patent Number 7,745,485) (Issued on June 29, 2010). Thienopyranone scaffold as dual PI-3 kinase bromodomain inhibitory chemotypes (Provisional patent filed 4/2014) D. L. Durden, Garlich and Morales inventors. Thienopyranones as kinase inhibitors (continuation) US 8,557,807, filed 9/30/2013. XTDW004YS-A. D. L. Durden, Garlich and Morales inventors. Thienopyranones as Kinase and Epigenetic inhibitors filed 5/5/2014, pending review. D. L. Durden, Garlich and Morales inventors. PTDW035, 61-988346, provisional. Novel Heterocyclic compound classes for signal modulation filed 5/4/2015, pending. D. L. Durden, Garlich and Morales inventors. XTDW034. Thienopyranones as kinase and epigenetic inhibitors filed 5/4/2015, pending. D. L. Durden, Garlich and Morales inventors. P. INDUSTRIAL PARTNERSHIPS: Semafore Pharmaceuticals, Indianapolis, IN. Scientific co-founder and former chairman of the Scientific Advisory Board, working toward the development of small molecule inhibitors of key kinase and phosphatase signaling pathways for application to pediatric and adult diseases, Signal transduction therapeutics. Translation “bench to bedside” First PI-3 kinase inhibitor to enter human clinical trials. Rare Disease Therapeutics, Nashville, TN. Consultant, working toward the development of novel asparaginase to treat acute lymphoblastic leukemia in children. In addition, the development of other orphan drugs targeted to pediatric diseases based on signal transduction therapeutics. Biologica, LLC., Atlanta,GA. Small molecule drug discovery company founded by D. Durden, 2006. Targets include PTEN, PI-3 kinase, Syk, CSK kinases and a number of other phosphatases. This group is involved in: in silico molecular modeling, high through-put screens and high-content screens, chemistry and lead optimization. Other targets are in development. Expertise includes drug development strategies (GMP, GLP, toxicology), target validation in vitro and in vivo and PK and PD developments needed for Phase I and Phase II clinical trial design. SignalRx Pharmaceuticals, Inc., San Diego, CA (Founder) Small molecule drug discovery and development company focused on PI3K and dual inhibitory chemotypes e.g. PI3K/BRD4, PI3K/PARP, PI3K/MEK, etc. inhibitors for cancer therapeutics and PTEN inhibitor drug discovery. The group is focused on rationale drug design of dual inhibitor chemotypes. They developed the first know PI-3K/BET bromodomain inhibitory chemotype. The IP from the Semafore portfolio incorporated into this entity in 2012. Merck, Inc. A partnership with Merck to develop next generation checkpoint inhibitors and to perform pediatric PD1 monoclonal antibody immunotherapeutic trials in solid malignant tumors (PI on Phase II).


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Q. BIBLIOGRAPHY

Peer Reviewed (total of 68 manuscripts) Charyulu, V., Sigel, M.M., Durden, D.L. and Lopez, D.M. Mouse mammary tumor virus (MMTV) antigen(s) are present of B-lymphocytes of Balb/c mice. Int. J. Cancer 24:813-818, 1979. Durden, D.L. and Distasio, J.A. Comparison of the immunosuppressive effects of L-asparaginases from Escherichia coli and Vibrio succinogenes. Cancer Res, 40:1125-1129, 1980. Durden, D.L. and Distasio, J.A. Characterization of the effects of Escherichia coli asparaginase and a glutaminase-free asparaginase from Vibrio succinogenes on specific cell-mediated cytotoxicity. Int. J. Cancer, 27:59-65, 1981. Distasio, J.A., Durden, D.L., Paul, R.D. and Nadji, M. Alteration in spleen lymphoid populations associated with specific amino acid depletion during L-asparaginase treatment. Cancer Res, 42:252-258, 1982. Distasio, J.A., Salazar, A.M., Nadji, M. and Durden, D.L. Glutaminase-free asparaginase from Vibrio succinogenes: an antilymphoma enzyme lacking hepatotoxicity. Int. J. Cancer, 30:343-347, 1982. Durden, D.L., Salazar, A.M. and Distasio, J.A. Kinetic analysis of hepatotoxicity associated with antineoplastic asparaginases. Cancer Res, 43:1602-1605, 1983. Kazlauskas, A., Durden, D.L. and Cooper, J.A. Functions of the major tyrosine phosphorylation site of the PDGF receptor ß subunit. Cell Regul, 2:413-425, 1991. Durden, D.L., Rosen, H. and Cooper, J.A. Serine/threonine phosphorylation of the -subunit after activation of the high-affinity Fc receptor for immunoglobulin G. Biochem, J. 299:569-577, 1994. Durden, D.L., Rosen, H., Michel, B.R. and Cooper, J.A. Protein tyrosine phosphatase inhibitors block myeloid signal transduction through the FcRI receptor. Exp Cell Res, 211:150-162, 1994. Durden, D.L., and Liu, Y.B. Protein-tyrosine kinase p72syk in FcRI receptor signaling. Blood, 84:2102-2108 (Rapid Communication), 1994. Durden, D.L., Kim, H.M., Calore, B., and Liu, Y.B. The FcRI receptor signals through the activation of hck and MAP kinase. J. Immun, 154:4039-4047, 1995. Arditi, M., Zhou, J., Martine, T., Durden, D., Stins, M., and Kwang, S-K. Lipopolysaccharide stimulates the tyrosine phosphorylation of mitogen-activated protein kinases, p44, p42, and p38 in vascular endothelial cells in a soluble CD14-dependent manner: Role of protein tyrosine phosphorylation in lipopolysaccharide-induced stimulation of endothelial cells. J. Immun, 155:3994-4003, 1995. Park, R.K, Liu, Y.B., and Durden, D.L., A role for Shc, Grb2 and Raf-1 in FcRI signal relay. J. Biol. Chem, 271:13342-13348, 1996. Taylor, N., Jahn, T., Smith, S., Lamkin, T., Uribe, L., Liu, Y-B, Durden, D.L. Weinberg, K. Differential activation of the tyrosine kinases ZAP-70 and Syk after FcRI stimulation. Blood, 89:388-396, 1997 (Rapid Communication).


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Erdreich-Epstein, A., Liu, M. Liu, Y.B. and Durden, D. L. Protein tyrosine phosphatase inhibitors in FcRI-induced myeloid oxidant signal transduction. Exp Cell Res, 237: 288-295, 1997 . Park, R.K., Liu, Y.B., Kyono, W., and Durden, D.L. CBL-GRB2 Interaction in myeloid immunoreceptor tyrosine activation motif signaling. J. Immunol, 160:5018-5027, 1998.

Chu, J., Liu, Y, Koretzky, G.A. and Durden, D.L. SLP-76-CBL-Grb2-Shc interactions in FcRI signaling.

Blood, 92:1697-1706, 1998 Kyono, W. T., De Jong, R., Park, R. K., Liu, Y.B., Heisterkamp, N., Groffen, J. and Durden, D.L. Differential interaction of Crkl with Cbl or C3G, Hef-1 and -subunit ITAM in myeloid FcRI signaling. J Immunol, 161:5555-5563, 1998.

Izadi, K., Erdreich-Epstein, A., Liu, Y., and Durden, D.L. Characterization of Cbl-Nck and Nck-Pak1 interactions in myeloid FcRII signaling. Exp Cell Res, 245:330-342, 1998. Erdreich-Epstein, A., Liu, M. Kant, A., Izadi, K., Nolta, J. and Durden, D.L. CBL functions downstream of Src kinases in FcRI signaling in primary human macrophages. J Leuk Biol, 65:523-534, 1999. Park, Rae-Kil, Izadi, K., Deo, Y.M., Liu, YB and Durden, D.L. Role of Src in the modulation of multiple adaptor proteins in FcRI oxidant signaling. Blood, 94:2112-2120, 1999. Park, R.K., Erdreich-Epstein, A., Liu, M., Izadi, K. D., and Durden, D.L. High Affinity IgG Receptor Activation of Src Family Kinases Is Required for Modulation of the Shc-Grb2-Sos Complex and the Downstream Activation of the Nicotinamide Adenine Dinucleotide Phosphate (Reduced) Oxidase. J Immunol, 163:6023-6034, 1999. Erdreich-Epstein, A., Shimada, H., Groshen, S., Liu, M., Metelitsa, L., Kim, K.S., Stins, M., Seeger,R.C. and Durden, D.L. Integrins v3 and v5 are expressed by endothelium of high-risk neuroblastoma and their inhibition is associated with increased endogenous ceramide. Cancer Research, 60:712-721, 2000. Wen, S., Stolarov, J., Myers, M.P., Su, J.D., Wigler, M.H., Tonks, N.K. and Durden, D.L. PTEN controls tumor-induced angiogenesis. PNAS. 98: 4622-4627, 2001. Isogai, C, Laug, W.E., Shimada, H., Declerck, P., Stins, M. Erdreich-Epstein, A., Durden, D.L., De Clerck, Y.A.. PAI-1 Promotes Angiogenesis by Stimulating Endothelial Cell Migration Toward Fibronectin. Cancer Research. 61: 5587-5594, 2001. Reif, A. and Durden, D.L. Treatment of collagen induced arthritis in DBA/1 mice with L-asparaginase. Clin Exp Rheumatol. 19: 639-646, 2001. Mayo, L.D., Dixon, J.E., Durden, D.L., Tonks, N.K. and Donner, D.B. PTEN protects p53 from MDM2 and sensitizes cancer cells to chemotherapy J. Biol.Chem. 277: 5484, 2002. Kim, J.S. Peng X. De, P.K. Geahlen, R.H. and Durden, D. L. PTEN controls immunoreceptor(ITAM) signaling and the activation of Rac. Blood, 99: 694, 2002. Kant, A., De, P.K., Kim, J.S., Rawlings, D.A., Yi, T., and Durden, D.L. Tyrosine phosphatase SHP-1 regulates Fc receptor mediated phagocytosis and activation of RAC in myeloid cells. Blood, 99: 1879, 2002.


