Identification of the New Identification of the New Human Retrovirus XMRVHuman Retrovirus XMRVIdentification of the New Identification of the New
Human Retrovirus XMRVHuman Retrovirus XMRV
Disclosures
• Research Support– Abbott Laboratories
• Patent Licenses and Consulting– Abbott Laboratories
The Hereditary Prostate Cancer 1 Gene The Hereditary Prostate Cancer 1 Gene Encodes the Antiviral Protein RNase LEncodes the Antiviral Protein RNase L
Carpten, J.. et al., Nature Genetics, 30, 181-4, 2002
Prostate CancerTissue
RNA/DNAVirochip
Hybridization Pattern & RT- PCR
Urisman, et al. PLoS Pathogens 2006
Red VP= homozygous for R462Q RNase L Variant
Retrovirus Discovery in Prostate CancerRetrovirus Discovery in Prostate Cancer
Related toMurine LeukemiaVirus
Silverman et al., NATURE REVIEWS Urology, 2010
Urisman et al., PLoS Pathogen, 2006;Dong et al., PNAS, 2007
XMRVXMRVXenotropic Murine Leukemia Virus-Related Virus
Green: XMRVBlue: DAPIRed: Cytokeratin
Scale Bar=10m
XMRV in Prostate Cancer Tissue
Urisman A. PLoS Pathog. 2006 Mar;2(3):e25
Stromal fibroblast
Dividing stromal cell
Stromal hematopoetic cell
Semen-Derived Factor (SEVI) Enhances Infectivity
Hong, Klein et al, J Virol 83:6995, 2009
XMRV in human prostatic secretionssemen enhances infections
Effect of Androgen on XMRV
Androgen Stimulates XMRV Replication Casodex Suppresses XMRV Replication
ARE in XMRV Promoter
Dong & Silverman, J Virol 84:1648, 2010
•PCR and sequencing novel XMRV isolates
•XMRV proteins detected in activated PBMC, T cells and B cells (immunoblots)
•Activated PBMC from CFS patients, but not from controls, transmitted XMRV to LNCaP cells in co-culture experiments
•Plasma from CFS patients, but not from controls, transmitted XMRV to LNCaP cells
•Circulating antibody against XMRV envelope protein was detected in CFS patients, but not in control individuals
XMRV in CFS Cases [Lombardi et al., Science (2009)]
XMRV in Chronic Fatigue Syndrome
• XMRV detected by nPCR in 67% of patients vs. 3.7% of controls (n = 101 and 218)
Lombardi et al, Science 326:585, 2009
Electron Micrograph of C-type Retrovirus Particles Transmitted from CFS activated T cells to LNCaP
Association of XMRV with prostate cancer and other diseases
XMRV RNase L Methods Cell Types or fluidProstate Cancer R462QUrisman et al. (2006) Yes Yes Virochip,RT-PCR, FISH, IHC StromaDong et al. (2007) Yes Yes Integration site mapping n.d.Kim et al. (2008) Yes Yes Integration site mapping n.d.Fischer et al. (2008) Yes (low) No RT-PCR n.d.Sfanos et al. (2008) No n.d. PCR n.d.Knouf et al. (2009) Yes Yes RT-PCR epithelial (22Rv1)Hong et al. (2009) Yes No qRT-PCR, nested RT-PCR Prostate secretionsSchalberg et al. (2009) Yes No qPCR, IHC EpithelialHohn et al. (2009) No No PCR, qRT-PCR, ELISA n.d.Arnold et al. (2010) Yes Yes ab, IHC, FISH Stromal fibroblastsD’Arcy (2008) No No RT-PCR n.d.Furuta (2009) Yes n.d. Immune response, RT-PCR PBMC
SUMMARY: 9/12 5/10
Chronic fatigue syndromeLombardi et al. Yes No PCR, WB, culturing virus, ab PBMC, T-, B-cells, plasmaErlwein et al. No n.d. PCR whole bloodGroom et al. No n.d. PCR, ab PBMC, seraVan Kuppeveld et al. No n.d. PCR PBMCSwitzer et al. No n.d. PCR, WB, ELISA PBMC, sera
SUMMARY: 1/5 0/1
Amyotrophic lateral sclerosisMcCormick et al. No No RT-PCR PBMC, CSF
Large Differences in Detection RatesLarge Differences in Detection Rates(possible reasons)(possible reasons)
•Lab contaminationLab contamination
•Geographical distributionGeographical distribution
•Sequence variantsSequence variants
•Clinical criteria for patient selectionClinical criteria for patient selection
•Lack of standardized methodsLack of standardized methods
•Lack of widely available positive Lack of widely available positive control human specimenscontrol human specimens
Animal Protocol:
IV Inoculation of Rhesus Macaques with XMRV
-2 M -1 days 0 7 14 21 28 35 42 56 70 ....144 160 174 188 202 216 275 291
XMRV i.v.
Baseline Bleeds
Blood collections
XMRV vaccine
Necropsies: xx x xxRLm-1 RLq-10 RIl-10
ROu-4 RYh-10
XMRV reinfection
i.v. n=2
LN coll. x x x x x
XMRV Infection in PrimatesEarly Targets Late Targets
Lymphoid Organs
Peripheral Blood (CD4+)
Spleen & Lymph Nodes
Bone Marrow
Large & Small Intestine
Lymphoid Organs
---
Spleen & Lymph Nodes
---
Large & Small Intestine
Lung (macrophages) Lung (macrophages)
Pancreas (epithelium) ---
Genital Tract
Prostate (epithelium)
Testis
Seminal vesicle
Genital Tract
Prostate (stroma only)
Testis
Seminal vesicle & Epididymis
Cervix & Vagina (epithelium)
Possible Role of XMRV in Prostate Cancer and CFS
Silverman et al., NATURE REVIEWS Urology, 2010
Conclusions
• XMRV is a novel retrovirus associated with prostate cancer and CFS in
humans in some, but not all, studies
– All individuals at risk regardless of RNaseL/HPC1 genotype
• XMRV establishes both acute and chronic infections in primates
– Prostate epithelium is an early target; stroma late
– CD4+ and other blood cell types infected
• XMRV growth is fueled by androgen
_ Potential oncogenic mechanism
• XMRV might be transmitted by blood transfusion
– CFS pts deferred from donation in Canada, Australia, and NZ
• Causal link to any human disease remains to be established
AcknowledgementsAcknowledgements
Cleveland Clinic–Eric Klein–Jaydip Das Gupta–Beihua Dong–Ross Molinaro (Emory)
Abbott Diagnostics-John Hackett Jr-Gerald Schochetman-Xiaoxing Qui-Sushil Devare
Yerkes/Emory–Francois Villinger–Prachi Sharma
TGEN -John Carpten, Jeff TrentJHMI -William IsaacsUSC - Graham Casey
Whittemore Peterson Institute- Judy Mikovits, Vincent Lombardi
NCI- Francis Ruscetti, Sandra Ruscetti
University of Ulm - Jan Munch, Frank Kirchhoff
UCSF–Joseph DeRisi–Don Ganem–Anatoly Urisman–Nicole Fischer