Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer

Yezi Zhu; Adam Sharp; Courtney M. Anderson; John L. Silberstein; Maritza Taylor; Changxue Lu; Pei Zhao; Angelo M. De Marzo; Emmanuel S. Antonarakis; Mindy Wang; Xingyong Wu; Yuling Luo; Nan Su; Daniel Nava Rodrigues; Ines Figueiredo; Jonathan Welti; Emily Park; Xiao Jun Ma; Ilsa Coleman; Colm Morrissey; Stephen R. Plymate; Peter S. Nelson; Johann S. de Bono; Jun Luo

doi:10.1016/j.eururo.2017.08.009

Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer

Yezi Zhu, Adam Sharp, Courtney M. Anderson, John L. Silberstein, Maritza Taylor, Changxue Lu, Pei Zhao, Angelo M. De Marzo, Emmanuel S. Antonarakis, Mindy Wang, Xingyong Wu, Yuling Luo, Nan Su, Daniel Nava Rodrigues, Ines Figueiredo, Jonathan Welti, Emily Park, Xiao Jun Ma, Ilsa Coleman, Colm MorrisseyStephen R. Plymate, Peter S. Nelson, Johann S. de Bono, Jun Luo

Research output: Contribution to journal › Article › peer-review

47 Scopus citations

Abstract

Background: Androgen receptor splice variant 7 (AR-V7) has been implicated in resistance to abiraterone and enzalutamide treatment in men with metastatic castration-resistant prostate cancer (mCRPC). Tissue- or cell-based in situ detection of AR-V7, however, has been limited by lack of specificity. Objective: To address current limitations in precision measurement of AR-V7 by developing a novel junction-specific AR-V7 RNA in situ hybridization (RISH) assay compatible with automated quantification. Design, setting, and participants: We designed a RISH method to visualize single splice junctions in cells and tissue. Using the validated assay for junction-specific detection of the full-length AR (AR-FL) and AR-V7, we generated quantitative data, blinded to clinical data, for 63 prostate tumor biopsies. Outcome measurements and statistical analysis: We evaluated clinical correlates of AR-FL/AR-V7 measurements, including association with prostate-specific antigen progression-free survival (PSA-PFS) and clinical and radiographic progression-free survival (PFS), in a subset of patients starting treatment with abiraterone or enzalutamide following biopsy. Results and limitations: Quantitative AR-FL/AR-V7 data were generated from 56 of the 63 (88.9%) biopsy specimens examined, of which 44 were mCRPC biopsies. Positive AR-V7 signals were detected in 34.1% (15/44) mCRPC specimens, all of which also co-expressed AR-FL. The median AR-V7/AR-FL ratio was 11.9% (range 2.7–30.3%). Positive detection of AR-V7 was correlated with indicators of high disease burden at baseline. Among the 25 CRPC biopsies collected before treatment with abiraterone or enzalutamide, positive AR-V7 detection, but not higher AR-FL, was significantly associated with shorter PSA-PFS (hazard ratio 2.789, 95% confidence interval 1.12–6.95; p = 0.0081). Conclusions: We report for the first time a RISH method for highly specific and quantifiable detection of splice junctions, allowing further characterization of AR-V7 and its clinical significance. Patient summary: Higher AR-V7 levels detected and quantified using a novel method were associated with poorer response to abiraterone or enzalutamide in prostate cancer. We developed and evaluated a novel method for splice junction–specific detection of the full-length AR (AR-FL) and splice variant AR-V7. Using this method, higher AR-V7, but not higher AR-FL, was associated with reduced clinical benefit to abiraterone or enzalutamide.