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Erdreich-Epstein, A., Millard, M., Tran, L.B., Wang, H., Cabot, M.C., Durden, D.L., Frgala, T., Reynolds, D.P., Stins, M.F., Bowman, N.N. Molecular ordering of ceramide signaling in fenretinide induced endothelial apoptosis. J.Biol.Chem. 277: 49531, 2002. Su, J.D., Mayo, L.D., Donner, D.B. and Durden, D.L. PTEN and PI-3 kinase inhibitors upregulate p53 and block tumor induced angiogenesis; Evidence for an effect on the tumor and endothelial compartment Cancer Res. 63: 3585-3592, 2003. De P. Peng, X. and Durden, D.L. Rac2 specificity in macrophage integrin signaling: Potential role for Syk kinase J. Biol.Chem. 278: 41661-41669, 2003. Moon, K.D., Post, C.B., Durden, D.L., Zhou, Q., De, P., Harrison, M.L. and Geahlen, R.L. Molecular basis for a direct interaction between Syk protein tyrosine kinase and PI-3 kinase. J. Biol. Chem. 280: 1543-1551, 2004. Rong, Y., Post, D.E., Pieper, R.O., Durden, D.L., Van Meir, E.G. and Brat, D.J. PTEN and hypoxia regulate tissue factor expression and plasma coagulation by glioblastoma. Cancer Research 65: 1406-1423, 2005. Dey, N., Howell, B.J. De, P. and Durden, D.L. CSK controls NGF induced neuronal differentiation and activates AKT. Exp. Cell Res. 307: 1-14, 2005. Rong, Y., Hu, F., Huang, R., Mackman, N., Horowitz, J., Jensen, R., Durden, D.L., VanMeir, E., Brat, D. DGR-1 Regulates Hypoxia-induced Expression of tissue Factor in Glioblastoma through HIF-1 Independent Mechanisms., Cancer Res. 66: 7067-7074, 2006. Rong Y, Durden DL, Van Meir EG, Brat DJ., 'Pseudopalisading' necrosis in glioblastoma: a familiar morphologic feature that links vascular pathology, hypoxia, and angiogenesis. J Neuropathol Exp Neurol. 65(6):529-39, 2006 Reinert, R.B., Oberle, L.M., Wek, S.A., Bunpo, P., Wang, S.P., Mileva, I., Goodwin, L.O., Aldrich, C.J., Durden, D.L., McNurlan, M.A., Wek, R.A. and Anthony, T.G. Role of glutamine depletion in directing tissue-specific nutrient stress responses to Asparaginase. J. Biol.Chem. 281: 31222-31233, 2006. Denning, G., Prince, C., Jean-Joseph, B., Durden, D.L. Vogt, P.K., Nuclear localization as a mechanism for inhibiting the tumor suppressor properties of PTEN., Oncogene, 26: 3930-3940, 2007 (equal senior author contribution). Dey, N., De, P.K., Wang, M., Zhang, H.Y., Dobrata, E.A., Robertson, K. and Durden, D.L. CSK controls retinoic acid receptor signaling: RAR-cSRC signaling axis is required for neuritogenic differentiation. MCB, 27: 4179-4197, 2007. Yu, L., Yan, J., De, P., Chang, H-C., Yamauchi, A., Christopherson, K.W., Peng, X., Kim, C., Kapur, R., Stone, J.C., Kaplan, M.H., Dinauer, M.C., Durden, D.L. and Quilliam, L.A. Rap1a null mice have altered myeloid cell functions suggesting distinct roles for the closely related Rap1A and 1B proteins. J. Immunology, 179: 8322, 2007. Garlich, J.R., De. P., Dey. N., Su, J.D., Mills, G.B., Peng, Q. and Durden, D.L. A vascular targeted PI-3 kinase inhibitor prodrug with antitumor and antiangiogenic activity Cancer Res. 68: 206, 2008.


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Dey, N., Crosswell, H., De. P., Su, J D., Parsons, R., and Durden, D.L. PTENs protein phosphatase activity regulates Src family kinases and controls glioma migration. Cancer Res. 68: 1862, 2008. De, P., Su, J.D., Yoder, M.C. and Durden, D.L. A blood specific GTPase, Rac2 encodes the postnatal neovascular response. Exp.Cell Research, 2009:315, 248. Rong, Y., Belozerov, V.E., Tucker-Burden, C., Chen, G., Durden, DL, Olson, JJ., Van Meir, EG and Brat, D. EGFR and PTEN modulate tissue factor expression in glioblastoma through JunD/AP-1 transcriptional activity. Cancer Res. 69, 2540, 2009. Crosswell, H.E., Dasgupta, A., Alvarado, C.S., Watt, T., Christensen, J.G., De, P. Durden, D.L. Findley, H.W. PHA665752, a small molecule inhibitor of c-Met inhibits hepatocyte growth factor stimulated migration and proliferation of c-Met positive neuroblastoma cells. BMC Cancer, 9: 411, 2009, PMCID PMC2790467. Gu, L., Zhu, N. Zhang, HY, Durden, DL, Feng, Y. and Zhou, MZ. A novel role for mdm2 in the regulation of XIAP protein translation and apoptosis. Cancer Cell, 15, 363, 2009. Ozbay, T., Durden, D.L., Liu, T., O’Regan, R.M. and Nahta, R. In vitro evaluation of pan PI-3 kinase inhibitor SF1126 in trasuuzumab-sensitive and trastuuzumab-resistant HER2 overexpressing breast cancer cells. Cancer Chemother. Pharmacol., 65: 697, 2010. Castellino, R.C., Barwick, B.G., Schniederjan, M., Wagar, N., Liu, J., Brat, D.J and Durden, D.L. Heterozygosity for Pten promotes tumorigenesis in a mouse model of medulloblastoma PLoS One. 26: 10849, 2010. Peirce, S.B., Findley, H.W.,Cooper, T., Dasgupta, A. and Durden, D.L. The PI-3 kinase-Akt-MDM2-survivin signaling axis in high risk neuroblastoma: a target for PI-3 kinase inhibitor intervention. Cancer Chemother. Pharmacol., 68: 325, 2011. Mahadevan, D., Chiorean, E.G., Harris, W.B., Von Hoff, D.D., Stejskal-Barnett, A. Qi, W., Anthony, S.P., Younger, A.E., Rensvold, D.M., Cordova, F., Shelton, C.F., Becker, M.D., Garlich, J.R., Durden, D.L. and Ramanathan, R.K. A phase I clinical trial of a vascular targeted pan PI3K inhibitor, SF1126 in solid tumors and B cell malignancy. Eur .J. Cancer, 48: 3319, 2012. Emmenegger, B.A., Hwang, E., Moore, C., Markant, S.L. Brun, S.N., Dutton, J.W., Read, T.A, Fogarty, M.P., Singh, A., Durden, D.L., Yang, C., McKeenan, W.L. and Wechsler-Reya, R.J. Distinct roles for fibroblast growth factor signaling in cerebellar development and medulloblastoma, Oncogene, 32: 4181, 2012. Qi, W., Stejskal, A., Morales, C., Cooke, L.S., Garlich, J.R., Mahadevan , D. and Durden D.L. SF1126, a pan-PI3K inhibitor has potent pre-clinical activity in aggressive B-cell non-hodgkin lymphomas by inducing cell cycle arrest and apoptosis. J. Cancer Sci Ther, 2012. De, P., Dey, N., Terakedis, B., Bersagel, L., Trudel, S., Makale, M., Garlich, J.R., Durden, D.L. An integrin-targeted, pan phosphoinositide-3 kinase inhibitor, SF1126, has activity against multiple myeloma in vivo. Cancer Chemother. Pharmacol. 74: 867, 2013. Morales, G., Garlich, J.R. Su, J.D., Peng, X-D., Newblom, K., Weber, K. and Durden, D.L. Synthesis and cancer stem cell-based activity of substituted 5-morpholino-7H-thieno [3,2-b]pyran-7-ones designed as next generation PI3 kinase inhibitors. J. Med. Chem., 56: 1922, 2013.


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Hartman, L., Crawford, J.R., Makale, M. Milburn, M., Joshi, S., Salazar, A.M., Hasenauer, B., MacDonald, T.J. and Durden D.L. A Phase II trial of PolyICLC in the management of newly diagnosed and recurrent pediatric brain tumors. J. Ped. Hem. Oncology, 36: 451, 2013. Muh, C.R., Joshi, S., Singh, A.R., Kesari, S., Durden, D.L. and Makale, M.T. PTEN status mediates 2ME2 anti-tumor efficacy in preclinical glioblastoma models; role of HIF1 suppression. J. Neurooncology, 116: 89, 2013. Joshi, S., Singh, A.R., Zulcic, M., Bao, L., Messer, K., Ideker, T., Dutkowski, J. and Durden, D.L. Rac2controls tumor growth, metastasis and M1 to M2 macrophage differentiation in vivo. PLoS One. 25: e95893, 2014. Singh, A.R., Peirce, S.B., Joshi, S., and Durden, D.L. PTEN and PI-3 kinase inhibitors control LPS signaling and lymphoproliferative response in the CD19+B cell compartment. Experimental Cell Research. 327: 78, 2014. Joshi, S., Singh, A.R., Zulcic, M. and Durden, D.L. PTEN/PI-3 kinase signaling axis regulated hypoxia induced HIF1 and HIF2 stability in macrophages and controls tumor growth, angiogenesis and metastasis. Mol. Canc. Res. 10: 1520, 2014. Joshi, S., Singh, A.R., Zulcic, M. and Durden, D.L. PKC desensitizes Fc receptor signaling and leads to inhibition of phagocytosis through an activation of protein tyrosine phosphatase, SHP-1 action. BMC Immunology. 15: 18, 2014. Joshi, S., Singh, A.R. and Durden, D.L. MDM2 regulates hypoxic HIF1 stability in an E3 ligase. Proteasome and PTEN-PI-3 kinase-AKT dependent manner. J. Biol. Chem., 289: 22785, 2014. Joshi, S., Singh, A.R. and Durden, D.L. Pan-PI-3 kinase inhibitor SF1126 shows antitumor and antiangiogenic activity in renal cell carcinoma. Cancer Chemother Pharmacol., 75: 595,2015. Bhat, Vikas, Merissa Olmer, Shweta Joshi, Donald L Durden, Thomas Cramer, Richard Barnes, Scott Ball, Tudor H Hughes, Mauricio Silva, James V Luck, Laurent O Mosnier, Martin Lotz & Annette von Drygalski. Vascular Remodeling underlies re-bleeding in hemophilic arthropathy. Am. J. Hematol, In Press, 2015. Yuki Ishii; May Keu Nhiayi; Jonathan Cheng; Michele Massimino; Donald L Durden; Paolo Vigneri; Jean Y J Wang, Knockout Serum Replacement Promotes Cell Survival by Preventing BIM from Inducing Mitochondrial Cytochrome C Release, PLoS One, In Press, 2015. Singh, A.R., Joshi, S., Garlich, J.R., Morales, G,A., Cho, Y-J., Bao, L., Messer, K.X., Levy, M.L., Crawford, J.R. and Durden, D.L. Role of PTEN and PI-3 kinase in CD15+ cancer stem cell dependent medulloblastoma therapeutics. PLoS One. 2016, In Press. Books and Reviews Dey, N., Durden, D.L. and Van Meir, E.G. Cytokines expression and signaling in brain tumors. In: Cytokines in the CNS (2nd edition) Eds Ransohoff, R.M. and Benveniste, E.N., CRC Press, 2005. Castellino, RC and D.L. Durden PTEN/PI-3 kinase signaling node: An intercept point for the control of Angiogenesis Nature Clin Practice Neurology 3: 682, 2007 (Invited review).


