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