Original language	English (US)
Pages (from-to)	727-735
Number of pages	9
Journal	European Urology
Volume	73
Issue number	5
DOIs	https://doi.org/10.1016/j.eururo.2017.08.009
State	Published - May 2018
Externally published	Yes

Bibliographical note

Funding Information:
Funding/Support and role of the sponsor: Work by the Johns Hopkins University School of Medicine was supported by National Institutes of Health Grants R01 CA185297 and P30 CA006973, Department of Defense Prostate Cancer Research Program Grants W81XWH-15-2-0050, Johns Hopkins Prostate SPORE Grant P50 CA058236, and the Prostate Cancer Foundation. Work in the de Bono laboratory was supported by funding from the US Department of Defense, the Prostate Cancer Foundation, Movember, Prostate Cancer UK, Stand Up To Cancer, Cancer Research UK, and the UK Department of Health through an Experimental Cancer Medicine Centre grant. Work by the University of Washington and Fred Hutchinson Cancer Research Center was supported by the Department of Defense Prostate Cancer Research Program (W81XWH-14-2-0183, W81XWH-13-2-0093 and W81XWH-15-1-0430), Pacific Northwest Prostate Cancer SPORE (P50CA97186), a PO1 NIH grant (PO1CA163227), GRECC Veterans Affairs Research Service, the Institute for Prostate Cancer Research, and the Prostate Cancer Foundation. Adam Sharp is supported by the Medical Research Council, the Academy of Medical Sciences and Prostate Cancer UK. BaseScope assay-related materials were provided by Advanced Cell Diagnostics, Inc. (Newark, CA, USA). The sponsors played a role in manuscript approval.

Publisher Copyright:
© 2017 European Association of Urology

Keywords

AR-V7
Androgen receptor
RNA in situ hybridization
Splice variant

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.eururo.2017.08.009

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Zhu, Y., Sharp, A., Anderson, C. M., Silberstein, J. L., Taylor, M., Lu, C., Zhao, P., De Marzo, A. M., Antonarakis, E. S., Wang, M., Wu, X., Luo, Y., Su, N., Nava Rodrigues, D., Figueiredo, I., Welti, J., Park, E., Ma, X. J., Coleman, I., ... Luo, J. (2018). Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer. European Urology, 73(5), 727-735. https://doi.org/10.1016/j.eururo.2017.08.009

Zhu, Y, Sharp, A, Anderson, CM, Silberstein, JL, Taylor, M, Lu, C, Zhao, P, De Marzo, AM, Antonarakis, ES, Wang, M, Wu, X, Luo, Y, Su, N, Nava Rodrigues, D, Figueiredo, I, Welti, J, Park, E, Ma, XJ, Coleman, I, Morrissey, C, Plymate, SR, Nelson, PS, de Bono, JS & Luo, J 2018, 'Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer', European Urology, vol. 73, no. 5, pp. 727-735. https://doi.org/10.1016/j.eururo.2017.08.009

@article{ba32881aadd646d1ad1d9978197c8b40,

title = "Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer",

abstract = "Background: Androgen receptor splice variant 7 (AR-V7) has been implicated in resistance to abiraterone and enzalutamide treatment in men with metastatic castration-resistant prostate cancer (mCRPC). Tissue- or cell-based in situ detection of AR-V7, however, has been limited by lack of specificity. Objective: To address current limitations in precision measurement of AR-V7 by developing a novel junction-specific AR-V7 RNA in situ hybridization (RISH) assay compatible with automated quantification. Design, setting, and participants: We designed a RISH method to visualize single splice junctions in cells and tissue. Using the validated assay for junction-specific detection of the full-length AR (AR-FL) and AR-V7, we generated quantitative data, blinded to clinical data, for 63 prostate tumor biopsies. Outcome measurements and statistical analysis: We evaluated clinical correlates of AR-FL/AR-V7 measurements, including association with prostate-specific antigen progression-free survival (PSA-PFS) and clinical and radiographic progression-free survival (PFS), in a subset of patients starting treatment with abiraterone or enzalutamide following biopsy. Results and limitations: Quantitative AR-FL/AR-V7 data were generated from 56 of the 63 (88.9%) biopsy specimens examined, of which 44 were mCRPC biopsies. Positive AR-V7 signals were detected in 34.1% (15/44) mCRPC specimens, all of which also co-expressed AR-FL. The median AR-V7/AR-FL ratio was 11.9% (range 2.7–30.3%). Positive detection of AR-V7 was correlated with indicators of high disease burden at baseline. Among the 25 CRPC biopsies collected before treatment with abiraterone or enzalutamide, positive AR-V7 detection, but not higher AR-FL, was significantly associated with shorter PSA-PFS (hazard ratio 2.789, 95% confidence interval 1.12–6.95; p = 0.0081). Conclusions: We report for the first time a RISH method for highly specific and quantifiable detection of splice junctions, allowing further characterization of AR-V7 and its clinical significance. Patient summary: Higher AR-V7 levels detected and quantified using a novel method were associated with poorer response to abiraterone or enzalutamide in prostate cancer. We developed and evaluated a novel method for splice junction–specific detection of the full-length AR (AR-FL) and splice variant AR-V7. Using this method, higher AR-V7, but not higher AR-FL, was associated with reduced clinical benefit to abiraterone or enzalutamide.",