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Castellino, RC, Muh, C.R. and Durden D.L. PTEN and the coordination of angiogenesis, Current Pharmaceutical Design, 15: 380 (Invited review) 2009. Joshi, S. Singh, A., Hartman, L., Ahn, H., Zulcic, M. and Durden, D.L. New therapeutic approaches for Neuroblastoma, In: Neuroblastoma, Ed H. Shimada, Intech, 2013 (In press). “BET bromodomain targets in cancer” Durden, D.L., and McClure, J., Editors, Springer books, (Invited to serve as editors for this volume) (In preparation for 2016 release). Manuscripts Submitted: Joshi, S., Singh, A.R., Zulcic, M., Gosselin, D., Glass, C.K., and Durden, D.L. Unexpected role of BRD4 in the regulation of macrophage M1-M2 transition and metastasis; epigenetic reprogramming the M2 transition. PNAS (submitted). Chattopadhyay, M., Fanale, S., and Durden, D.L. PTEN controls DNA repair and regulates sensitivity to PARP inhibitors and ionizing radiation: Molecular mechanism explored. Cancer Research. (submitted). Shu, H-K, Gao, H., Chang, C-M and Durden, D.L. The pan PI-3 kinase inhibitors SF1126 and SF2523 radiosensitize human glioma cells and impairs the repair of radiation-induced DNA double stranded breaks. Cancer Chemother. Pharmacol. (submitted). Singh, A.R., Joshi, S., Garlich, J.R., Morales, G,A. and Durden, D.L. Dual PI-3K/PARP inhibitors for maximum synthetic lethality to chemoradiotherapy. Cancer Chemother. Pharmacol. (submitted). Singh, A.R., Burgoyne, A., Joshi, S., Garlich, J.R., Morales, G,A. and Durden, D.L. Dual PI-3 kinase/BRD4 inhibitor for hepatocellular carcinoma therapeutics. Oncotarget. (submitted). Joshi, S., Singh, A.R., Zulcic, M., Gosselin, D., Glass, C.K., and Durden, D.L. Syk kinase regulates macrophage M1-M2 transition and metastasis; epigenetic reprogramming the M2 transition. Mol. Cell (submitted). Epstein, A. Joshi, S., Singh, A.R., Zulcic, M., Seeger, R.C., Shimada, H. and Durden, D.L. Association of high microvessel v3 and low PTEN with poor outcome of Stage 3 neuroblastoma: rationale for the application of an RGD targeted pan PI-3 kinase inhibitor therapeutic agent. Oncotarget (submitted). Manuscripts in Preparation Joshi, S., Singh, A.R., Zulcic, M., Gosselin, D., Glass, C.K., and Durden, D.L. Role of Rac2 in the control of liver and lung fibrosis; Control of M1-M2 transition. PLoS One (In preparation). Joshi, S., Singh, A.K., Wang, C., Ahmad, A., Fanale, S. and Durden, D.L. Antileukemic and immunological characterization of a glutaminase free recombinant Wolinella asparaginase; Structural basis for glutaminase activity. (In preparation). Abstracts Durden, D.L. and Distasio, J.A. 1979. The immunosuppressive effects of asparaginases: A comparative study. Clinical Res. 27:3, 604A.


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Charyulu, V., Durden, D.L. and Lopez, D.M. 1979. Presence of mammary tumor virus (MMTV) antigen(s) in the surface of B-lymphocytes of Balb/c mice: Specificity of the immunofluorescence reaction. In: American Society for Microbiology, Abstracts of the 79th Annual Meeting, p. 39H. Durden, D.L. and Distasio, J.A. 1979. The immunosuppressive effects of asparaginases: A comparative study. In: American Society for Microbiology, Abstracts of the 79th Annual Meeting, p. 109.

Durden, D.L. and Distasio, J.A. 1980. Characterization of antibody-dependent cell-mediated cytotoxic (ADCC) response of specifically immunized murine spleen cells against sheep red blood cells. In: Federation Proceedings, Abstracts of the 64th Annual Meeting, FASEB Press, Bethesda, p. 925. Durden, D.L. and Distasio, J.A. 1980. A comparison of the humoral and cell-mediated immune responses in mice treated with Escherichia coli and Vibrio succinogenes asparaginases. In: American Society for Microbiology, Abstracts of the 80th Annual Meeting, p. 59. Durden, D.L., Pollack, A. and Distasio, J.A. 1980. Characterization of asparaginase-induced immunosuppression of specific antibody-dependent cell-mediated cytotoxicity. In: Abstracts of the 4th International Congress of Immunology, Abstract #17.7.10. Durden, D.L., Salazar, A.M., Nadji, M. and Distasio, J.A. 1982. Vibrio succinogenes asparaginase: An anti-lymphoma agent lacking hepatotoxicity. Proc. Am. Assoc. Cancer Res., 23:204. Durden, D.L., Rosen, H., Michel, B.R. and Cooper, J.A. 1992. Protein tyrosine phosphorylation and the neutrophil respiratory burst. Keystone Symposia on Molecular & Cellular Biology. Abstract No. H219, Keystone CO. Arditi, M., Zhou, J., Durden, D.L., Torres, M., Kim, K.S. 1994. Lipopolysaccharide-induced IL-6 production in vascular endothelial cells: Role of tyrosine phosphorylation in transmembrane signaling. FASEB Journal, 8:A1019. Abstract #5901. 1995. Durden, D.L., Kim, H.M., and Liu, Y.B. 1994. A role for syk and shc in FcRI signal transduction. Tenth Annual Meeting on Oncogenes, Hood College, Frederick, Maryland, 1994. Jahn, T., Taylor, N., Smith, S., Liu, Y., Uribe, L., Durden, D., and Weinberg, K. Differential activation of the tyrosine kinases Zap-70 and syk following FcRI stimulation. Blood 86(10):29a Suppl 1, 1995. Abstract #196. Gonzalez, F.I., Liu, Y.B., Liu, M., and Durden, D.L. Differential effects of growth factors on FcRI signaling through hck, MAP kinase and vav proto-oncogenes. Blood 86(10):694a Sup 1,1995. Abs.#2764. Arditi, M., Zhou, J., Durden, D.L., Liu, Y.B., Torres, M., Kim, K.S. LPS induces activation of MAP kinases and tyrosine phosphorylation of raf-1 and a 66-Kda raf-1-associated protein. Keystone Symposia on Molecular and Cellular Biology, 1996. Erdreich-Epstein, A., and Durden, D.L. Protein tyrosine phosphatases regulate the cbl-grb2 interaction in FcRI signaling. Twelfth Annual Meeting on Oncogenes, Hood College, Fredrick, Maryland, 1996. Park, R.K., Kyono, W.T., Liu, Y., Durden, D.L. Grb2-SH3-SH2-cbl interaction in immunoreceptor tyrosine activation motif (ITAM) signaling. Twelfth Annual Meeting on Oncogenes, Hood College, Fredrick, Maryland, 1996.


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Durden, D.L., Park, R.K., Kyono, W., and Liu, Y.B. Grb2-cbl and Crkl-cbl interaction in the regulation of FcRI signaling and the activation of ras. Tyrosine Phosphorylation and Cell Signaling. The Salk Institute, La Jolla, California, 1996. Kyono, W. T., Park, R. K. Liu, Y. B. and Durden, D.L. CRKL-CBL interactions in myeloid FcRI signal transduction. Meeting American Society for Hematology (ASH), December, 1996. Park, R. K. Kyono, W.T. Liu, Y.B., Chung, H.T. and Durden, D.L. Characterization of SHC-GRB2-CBL interaction in immunoreceptor tyrosine activation motif (ITAM) induced activation of RAS. Temporal and Spacial determinants of specificity in signal transduction, Keystone Symposia on Molecular and Cellular Biology, March,1997. Erdreich-Epstein, A., Liu, Ming, Liu, Y.B., Durden, D.L. Protein tyrosine phosphatase inhibitors in FcRI myeloid signal transduction. Temporal and Spacial determinants of specificity in signal transduction, Keystone Symposia on Molecular and Cellular Biology, March, 1997. Izadi, K. D., Erdreich-Epstein, A. and Durden, D.L. The NCK-CBL interaction in FcRII signaling in myeloid cells. Thirteenth Annual Meeting on Oncogenes, Hood College, Frederick, Maryland, 1997. Erdreich-Epstein, A., Liu, Ming, Liu, Y.B., Koretzky, G. A., Nolta, J. A. and Durden, D.L. CBL is phosphorylated and associated with CRKL, SLP-76, NCK and GRB2 following FcRI stimulation in bone marrow derived primary human macrophages. Thirteenth Annual Meeting on Oncogenes, Hood College, Frederick, Maryland, 1997. Chu, J., Liu, Y.B., Koretzky,G. A., Durden, D.L. SLP-76-CBL-GRB2-SHC interactions in FcRI signaling. Thirteenth Annual Meeting on Oncogenes, Hood College, Frederick, Maryland, 1997. Erdreich-Epstein, A., Song, P., Gonzalez-Gomez, I., and Durden, D.L. Integrin v3 is expressed on angiogenic endothelial cells in malignant pediatric brain tumors. In: The American Society of Pediatric Hematology-Oncology 10th Annual Meeting, San Francisco, California, September 18-20, 1997, pp. 24, Young Investigator Oncology Basic Research Paper Presentation. Erdreich-Epstein, A., Shimada, H., Seeger, R.C., and Durden, D.L. Integrin v3 is expressed on angiogenic endothelial cells in malignant pediatric neural tumors. Eighty-ninth Annual Meeting. American Association for Cancer Research, Abstract# 1028, Poster Presentation, 1998. Metelitsa, L.S., Keshelava, N., Reynolds, C.P., Durden, D.L. and Seeger, R.C. Ceramide induced cytotoxicity in human neuroblastoma cells correlates with their sensitivity to cisplatin, carboplatin, and doxorubicin but not to melphalan and etoposide. Eighty-ninth Annual Meeting. American Association for Cancer Research, Abstract# 473, 1998. Izadi, K., Erdreich-Epstein, A., and Durden, D.L. Characterization of Cbl-Nck and Nck-Pak1 interactions in myeloid signaling. Fourteenth Annual Meeting on Oncogenes, The Salk Institute, La Jolla, California. Abstract #118, Poster Presentation, 1998. Kant, A., Liu, Y.B., Chu, J., and Durden, D.L. The role of Syk kinase, the Cbl-Crkl-C3G and Cbl-SLP-76 interactions in Fc receptor mediated phagocytosis and myeloid signaling. Fourteenth Annual Meeting on Oncogenes, The Salk Institute, La Jolla, CA 1998. Abstract #176. Isogai, C., Erdreich-Epstein, A., Shimada, H., Seeger, R.C., Groshen, S., Durden, D.L., Laug, W.E., DeClerck, Y.A. Overexpression of plasminogen activator (PA) inhibior-1 (PAI-1) in high-risk


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neuroblastoma: a positive role for PAI-1 in angiogenesis. Ninety-first Annual Meeting of the American Association for Cancer Research, The Moscone Center, San Francisco, CA. 2000. Su, J.D., Stolarov, J., Myers, M.P., Wen, S., Wigler, M.H., Tonks, N.K., and Durden, D.L. PTEN controls the growth and angiogenic response of malignant gliomas. Tyrosine Phosphorylation and Cell Signaling, The Salk Institute, La Jolla, CA. 2000. Abstract #60. De, P., and Durden, D.L. PKC desensitizes Fc receptor signaling and leads to inhibition of phagocytosis. Tyrosine Phosphorylation and Cell Signaling, The Salk Institute, La Jolla, CA. 2000. Abstract #82.

Dey, N., Robertson, K.A., and Durden, D.L. CSK is a negative regulator of neural differentiation. Tyrosine Phosphorylation and Cell Signaling, The Salk Institute, La Jolla, CA. 2000. Abstract #84. Kim, J.S., Kant, A., Rawlings, D.A., Yi, T., and Durden, D.L. Tyrosine phosphatase SHP-1 and SHIP regulate Fc receptor mediated phagocytosis in myeloid cells. Tyrosine Phosphorylation and Cell Signaling, The Salk Institute, La Jolla, CA. 2000. Abstract #101. Bowman, N., Durden, D.L. and Erdeich-Epstein, A. Additive effect of fenretinide and the RGD-blocking peptide RGDfV on endothelial cell ceramide and apoptosis. Ninety-second Meeting of the American Association for Cancer Research, New Orleans, LA, 2001 Su J.D., Stolarov, J., Wen, S., Myers, M., Wigler, M.H. Tonks, N.K. and Durden, D.L. PTEN controls tumor-induced angiogenesis. Ninety-second Meeting of the American Association for Cancer Research, New Orleans, LA, 2001. Su J.D., De, P. and Durden, D.L. PTEN controls matrix degradation and tumor invasion. Ninety-third Meeting of the American Association for Cancer Research, San Francisco, CA, 2002. Dey, N., De. P., Robertson, K.A., and Durden, D.L. Regulation of retinoic acid induced differentiation of neuroblastoma by SRC kinases. Advances in Neuroblastoma Research, Paris, France, 2002 Dey, N., Su, J.D., Epstein, A., Reynolds, C.P., Moritake, H., Sugimoto, T. and Durden, D.L. PTEN-AKT-MDM2-p53 axis in neuroblastoma therapeutics. Advances in Neuroblastoma Research, Paris, France, 2002. Quilliam, L. A., Yu, L., De, P., Yamuchi, A., Stone, J.C., Dinauer, M.C. and Durden D.L. Loss of RAP1A results in altered NADPH oxidase and haptotaxis in myeloid cells. American Society for Experimental Biology, 2003. Dmitry, T., De, P., March, K. and Durden, D.L. Role of Rac2 protein in restoration of blood perfusion in hind-limb ischemic mouse model. American College of Cardiology, 2003. Su, J. D., Garlich, J.R., Epstein, A., Tran, L.B., Mead, L.E., Ingram, D.A. and Durden, D.L. v integrin targeted PI-3 kinase inhibitors induce p53 transcription and apoptosis in endothelial cells. New agents in angiogenesis research, American Association for Cancer Research, 2003. De, P., Peng, Q., Dey, N., McDermitt, B., Peng, X., Garlich, J., Lonial, S., Durden, D.L. A vascular targeted pan PI-3 kinase inhibitor, SF1126 with activity against Multiple Myeloma in vivo. 48rd Annual Meeting American Society for Hematology (Oral Presentation) 2006.


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Joshi, S., Singh, A.R., Zulcic, M., and Durden, D.L. Role of Syk-Rac2 signaling axis in regulation of metastasis. AACR Symposium on Molecular Mechanisms of Cancer Metastasis, San Diego, CA, American Association for Cancer Research, 2012. Singh, Alok, Joshi, S. Garlich, J.R, Morales, G.A., and Durden D.L. Dual PI-3K/BRD4 inhibitors for maximal MYC control in cancer therapeutics. AACR Symposium on Targeting the PI-3 kinase-mTOR network in Cancer, Philadelphia, PA, American Association for Cancer Research, September 14-17, 2014. Recent Press Releases: “Hyundai Hope on Wheels awards Dr. Donald Durden $250,000 for cancer research” http://www.rchsd.org/about-us/newsroom/press-releases/hyundai-hope-on-wheels-awards-dr-donald-durden-250000-for-cancer-research/ September, 2013. “Novel therapeutic agent for pediatric cancer developed at UCSD in clinical trials; This is first time that a PI-3 kinase inhibitor has been used to treat a child with cancer” https://health.ucsd.edu/news/releases/Pages/2015-08-11-therapeutic-target-for-pediatric-cancer-devleoped-at-uc-san-diego.aspx September, 2015. Other press releases: http://www.dddmag.com/news/2015/08/novel-therapeutic-agent-pediatric-cancer-developed-clinical trials “SignalRx announces first pediatric patient with cancer treated with a PI-3 kinase inhibitor” http://www.firstwordpharma.com/node/1308314#axzz3nR8p1v9w http://www.ncbi.nlm.nih.gov/pubmed/25578041 R. EDUCATIONAL ACTIVITIES (1993-2012) Post-Doctoral Fellows: Hwang Min Kim, M.D. Dates: 6-93 to 6-94. Project: Role of hck and csk in FcRI and neuronal signal relay. Position: Assistant Professor Pediatrics, Yonsei University School of Medicine, Chunbuk, South Korea Ming Liu, M.D. Dates: 1-93 to 9-98 Project: Paracrine effects of neuroblastoma cell secreting cytokines on macrophage signal relay: Implications for tumor antigen presentation. Rae Kil Park, M.D., Ph.D. Dates: 5-94 to 5-97. Project: Role of the adapter protein-cbl interaction and raf-1 in FcRI signaling. Award: Children’s Hospital Career Development Fellowship. Position: Assistant Professor Microbiology and Immunology, Wongwang University, South Korea. Ivette Gonzales, M.D. Dates: 11-94 to 11-96. Project: Role of vav proto-oncogene and its interaction with adapter proteins in FcRI signal transduction.


26

Anat Erdreich-Epstein, M.D., Ph.D. Dates: 1-95 to 5-99. Project: Role of v3 integrin in tumor angiogenesis; a potential therapeutic target for treatment of pediatric neural tumors. Mechanisms of integrin and ITAM signaling in endothelial cells. Awards: Childrens Cancer Research Foundation, Concern II foundation, NIH RO1. Position: Associate Professor, Tenured, USC School of Medicine, Los Angeles, CA Wade T. Kyono, M.D. Dates: 7-95 to 8-97. Project: Role of ckl and other adapter proteins in FcRI and FcRII signal relay. Position: Assistant Professor Pediatrics, University of Hawai. Honolulu, HA Julie Chu, M.D. Dates: 7-96 to 7-98 Project: Role of SLP-76-CBL-GRB2-SHC adapter interaction in regulation of RAS. Position: Assistant Professor Pediatrics, Oregon Health Sciences Center, Portland, Oregon Anita Kant, PhD. Dates 8-97 to 5-99 Project: Role of CRKL, HEF-1 adapter proteins and Phosphatases regulation of myeloid signaling. Award: CHLA Career Development Fellowship. “Phagocytosis in Macrophages: Signal transduction through Fc Receptors”, 1998 Andreas Reiff, M.D. Dates: 7-97 to 5-99 Project: Treatment of collagen-induced arthritis with L-asparaginase: A Preclinical Study. Award: CHLA Junior Faculty Career Development Award, 1998. Arthritis Foundation Grant, 1998 Position: Professor, USC School of Medicine Nandini-Rudra Ganguly, Ph.D Date: 9-98 to 5-99 Project: Endothelial TrkA signaling. Position: Baxter Pharmaceuticals, Los Angeles, CA. Shenghua Wen, M.D., Ph.D. Date: 7-99 to 8-2000 Project: Role of PTEN in control of tumor-induced angiogenesis Position: Staff Scientist, Millennium Pharmaceuticals, Cambridge, MA. Jong-Suk Kim, M.D., Ph.D. Date: 11-98 to 6-03 Project: Role of SHP-1, SHIP and PTEN phosphatase in FcR signaling. Professor, Chonbuk National University, Korea. Amy Marie Munchhof Date: 6-2001 to 12-2003 MD/PhD Student Role of PTEN in angiogenesis and tumor progression. Kamnesh Pradhan MD Date: 7-2001 to 7-2003


27

MD/Hematology/Oncology Fellow Role of PTEN and PI-3 kinase-AKT-MDM2-p53 pathway in chemo-radiation sensitivity. Rapamycin/VP-16 clinical trial to target AKT/mTOR pathway in solid tumors. Position: Assistant Professor, Dept. of Pediatrics, Riley Children’s Hospital, Indianapolis, IN. Pradip De, Ph.D. Instructor, Faculty member, Emory University School of Medicine Date: 7-99 to 2008 Project: RAC2 knockout model for myeloid signaling. Xiaodong Peng, Ph.D. Date: 8-99 to 12-2003 Role of Cbl and other adapter proteins in myeloid Fc receptor signaling. Position: Staff Scientist, Semafore Pharmaceuticals, Indianapolis, IN. Nandini Dey, Ph.D. Instructor, Faculty member, Emory University Date: 11-99 to 2008 Project: Role of Src, Csk and TrkA in neuronal proliferation and differentiation. Jing Dong Su, MD. Ph.D. Date: 7-2000 to 12-2003 Project: Role of PTEN in control of tumor induced angiogenesis, invasion and metastasis Position: Senior Research Scientist, Semafore Pharmaceuticals, Indianapolis, IN. Hal Crosswell, MD Date: 7-2004 to 6-2006 MD/Hematology/Oncology Fellow Role of PTEN in glioma and RMS transformation Assistant Professor, University of South Carolina, Greenville, SC Gabriela D C Denning, PhD Date: 8-2004 to 8-2007 Post Doctoral Fellow PTEN localization controls tumor suppressor function of PTEN ACS Postdoctoral fellowship recipient 2004- 2006 Qiong Peng, PhD Date: 7-2005 to 2009 Post Doctoral Fellow PTEN genetics: partners for PTEN function as tumor suppressor Recent and Current Members of Durden laboratory/Projects: Robert Craig Castellino, MD Date: 1-2007 to 2009 Assistant Professor, Dept of Pediatrics, Emory University PTEN and SHH/PTCH/SMO signaling in medulloblastoma biology Arshad Ahsanuddin, MD Date: 1-2007 to 2008


28

Post Doctoral Fellow, Dept of Pathology PTEN and PI-3 kinase in hematopoeitic malignancy, multiple myeloma and lymphoma therapeutics Carrie R. Muh, MD Date: 1-2008 to 2009 Post Doctoral Fellow, Dept of Neurosurgery, Emory University PTEN and PI-3 kinase and Rac2 in brain tumor angiogenesis and therapeutics Susan Peirce, PhD Date: 10-2008 to 2009 Post Doctoral Fellow, Dept of Pediatrics, Emory University PTEN and PI-3 kinase and Rac2 in lymphomagenesis and neuroblastoma progression and therapeutics. Lisa Hartman, MD Date: 10-2009 to 7-2012 Post Doctoral Fellow, Dept of Pediatrics, UCSD PI-3 kinase inhibitors for the treatment of embryonal pediatric tumors, polyICLC Phase II trial in LGG Mohar Chattopadhyay, PhD Date: 11-2010 to 10-2011 Post Doctoral Fellow, Dept of Pediatrics, UCSD PTEN and PI-3 kinase and Rac2 in medulloblastoma cancer stem cell biology Andrew Martin, MD Date: 7-2012 to 6-2014 Post Doctoral Fellow, Dept of Pediatrics, UCSD PTEN and integrin signaling in medulloblastoma cancer stem cell biology Alok Singh, PhD Date: 10-2009 to present Project Scientist, Dept of Pediatrics, UCSD PTEN and PI-3 kinase in medulloblastoma progression and cancer stem cell biology Shweta Joshi, PhD Date: 10-2009 to present Project Scientist, Dept of Pediatrics, UCSD PTEN and PI-3 kinase and Rac2-Syk in the control of M1-M2 macrophage differentiation, angiogenesis, tumor growth and metastasis Amanda Goldin, MD Date: 7-2015 to present Post Doctoral Fellow, Dept of Pediatrics, UCSD Role of PI-3 kinase in Ewings sarcoma therapeutics Francisco Vega, PhD Date: 6-2015 to present Visiting Faculty, Dept of Pediatrics, UCSD Role of Rac2 and Vav1 in adaptive immunity and metastasis. Dual PI-3K/PARP inhibitor of neuroblastoma therapeutics, synthetic lethality on DNA repair pathways. Adam Burgoyne, MD, PhD


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Date: 7-2015 to present Post Doctoral Fellow, Dept of Pediatrics, UCSD PI-3 K and epigenetics in hepatocellular carcinoma and GIST Graduate and Undergraduate Students: Michael Alcaraz (Undergraduate, UCSD) Date: 7-2013 to present Role of PTEN in CSC pathology in medulloblastoma Cassandra Wang (Undergraduate, UCSD) Date: 2009 to 2013 Asparaginase in leukemia Current: Medical School, USC School of Medicine Rami Banitji (Undergraduate, Williams College, Premedical) Summer 1993: “Role of trkA and csk in neuronal differentiation” Summer 1994: CHLA Oncology Fellowship, “Role of hck and vav in FcRI signal transduction” Steven Yoon (Medical Student, New Jersey, Medical School) Summer 1995: CHLA Summer Oncology Fellowship Project: Role of the syk protein kinase in FcRI phagocytosis Scott D. Miller (Medical Student, University of Iowa College of Medicine) Summer 1995: CHLA Oncology Fellowship Project: Role of small GTPase of the ras and rac family in mammalian programmed cell death. Kayvon D. Izadi (Creighton University School of Medicine ) CHLA Oncology Fellowship Dates: 12-96 to 7-1999. Project: Role of NCK-Cbl adapter protein interaction and PAK kinase in FcR signaling in myeloid cells. Graduate Student Committees: Steven Mullen (PhD, Purdue University) Cryopreservation related stress signaling in embryos” Marianne Price (PhD, Dept. of Genetics, IU School of Medicine) “Mechanisms for regulation of respiratory burst response in myeloid cells” Amy Marie Munchhof (MD, PhD, Indiana University School of Medicine)(Chairman) “PTEN in the control of tumor progression, mechanisms of regulation” Invited Lectures (National and International) 01/01/93 “A role for Syk and Shc in FcRI signal transduction”, Division of Research Immunology

Seminar Series, Childrens Hospital Los Angeles (CHLA), Los Angeles, CA)

02/10/93 “Mechanism of signaling through the FcRI receptor”, Department of Pathology Lecture Series, CHLA, Los Angeles, CA.

03/10/93 “MDR molecular basis for drug resistance", Division of Hematology and Oncology


30

Seminar Series, CHLA, Los Angeles, CA.

03/10/93 “Syk or Shc, take your pick: FcRI signal relay”, Tumor Biology Group, Norris Cancer Center, USC School of Medicine, Los Angeles, CA.

12/10/93 “Chimeric receptor molecules designed from the antigen receptor homology one domain

motif”, Gene Therapy Meeting, CHLA and Norris Cancer Center, USC School of Medicine, Los Angeles, CA.

05/09/94 “Paracrine effects of cytokine secreting neuroblastoma cells on macrophage signal

transduction”, Gene Therapy Retreat, CHLA and Norris Cancer Center, USC School of Medicine, Los Angeles, CA.

06/10/94 “Syk and Shc in FcRI signal transduction”, Department of Immunology, Scripps Clinic and

Research Foundation, La Jolla, CA.

03/10/95 “Fundamental feature of mammalian signal transduction: oncogenes and proto-oncogenes”, CHLA, Los Angeles, CA

04/15/96 “Signal transduction mechanisms controlling RAS in myeloid cells”, University of Iowa

College of Medicine, Iowa City, Iowa.

06/19/96 “Grb2-SH3-SH2-cbl interaction in immunoreceptor tyrosine activation motif (ITAM) signaling”, Twelfth Annual Meeting on Oncogenes, Frederick, Maryland, Presentation

10/01/96 “Mechanisms of myeloid cell signaling”, Wonkwang University School of Medicine, Korea.

10/02/96 “Molecular basis for Fc receptor signaling in myeloid cells”, Symposium of Cell Signal

Transduction (Keynote Speaker), Institute for Cardiovascular Research Chonbuk National University,Chonbuk, Korea.

10/04/96 “A role for SHC, GRB2 and Raf-1 in macrophage signaling”, Plenary session, Korean

Society for Immunology, Taejon, Korea,

10/07/96 “Fundamental Mechanisms of mammalian signal relay”, Yonsei University Wonju College of Medicine, Korea.

11/12/96 “FcRI signaling: A paradigm for adapter protein interactions in the regulation of Ras, CHLA Research Institute, Los Angeles, CA.

01/26/97 “General features on mammalian signal transduction FcRI signaling as a model”, Cell

Matrix Group, CHLA, USC School of Medicine, Los Angeles, CA.

05/10/97 “BCR/ABL transformation: molecular mechanisms”, Pediatric Faculty Meeting, CHLA, Los Angeles, CA.

10/14/97 “Angiogenesis and neuroblastoma”, Childrens Cancer Group Meeting, Neuroblastoma

Strategy Group, San Diego, CA.

11/18/97 “FcRI signaling in myeloid cells; a paradigm for complex adapter protein regulation of Ras, University of Texas, Southwestern University, Dallas, Texas.


31

12/18/97 “Role of vascular integrins v3 and v5 in angiogenesis”, Cell Matrix Group, CHLA,

Los Angeles, CA.

01/15/98 “Recombinant Wolinella asparaginase for the treatment of leukemia” Childrens Hospital Los Angeles, International Symposium on Asparaginase Pharmacology and its implications in the treatment of leukemia and lymphomas, Los Angeles, CA.

02/06/98 “Control of cellular functions by the extracellular matrixUSC Cancer Center

Minisymposium CHLA, Los Angeles, CA.

03/11/98 “Signal transduction” Division of Infectious Diseases, CHLA, Los Angeles, CA.

05/03/98 “Anti-angiogenic therapy for the treatment of cancer”, Pediatric Faculty/Research Faculty Meeting, CHLA, Los Angeles, CA.

05/15/98 “Wolinella asparaginase in the treatment of acute lymphoblastic leukemia”, Martell Site

Visit, CHLA, Los Angeles, CA.

07/28/98 “FcR signaling: A paradigm for study of complex adapter proteins in the regulation of RAS”, Combined Seminar, Indiana University School of Medicine, Indianapolis, IN.

11/01/99 “Myeloid Immunoreceptor Signaling: A Paradigm for Study of Receptor Modulation of Small GTPases in Mammalian Cells”, Biochemistry Department, Indiana University School of Medicine, Indianapolis, IN.

08/13/00 “PTEN controls the tumor-induced angiogenic response”, Tyrosine Phosphorylation and

Cell Signaling Meeting, The Salk Institute, La Jolla, CA. 06/10/00 “Myeloid Immunoreceptor Signaling: A Paradigm for Study of Receptor Modulation of Small

GTPases in Mammalian Cells”, Indiana University School of Medicine. 04/22/00 “Two Vignettes for integrin function and phosphatase action in angiogenesis control” Vascular Biology Retreat, Indianapolis, IN 07/13/00 “PTEN Controls the Growth and Angiogenic Response of Malignant Gliomas”, Tyrosine Phosphorylation and Cell Signaling Meeting, The Salk Institute, La Jolla, CA. 08/22/00 “PTEN controls angiogenesis and inflammatory signaling” Department of Microbiology and Immunology, Indiana University School of Medicine. 01/12/01 “Signals that control angiogenesis” Pediatric Faculty Seminar, Department of Pediatrics. 05/12/01 “PTEN is the angiogenic Switch” American Cancer Society Midwest Investigators Meeting, Indianapolis, IN 07/21/01 “PTEN controls angiogenesis and inflammatory signals” Millennium Pharmaceuticals, Boston, MA. 08/12/01 “PTEN: Angiogenesis and Inflammatory Signaling” Department of Immunology, Scripps Clinic

and Research Foundation, La Jolla, CA.


32

12/13/01 “PTEN controls tumor-induced angiogenesis and tumor progression through modulation of p53” Cancer Biology Seminar Series, Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA.

02/09/02 “Angiogenesis and Neuroblastoma therapeutics” Annual Neuroblastoma NANT/PPG Meeting, Los Angeles, CA

06/12/02 “PTEN/AKT/p53 axis in Neuroblastoma progression” Advances in Neuroblastoma Research, Paris, France (Plenary Presentation). 09/11/02 “PTEN and tumor progression via the through modulation of p53” University of Cincinnati School of Medicine, Cincinnati, OH (Hem/Onc Rounds)

10/18/02 “PTEN/AKT/p53 signaling axis in glioma therapeutics” Grand Rounds Childrens Hospital Los Angeles, USC School of Medicine Los Angeles, CA 12/03/02 “PTEN is the angiogenic switch in tumor progression” Lineberger Cancer Center Grand Rounds, University North Carolina, Chapel Hill, NC. 2/11/03 “Integrin Signaling in Neuroblastoma Therapeutics” NANT Annual Meeting, Los Angeles, CA. 2/22/03 “PTEN/PI-3 kinase as therapeutic target for cancer” Emory University School of Medicine, AFLAC Visiting Professorship, Atlanta, GA. 4/11/03 “PTEN-RAC2 axis in angiogenesis” Vascular Biology Meeting, Indiana University School of Medicine, Indianapolis, IN. 6/23/03 “Wolinella asparaginase for treatment of acute leukemia” RAID NCI Meeting Developmental therapeutics, Frederick, MD. 4/2/04 “PTEN inhibitors for cell protection” Department of Energy, Symposium of Inositols in Cell Protection, MIT, Cambridge, MA 8/9/04 “PTEN and PI-3 kinase inhibitors in Cancer Therapeutics” Keynote address: Molecular Targets in Cancer Therapeutics, Mayflower Hotel, Washington, DC. 5/26/05 “PTEN and PI-3 kinase in Cancer therapeutics” Medical University of South Carolina, Hollings Cancer Center Seminar, Charleston, SC. 8/12/05 “Update on PI-3 kinase inhibitors for Cancer Therapy” Molecular Targets for Cancer Therapy, Washington, DC. 9/21/05 “PTEN and PI-3 kinase inhibitors in Cancer Therapeutics” Cambridge Biotech Meeting. Cambridge, MA. 8/26/06 “PTEN and PI-3 kinase inhibitors for Cancer Therapy” 5th Annual Molecular Targets for Cancer

Therapy, New York, NY. 9/12/06 “Angiogenesis: PTEN and PI-3 kinase the Yin and Yang for Angiogenesis” Vascular Biology

Program Seminar, Cornell Weill Medical Center, New York, NY. 12/12/06 “Vascular Targeted pan PI-3 kinase inhibitor, SF1126 for Multiple Myeloma” 48rd Annual ASH

meeting, Orlando, FL.


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9/24/07 “Emergence of PI-3 kinase as a validated target for cancer therapy” Plenary introduction to: PI-3 kinase workshop. 2007 Fall CTEP early drug development meeting, Bethesda, MD. 12/5/08 “PTEN-PI-3K-RAC2 axis: Determinant of specificity or signaling and target for therapeutics” Special Seminar in Cancer Biology, UCSD Moores Cancer Center, San Diego, CA. 3/25/09 “The molecular biology of the PI-3 kinase/AKT/mTOR signaling pathway” 7th International Symposium on Targeted Anticancer Therapies, March 23-25, 2009 Amsterdam, The Netherlands. 8/25/10 “PI-3 kinase inhibitor for anticancer therapeutics”9th Annual Symposium on Targeted Anticancer Therapies, August 23-25, 2010, Washington, DC, USA. 11/12/12 “Signal transduction therapeutics for cancer and pediatric disease” Pediatric Grant Rounds,

UCSD Rady Children’s Hospital, San Diego, CA. 5/13/13 “Targeted therapy in pediatric oncology” Greehey Institute Visiting Professorship, University

of San Antonio, Department of Pediatrics, San Antonio, TX 9/25/13 “Stromal factors that are targets for PI-3kinase inhibitor therapeutics in the control of

metastasis” Seventh Annual, Novel Strategies for Kinase Inhibitors, Discovery on Target, Boston, MA, USC.

4/15/15 “Dual PI-3 kinase/epigenetic inhibitors for cancer therapeutics” 9th Annual Discovery on

Target, San Diego, CA. 9/22/15 “Dual PI-3 kinase/ BET bromodomain inhibitors for therapeutics” 10th Annual Discovery on

Target, Boston, MA. Epigenetic targeted therapy and chromatin modification. 4/19/16 “Dual PI-3 kinase/ BET bromodomain inhibitors for therapeutics” 11th Annual Discovery on

Target, San Diego, CA. Epigenetic targeted therapy and chromatin modification.


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S. RESEARCH INTERESTS & PROGRAMATIC VISION FOR DEPT OF PEDIATRICS We provide URL for immediate access to the all 67 publications from Durden laboratory and UCSD Durden laboratory and SignalRx Pharmaceuticals web sites: 23 MS since 2009.

http://durdenlab.ucsd.edu http://www.ncbi.nlm.nih.gov/pubmed/?term=durden+dl http://www.signalrx.com/ My laboratory is interested in understanding the fundamental mechanisms by which signals are transmitted from cell surface receptors to the cytoplasm and nucleus. Our efforts are focused on the role of lipid and protein phosphorylation and dephosphorylation in the regulation of signal transduction and epigenetic effector mechanisms. In this regard, I am primarily interested in the regulation of signal transduction pathways in mammalian cells which might encode pathophysiologic states in pediatrics. We are interested in converting target discovery and validation into drug discovery and drug development in particular as it relates to specific kinases and/or epigenetic effectors and translating this basic science information from "bench to bedside” in pediatric medicine. My overarching interest has been, over the past 20 years, to create a collaborative academic environment in which “Scientific Discovery” identifies a precision targeted therapeutic for refractory pediatric disease states. My scientific training is not just in pediatric hematology and oncology but in the dissection of mammalian/human signaling pathways for therapeutic gain. This is one reason I was recruited to serve as the Vice Chair for Research at UCSD in 2009. My academic leadership focus over the past 6 years as Vice Chair for Research in the Department of Pediatrics at UCSD/Rady Children’s Hospital has been to the establish an infrastructure which supports outstanding patient care while as same time augmenting clinical and basic research within the Department of Pediatrics. One component of this was to establish a sophisticated research infrastructure to empower and stimulate all clinical and basic faculty to perform translational research. The focus was to build an effective mechanism to integrate basic and translational research within the Rady/UCSD academic environment. A number of strategies were employed to include: 1) development of a robust biorepository system with frequent outreach to all clinical investigators in the system 2) to present 30-40 talks a year to all Divisions and Departments within the UCSD/Rady clinical umbrella opportunities to interface with this infrastructure for translational research. The results were remarkable; we successfully initiated within the first year over 30 new translational research projects at UCSD/Rady Children’s Hospital in 10 different Divisions and/or Departments all devoted to attacking an important pediatric unmet medical needs. I have personally championed this effort and made many connections between the UCSD/Rady clinical investigators and researchers within the UCSD research mesa and Industry (e.g. Salk Institute, Sanford-Burnham, Scripps/TSRI, etc.).The Durden laboratory has been involved in small molecule drug discovery and development in the area of PI-3 kinase inhibitors for the last 16 years. I am focused on the dissection of the molecular basis of signaling pathways in refractory problematic pediatric diseases in the context of drug discovery and drug development. In this regard, my laboratory developed the first PI-3 kinase inhibitor, SF1126 to enter pediatric oncology clinical trials in 2015 in the treatment of recurrent and/or refractory Neuroblastoma.


35

Finally, as a board certified practicing attending pediatric hematologist-oncologist at UCSD, I am actively involved in the development and execution of Phase I and Phase II clinical trials of targeted and immunotherapies in cancer patients at the UCSD/Rady Children’s Hospital and Moores Cancer Center and participation in our weekly molecular tumor boards. I understand the potential challenges to drug development in the Pediatric population including but not restricted to cancer. This element of my training, I believe gives me an additional level of insights into how to build an academic program which studies and eventually applies targeted therapeutic agents in a genome driven/precision medicine manner to refractory pediatric disease states. I. “PI-3 kinase and PTEN signaling in mammalian systems” i. PTEN and PI-3 kinase signaling pathway as central nodes in the control tumor-induced angiogenesis. In 2001, we reported that the tumor suppressor PTEN controls tumor-induced angiogenesis (PNAS: 98: 4622, 2001). PTEN is a dual specificity lipid/protein phosphatase now implicated as a tumor suppressor gene in a significant number of human cancers including brain tumors (30-40%), endometrial carcinoma (90%), prostate cancer (40%), breast cancer, lymphomas, etc. Mutations in PTEN correlate with more invasive angiogenic phenotype leading us to hypothesize that PTEN may be important in tumor progression. We used an orthotopic brain tumor model to investigate the role of PTEN in brain tumor progression. Glioblastoma cell lines (U87MG, U373MG) which are null for PTEN were genetically reconstituted with PTEN or missense mutants of PTEN in order to determine if PTEN controls tumor progression in vivo. Our results demonstrate the first direct evidence that PTEN controls the tumor-induced angiogenic response in this model system. More importantly, this work lead to the formulation of the “intercept node” hypothesis (Castellino and Durden, Nat.Clin. Practice Neurology 3:682,2007) for target discovery/identification. We and others have shown that PTEN exerts a control over p53 transcription and many other downstream networks within the tumor and stromal cell through the phosphorylation of AKT and the control of MDM2 (J. Biol. Chem. 77: 777, 2002; Cancer Res.63: 3585, 2003). Recently we defined a novel role for AKT and MDM2 in the cytoplasmic regulation of the hypoxic stability of HIF1 (J Biol. Chem., 2013).

This novel control circuit would coordinate proliferation and the induction of angiogenesis. We propose a model where PTEN and p53 exert a coordinate control over a large number of signaling events (Figure 1). In normal cells, this switch mechanism is highly regulated whereas during tumorigenesis with loss of tumor suppressor function i.e. PTEN, p53, etc. the control over proliferation, apoptosis and angiogenesis is lost

Fig. 1. PTEN/PI-3K exerts coordinate “central nodal” control over a large number of mammalian signaling events. Hence it could be viewed as “master control switch” for proliferation, angiogenesis, migration, differentiation and metastasis. The concept of coordinated intracellular regulation of angiogenesis is proposed as a model. Hence the PI-3 kinase signaling axis is viewed as a central element of mammalian signaling. Both these pathways are mutated at high frequency in human neoplasia. These pathways are major targets for anticancer therapeutics. In collaboration with Semafore Pharmaceuticals we have successfully developed both PI-3 kinase and PTEN inhibitors to control the “Yin and Yang” of mammalian signaling for therapeutic gain. Castellino and Durden, Nat.Clin. Practice Neurology 3:682,2007.


36

leading to uncontrolled growth, induction of vascular proliferation and metastasis. In other experiments we have shown that PTEN controls tumor cell migration, invasion and matrix degradation. Moreover PTEN contributes to chemoradioinsensitive state of tumor cells via its control of p53 survival and DNA repair signaling pathways. We have developed small molecule PTEN inhibitors in silico. ii. Drug Discovery and development; Targeted therapeutics using RGD-integrin targeted PI-3 kinase inhibitors and PTEN inhibitors; Discovery and Development of SF1126 “Bench to bedside”. It is now clear that a greater understanding the PTEN and PI-3 kinase signaling networks and the downstream targets of this tumor suppressor-oncogene pair will identify new therapeutic targets for clinical intervention since this pathway is mutated and/or altered at high frequency in human tumors. This area of investigation was funded by the NIH RO1 CA94233-09.We have now generated v-targeted pan-PI-3 kinase inhibitors as potential antiangiogenic agents for cancer therapy (SignalRx Pharmaceuticals). Based on these studies we have developed an RGD integrin targeted small molecule inhibitor, SF1126 for clinical application (Cancer Research 68: 206, 2008) (structure shown below). It was one of the first pan PI-3 kinase inhibitors to enter human clinical trials in cancer. We have now executed and completed the Phase I trial of SF1126 (Eur. J. Cancer, 2012). In 2015, we executed the first Phase I trial of a PI-3 kinase inhibitor in pediatric oncology via the NANT consortium and a Phase II genome driven bucket trial in adult cancers with PIK3CA mutations. This was first time a child was treated with this class of a therapeutic agent. Dual inhibitory chemotype discovery. We have now gone on to develop in silico a large number of dual targeted small molecules in a pipeline for the treatment of pediatric diseases. Currently, we have a large in silico drug discovery effort fueled by X-ray crystallography/molecular modeling and built around the PI-3K signaling pathway. We have hundreds of small molecules in our pipeline which inhibit these targets at nM potency with good safety profiles in animal models for toxicity (J Med. Chem 2012). Dual inhibitory chemotypes developed so far include: 1) PI3K/MEK 2) PI-3K/PARP 3) PI-3K/BRD4 (SF2523 (Fig 2) 4) PI-3K/HDAC 5) PI3K/CDK4/6 and others. We have solved the co-crystallized structure of our dual PI-3K/BRD4 inhibitor, SF2523, bound to the active site of BRD4 at 1.8 Angstroms (BD1) (Fig. 2). Fig. 2. Dual inhibitor of PI-3K and BRD4, SF2523. We show the co-crystal of BRD4 protein (BD1) and SF2523 (magenta) or JQ1 (grey color) solved at 1.8A for the binding of SF2523 to the BRD4 binding domain 1 (BD1). Grey regions are hydrophobic, red regions are negatively charged and blue areas are positively charged domains

of BRD4/BD1. This chemotype is one of many dual inhibitory small molecules developed in the Durden

laboratory for different therapeutic areas based on synthetic lethalities or synergistic oncogene interactions (listed above).

O

O

N+

O

O

O

O

N

N

N

O

N

O

N

O

O

N+

O

N

OH

O

O

H

H

HH

R

N

N

NH3+

O

N

O

N

O

O

N+

O

N

OH

O

O

H

H

HH

R

O

O

N

O

O

O

N+

O

O

O

OR1

SF1126 R= -HSF1326 R= -CH3

SF1101 (LY294002)

"RGDS" R= -H"RADS" R= -CH3 SF1110 R=-O-(t-Bu)

SF1111 R= -Cl

O

O

S

N

O

O O SF2523


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iii. Targeted therapeutics against kinases, phosphatases and epigenetic effectors e.g. PTEN, PI-3 kinase and BRD4.

Drug Discovery efforts focused on PI-3 kinase or phosphatases is a novel area of investigation in the Durden laboratory. In collaboration with the chemists at SignalRx Pharmaceuticals we have developed small molecule inhibitors of PI-3 kinase and the PTEN phosphatase. Using recombinant PTEN or mutants of PTEN we have screened for inhibitors of this important intercept node. Our idea is that a coordinated control of phosphatase/kinase pairs will yield a condition of “signal manipulation” or therapeutic gain. We have used a number of strategies for drug discovery and development built around our interest in signaling pathways which serve as major intercept components of pathophysiologic states. These drug targets include: 1) PTEN 2) PI-3 kinase 3) Syk and 4) Csk kinase and 5) epigenetic effectors e.g. BRD4. Multiple methods are used including: 1) in silico pdb based structural modeling and screens 2) in silico ab initio modeling algorithms 3) high throughput and high content screens of multiple libraries and 4) chemoinformatics 5) NMR and X-ray crystal structural analysis of target-drug interface. In conjunction with this area of investigation, I cofounded a small molecule drug discovery company, SignalRx pharmaceuticals, to develop inhibitors of the PI-3 kinase pathway and more recently dual inhibitory chemotypes for pediatric targeted therapeutics. We have developed the “first in class” pan PI-3 kinase and PTEN small molecule inhibitors (Garlich et al, Cancer Res, 2008). We have now obtained FDA approval for our PI-3 kinase inhibitor and have completed the adult Phase I clinical trials of this agent (Eur. J. Cancer, 2012). We have now executed the first pediatric Phase I trial for this class of agent: “This is the first time a child with cancer was treated with a PI-3 kinase inhibitor”. https://www.clinicaltrials.gov/ct2/show/NCT02337309?term=SF1126&rank=2 iii. Tumor microenvironment (TME) signaling pathway controlling tumor progression and metastasis via the provisional integrin signaling network. Below we show results which highlight are recent discovery in Durden laboratory, the identification of a signal transduction pathway in macrophages which extends from the provisional integrin, 41 through a kinase/GTPase pair (Syk-Rac2) to control tumor metastasis in vivo. This signaling network has been shown to control macrophage M1-M2 differentiation, tumor progression and metastasis in vivo (PLoS One, 2013). RNA sequencing and CHIPseq have defined an entire transcriptomic program for the control of macrophage M1 to M2 transition and metastasis.

Intercept Hypothesis

PTENPI-3 kinase

AKT/PKB

Growth factors

Nuclear compartment

LY294002/SF1126

mTOR other targets

p53

mdm2

“INTERCEPTNODE”

Fig. 3. The intercept concept can be summarized as: a point in mammalian signaling where multiple cell surface receptors converge. We have attacked a major intercept point in mammalian signaling by developing PTEN phosphatase inhibitors. These we obtained from in silico screens for small molecules which would interact with the P loop structure within the crystal structure coordinates of PTEN as shown.


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iv. Signaling pathways in cancer stem cells (CSCs) in medulloblastomagenesis. In 2009, we engaged in the study of cancer stem cells (CSCs) in a well-developed Smo transgenic mouse model (Castellino et al, PLoS One, 2010). We have now defined the fundamental molecular features of the CSC (“stemness signaling properties”) which encode sensitivity and resistance to targeted therapeutic agents including PI-3K inhibitors vs chemotherapeutic drugs. These observations will have therapeutic implications for the treatment of cancers which are stem cell driven. II. Molecular immunology: Molecular basis for Fc receptor signaling in myeloid cells: Targets for drug discovery to control ITIM and ITAM motifs for therapeutic gain? The Durden laboratory has maintained its interest in molecular immunology since 1992 and continued to search for new immunomodulatory targets for immuno-therapeutic exploitation. Most recently in 2013, we reported evidence that the phosphatases SHP1 and PKC regulate the FcR signaling axis (ECR, 2013). Evidence from several knockout mouse models has demonstrated conclusively that the Fc receptors are critical elements in the control of autoimmunity and inflammatory diseases. The FcRs are involved in both positive and negative regulation of immune responses and inflammatory cascades. My research primarily involves the study of how tyrosine phosphorylation and dephosphorylation of cell proteins and lipids drives myeloid signal relay pathways. Over the past 12 years, my laboratory has developed a model for how the Fc receptor for IgG transmits intracytoplasmic signals to the respiratory burst and other pathways. (see schematic below). An attraction to this area of research is the clinical need to learn more about neutrophils and the activation of other myeloid cells (platelets, macrophages, dendritic cells, mast cells, etc.) which impact on a number of human disease states (inflammatory diseases, coronary artery disease, autoimmunity, vaccine development strategies, immunotherapy for cancer, etc.). Recent work from knockout animal models has clearly demonstrated the role of Fc receptors in the control of inflammation and autoimmunity. The signaling pathways which control Fc receptor functions are critical targets for development of anti-inflammatory drugs and drugs to control autoimmunity, GVHD and graft failure. More recently we have begun to use a genetic approach in mice deficient in FcR signaling components to identify signaling pathways important for inflammatory responses in vivo. Our studies of FcRI as a model system has begun to contribute novel information to the understanding of myeloid signal relay. Below are described our current research interests and active areas of investigation relating to the study of the Fc receptor signaling pathways in myeloid cells. Our principal focus is to identify new drug targets for drug discovery platforms in an effort to develop agents which will augment or suppress ITAM or ITIM signaling networks for therapeutic gain. We will translate these discoveries from Bench to Bedside. i. Role of protein tyrosine phosphorylation and dephosphorylation in FcR/ITIM/ITAM signaling. Protein tyrosine phosphorylation and dephosphorylation are important in the regulation of hematopoietic signal transduction. Protein tyrosine kinases (PTKs) of the receptor and nonreceptor class are implicated in myeloid signal relay and the protein tyrosine phosphatase, CD45, is clearly involved in the regulation of cell-cell signaling in T cells, B cells and myeloid cells. Inside the cell, PTKs and protein tyrosine phosphatases likely interact to control signaling through specific receptors by establishing protein-protein interactions mediated through SH3, SH2 and other domains, etc. I am interested in understanding how phosphatases, kinases and their associated cosignaling molecules might function in the coordination of normal signal relay pathways. Recently there is more interest the study of kinase/phosphatase pairs which provide critical rheostatic control of key signaling pathways in myeloid and endothelial cells.

RAC2 controls metastasis in vivo. We have elucidated a M2 macrophage autonomous signaling pathway that regulates tumor metastasis. The B16L10 melanoma metastasis model is shown. Upper panel metastatic nodules (black) in wild type mice, lower panel 5 mouse lungs from Rac2-/- animal (no metastasis detected).


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To approach this question, I have been using human myeloid cell lines to study signaling through the Fc receptor for IgG in part because several downstream signals are easily and rapidly measured in these cells. Our laboratory has now established that the Fc receptors for IgG signal through the tyrosine phosphorylation of cell proteins resulting in the rapid activation of an intracellular NADPH-oxidase complex. The latter response is termed the respiratory burst (RB). The molecules involved in the activation of the oxidase complex, including the small GTPases rap1a and rac, have been cloned and characterized and the NADPH-oxidase activity can be measured in crude membrane preparations and in a cell-free system. There are several advantages to the study of FcRI to the RB as a model system: 1) the assembly of the RB machinery is very specific, 2) it is easily measured both in vivo and in vitro using various combinations of recombinant proteins, 3) the RB is a rapid response occurring in 1-2 min therefore alteration in the response can be directly attributed to protein-protein interactions and not secondary to transcriptional activation of genes. 4) In vitro plasma membrane preparations allow for addition of recombinant molecules (dominant negatives or dominant positive signaling molecules) to ask questions regarding the regulation of FcRI signaling to the RB in vitro. We reported in a Rap1a knockout mouse model, a role for rap1a in macrophage integrin and FcR signaling (J Immunology, 179, 2007) and a role for SHP1 in regulation of phagocytosis via the FcgR (ECR, 2013). FcRI stimulation in myeloid cells is associated with the activation of specific myeloid nonreceptor protein tyrosine kinase and the tyrosine phosphorylation of a specific amino acid sequence contained within the FcRI subunit, termed the immunoreceptor tyrosine-based activation motif (ITAM) motif (consensus, YXXLX6YXXL). We study Fc receptor signaling using interferon differentiated U937 myeloid cells, termed U937IF cells. U937IF cells are incubated with monoclonal antibodies specific for FcRI or FcRII on ice for 30 min followed by addition of crosslinking antibody (rabbit anti-mouse, Fab’2 fragment). This is a very specific and potent method for cell surface aggregation of the FcR subunits and results in the activation of intracytoplasmic effectors of signaling that are linked to these subunits. We reported that FcRI stimulation results in the tyrosine, serine and threonine phosphorylation of the FcRI subunit (Biochem. J. 299:569, 1994; Exp Cell Res.211:150, 1994). We have identified the nonreceptor protein tyrosine kinases, hck, syk and Btk as components of FcRI signaling in myeloid cells (Blood, 84:2102, 1994; J. of Immunol. 154:4039, 1995, Blood, 89:388, 1997). It is important to identify and carefully characterize the upstream events that occur immediately after receptor activation as a prerequisite for a detailed analysis of the downstream signaling events. Critical questions remain to be answered. In particular, what are the key substrates for upstream tyrosine kinases and protein tyrosine phosphatases that transmit different signals through FcRI i.e activation of the RB, (transcriptional activation of cytokine genes, antibody-dependent cell-mediated cytotoxicity, phagocytosis, motility responses, etc.)? This information will likely impact on the development of immunopharmacologic agents to control macrophage, neutrophil and mast cell activation for the treatment of human disease including cancer via immunotherapeutics. ii. Role of complex adapter proteins in FcRI, ITIM and ITAM signaling. The function of tyrosine phosphorylation as an initial event is to phosphorylate specific substrates which will then mediate the downstream activation of serine/threonine kinases and other kinases to generate phospholipid metabolites and other secondary messengers. The initial substrates for tyrosine phosphorylation are known as complex scaffolding or adapter proteins and are important for the encoding of the specificity of signaling (Blood, 92: 1697, 1998). We have been very interested in the study of the Cbl adapter protein (J. Immun. 160: 5018, 1998). Recent evidence from Cbl knockout mice suggests that Cbl may control the induction of autoimmunity. Cbl is known to bind to a large number of other adapter proteins including Shc, grb2, crk, crkl and nck and likely functions as a negative regulator of immunoreceptor signaling. We have demonstrated a number of specific molecules which are tyrosine phosphorylated following FcRI crosslinking, including the Shc adapter molecule and the Raf-1 protein kinase (J.Biol.Chem. 271:13342, 1996). Adapter proteins including Shc, grb2, crk, crkl, nck, etc. are involved in the translation of information


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by serving as a link between the cell surface receptor molecule (FcRI) and nucleotide exchange factors which activate the small GTPases, ras, rac ,and rap1a. Interestingly the respiratory burst is modulated by the conversion of GDPrac and GDPrap1a to GTPrac and GTPrap1a. We demonstrated a physical interaction between the FcRI subunit and the grb2 and cbl adapter proteins in U937IF cells (J.Immun, 161: 5555, 1998.). Many of these interactions are constitutive while others are induced by receptor activation. For example the Shc interacts with grb2 via the grb2-SH2 domain and only occurs upon FcRI receptor stimulation correlating with the tyrosine phosphorylation of Shc. In contrast, the grb2-cbl interaction is a constitutive one. Interestingly, we have demonstrated using GST-grb2 fusion constructs that there is a qualitative change in the grb2-cbl interaction following FcRI stimulation. In the resting cell, grb2 is bound to cbl via both the grb2-SH3-SH2 domains. Upon receptor activation, cbl is tyrosine phosphorylated and cbl is noted to bind to grb2 exclusively via the grb2-SH2 domain. We propose a proto-oncogenic role for the cbl molecule that suggests that cbl serves as an “adapter shield” regulating the capacity of the grb2 adapter protein to bind to the nucleotide exchange protein, SOS in the regulation of ras. We have prepared C-terminal and N-terminal fragments of the cbl molecule and have subcloned these constructs into the pGEX bacterial expression system in order to determine the interacting domains of cbl which mediate the cbl-grb2 interaction. So far our evidence suggests that this “adapter shield” function of cbl may extend to other adapter protein-cbl interaction e.g. cbl-crkl interaction. The general focus of these studies is to identify novel protein-protein interactions as targets for drug development for control of inflammation and immunoreceptor signaling. iii. Role of the small GTPases rac2 in FcRI and integrin signaling, A RAC2 knockout mouse model. The small GTPases are important components of a diverse array of signaling events in all cells. Our early experiments implicated RAS and RAC in FcR signaling in macrophages (J Immunol. 163: 6023, 1999). The rac family includes 3 genes, rac1, rac2 and rac3. Rac2 is specifically expressed in myeloid cells whereas the other two homologs are more ubiquitously expressed. We used a mouse genetics approach to determine the role of RAC2 in FcR and FcRI signaling in myeloid cells. We have examined signaling in the rac2 knockout mouse model and have characterized the homozygous -/- mice for FcR and FcR functions in macrophages and mast cells, respectively. The macrophages and mast cells from the homozygous knockout mice are deficient in Fc and FcR signaling. We use retroviral gene transfer to reconstitute Rac2 in the Rac2 deficient macrophages and mast cells to show that Rac2 fulfills a very specific function in macrophages and mast cells. We are currently focusing our efforts on determining the function of Rac2 in macrophage and mast cell signaling. As mentioned before the specificity of Rac2 phenotype suggests that rac2 and downstream pathways will be potentially useful drug targets for control of allergy, inflammation and angiogenesis (J. Biol. Chem, 2003). More recently, we have determined a role for Rac2 in endothelial cell function and angiogenesis in vivo (Exp Cell Res, 2008). We have discovered a new signaling axis, the v3-Syk-Rac2-Hif1-VEGF axis which is required for postnatal angiogenesis. This pathway has recently been reported by our group to regulate macrophage M1-M2 transition, tumor growth, metastasis and other M2 phenotypes (PLoS One, 2013). Drug discovery efforts built around Syk kinase screens in silico are ongoing. iv. Role of protein and lipid phosphatases in FcRI, ITIM and ITAM signaling. Our laboratory has had a long standing interest in phosphatase control of signal relay (Exp.Cell Res., 237: 288, 1997) Recent data from a number of laboratories have shown that ITAMs and ITIMs are critical for the proper regulation of immunoreceptor signaling. ITIMs or immunoreceptor tyrosine-based inhibitory motifs bind to phosphatases to shut off excitatory signals transmitted through ITAMs or immunoreceptor tyrosine-based activation motifs within immune cells i.e. macrophages, T cell, mast cell, platelets, etc. Controlling these signals can have dramatic effects upon the immune system allowing for immunomodulation of autoimmunity, graft rejection and inflammation. Recently we demonstrated that the SHP-1 phosphatase and SHIP regulate FcR signaling in macrophages. The molecular basis for this regulation appears to result from the control of Cbl phosphorylation status. In our studies these phosphatases function as a negative feedback loop to control Fc receptor signaling. It would be predicted that immune complex generated inflammation could be controlled by these signaling pathways. We reported that PTEN and SHP-1 controls the FcR


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induced phagocytosis and regulates Rac (Blood, 99: 694, 2002, Blood, 99: 1852, 2002)(Figure 2) again suggesting a potential important role for PI-3 kinase pathways in inflammation. More recently we implicated PKC as an activator of PTPase activity mechanistically linked to the downregulation of signaling through the FcR in macrophages (Exp Cell Res. 2013). v. Role of serine/threonine kinases in FcRI/ITIM/ITAM signaling. It is well established that upstream small GTPases act upon downstream serine/threonine kinase as effectors of signal relay. These include a large number of kinases each affecting a different set of downstream targets thus encoding a part of the specificity of signals transmitted through the Fc receptor. Importantly, the FcR generates a number of physiologic important signals from transcriptional activation of cytokine genes to mediation of the respiratory burst, phagocytosis, degranulation events, etc. Each response is encoded at the level of signal relay through specific downstream effectors. Certain effector kinases send activating signals whereas others engage negative feedback loops to shut off the receptor. We are interested in using this model to decipher the circuitry for positive and negative regulation of these signals. Importantly, these mechanisms are likely to be highly conserved hence an understanding of FcR signal regulation will provide general information regarding mechanisms of mammalian signal relay in particular related to the mechanisms of ITIM suppression of immunity and therapeutics around checkpoint inhibition (i.e. PD1 and BTK). We have now implicated Syk, Cbl, Shc, grb2, SOS, RAS, and PI-3 kinase in FcRI ST. (J.Biol.Chem. 271: 13342, 1996). In the FcRI system the Cbl protein is tyrosine phosphorylated following FcRI crosslinking to a higher stoichiometry than has been reported in other signaling systems. We have determined that PI-3 kinase is required for the assembly of the respiratory burst machinery and phagocytosis. The role of PI-3 kinase appears to be linked to the activation of RAC. Hence our current efforts are focused on the role of PI-3 kinase, PDK1/2 and AKT in regulation of RB. In addition, since RAC is known to regulate the PAK kinase this is another target signaling pathway for regulation of the respiratory burst (Exp. Cell Res. 245: 330, 1998). Below we provide a schematic diagram of some of the protein-protein interactions which our model predicts to be the basis for FcRI signaling to the respiratory burst.

Fig. 5. Model for ITAM and ITIM signaling in macrophages. As is now known the revolution in Oncology in 2014 came with the discovery that checkpoint blockade of ITIM receptors (CD28, PD1, PDL1) could activate the immune system for significant efficacy in the treatment of human cancer! Ligand (IgG) binds to the FcRI subunit resulting in a conformation change in the homodimeric ITAM or subunits. This change induces the activation of HCK kinase activity which results in the tyrosine phosphorylation of the ITAM motif of FcRI. Phosphorylation

of FcRI subunit recruits the binding and activation of the HCK, LYN and SYK kinases. Other proteins are tyrosine phosphorylated including the CBL and SHC adapter protein. The tyrosine phosphorylation of SHC is noted to bind to GRB2 (not shown) and the SOS nucleotide exchange protein, thus activating small GTPases in the cell through the conversion of GDPras to GTPras. GTPras activates downstream cascades including PI-3 kinase which generates PIP3 and activates other pathways. The role of CBL phosphorylation is to recruit to the receptor complex the PI-3 kinase p85 subunit. Downstream targets for FcRI stimulation are the small

RAS GTP

RAS GDP

PIP3

PDK-1

SOS

P-I 3 kinase

SYK

HCK/LYN

SHC

GRB2

FcRI

SH2SH2ITAM

IgGO2 O2

. -

gp91

p67p47

p22

rac

rap1aSH2

CBL

COOH

SH3

SH3

Respiratory BurstActivated Receptor complex

C

PIP3

p85

AKT

PTEN


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GTPases, RAS, RAP1A, and RAC2 which control the myeloid respiratory burst. The respiratory burst response involves the macromolecular assembly of p47phox, p67phox, p40phox, p91phox and p22phox along with RAP1A and RAC resulting in the generation of superoxide anions (O2-) which is measured using the reduction of cytochrome c as an assay. Abbreviations: , subunit (ITAM) of high affinity Fc receptor for IgG or FcRI subunit; SH2, src homology 2 domain; SH3, src homology 3 domain; SOS, "son of sevenless" protein. vi. New agents for immuno-oncology; Checkpoint blockade or TLR agonists to activate the immune response by blocking ITIM signaling in immune effectors (Fig 5). The Durden laboratory has been involved in the study of ITAM and ITIM signaling for the last 20 years. Finally, and perhaps most importantly our laboratory effort has recently identified the Rac2 and PI-3 kinase signaling pathways as negative regulators of the innate and adaptive immune response as potential control points for augmenting innate and adaptive immunity as an anticancer strategy. Moreover, we have executed the use of a TLR3 agonist, polyICLC in a Phase II trial in pediatric high grade glioma (funded by FDA RO1, Durden PI). This will be combined with checkpoint PD1 blockade in future clinical trials using PI-3 kinase inhibitors. https://www.clinicaltrials.gov/ct2/results?term=polyICLC+and+pediatrics&Search=Search III. Growth factor receptor signaling in endothelial and neuronal cells. During my post-doctoral training in the laboratory of Jonathan Cooper, I collaborated with Dr. Andrius Kaslauskas who was studying signal transduction through the receptor protein tyrosine kinase, PDGF receptor. We identified the two major autophosphorylation sites in the PDGF receptor subunit, Y751 (in the kinase insert domain) and Y857 (in the catalytic domain). The tyrosines were mutated to phenylalanine using site-directed mutagenesis and the mutant cDNAs were expressed in a dog epithelial cell line devoid of PDGF receptors. Introduction of the wild type PDGF receptor confers responsiveness to PDGF stimulation. We demonstrated that Y751 in the kinase insert region is important in the binding of PI-3 kinase and the activation of PI-3 kinase upon PDGF stimulation (Cell Regulation 2: 413, 1991). We showed that Y857 in the catalytic domain is required for the catalytic activity of the PDGF receptor and for the subsequent autophosphorylation of Y751 and binding of PI-3 kinase. The study of receptor PTKs continues to be an exciting area of investigation contributing greatly to our understanding of how protein tyrosine kinases are regulated in the cell. Our current focus is on neurotrophin receptor, TrkA signaling (discussed below). More recently we have performed experiments on 51 cosignaling through TrkA receptor. In the Rac2 knockout mice, mast cell TrkA/51 signaling is markedly deficient. Other experiments in our laboratory have defined a differential role for Src kinase and Cbl adapter protein phosphorylation in TrkA signaling (Exp Cell Res. 307: 1-14 2005). Src is required for neural differentiation whereas Cbl phosphorylation is associated with proliferation. The role of Src kinase in receptor tyrosine kinase signaling is an area of intense interest at this time. Again, our purpose is to decipher the TrkA signaling events involved in endothelial and neural proliferative responses versus differentiation and then develop drugs for therapeutic gain. Recently, we defined a role for the Csk and Src kinases in nuclear hormone receptor (RAR) signal relay in neuronal cells. It appears that Src family kinases are required for retinoic acid receptor signals required for differentiation (MCB, 27:4179, 2007). This may define a new paradigm for nuclear receptor signaling separate from the known genomic signaling pathways previously studied. These studies were funded via the NIH and relate to development of neuroblastoma therapeutics (CA81403). More recently we discovered that the transduction and overexpression of csk alone in astrocytes is sufficient to transform them into malignant glioma. The results support CSK kinase as a potent oncogene in this cell type. Drug discovery efforts are underway to isolate CSK inhibitors for therapeutic application.

Documents

CURRICULUM VITAE DONALD L. DURDEN, M.D., Ph.D.jacobsschool.ucsd.edu/uploads/agile_faculty/cv/durden_donald.pdfCurriculum vitae Donald L. Durden, M.D., Ph.D 04/28/16 6 2012-present