keywords = "AR-V7, Androgen receptor, RNA in situ hybridization, Splice variant",

author = "Yezi Zhu and Adam Sharp and Anderson, {Courtney M.} and Silberstein, {John L.} and Maritza Taylor and Changxue Lu and Pei Zhao and {De Marzo}, {Angelo M.} and Antonarakis, {Emmanuel S.} and Mindy Wang and Xingyong Wu and Yuling Luo and Nan Su and {Nava Rodrigues}, Daniel and Ines Figueiredo and Jonathan Welti and Emily Park and Ma, {Xiao Jun} and Ilsa Coleman and Colm Morrissey and Plymate, {Stephen R.} and Nelson, {Peter S.} and {de Bono}, {Johann S.} and Jun Luo",

note = "Funding Information: Funding/Support and role of the sponsor: Work by the Johns Hopkins University School of Medicine was supported by National Institutes of Health Grants R01 CA185297 and P30 CA006973, Department of Defense Prostate Cancer Research Program Grants W81XWH-15-2-0050, Johns Hopkins Prostate SPORE Grant P50 CA058236, and the Prostate Cancer Foundation. Work in the de Bono laboratory was supported by funding from the US Department of Defense, the Prostate Cancer Foundation, Movember, Prostate Cancer UK, Stand Up To Cancer, Cancer Research UK, and the UK Department of Health through an Experimental Cancer Medicine Centre grant. Work by the University of Washington and Fred Hutchinson Cancer Research Center was supported by the Department of Defense Prostate Cancer Research Program (W81XWH-14-2-0183, W81XWH-13-2-0093 and W81XWH-15-1-0430), Pacific Northwest Prostate Cancer SPORE (P50CA97186), a PO1 NIH grant (PO1CA163227), GRECC Veterans Affairs Research Service, the Institute for Prostate Cancer Research, and the Prostate Cancer Foundation. Adam Sharp is supported by the Medical Research Council, the Academy of Medical Sciences and Prostate Cancer UK. BaseScope assay-related materials were provided by Advanced Cell Diagnostics, Inc. (Newark, CA, USA). The sponsors played a role in manuscript approval. Publisher Copyright: {\textcopyright} 2017 European Association of Urology",

year = "2018",

month = may,

doi = "10.1016/j.eururo.2017.08.009",

language = "English (US)",

volume = "73",

pages = "727--735",

journal = "European Urology",

issn = "0302-2838",

publisher = "Elsevier",

number = "5",

}

TY - JOUR

T1 - Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer

AU - Zhu, Yezi

AU - Sharp, Adam

AU - Anderson, Courtney M.

AU - Silberstein, John L.

AU - Taylor, Maritza

AU - Lu, Changxue

AU - Zhao, Pei

AU - De Marzo, Angelo M.

AU - Antonarakis, Emmanuel S.

AU - Wang, Mindy

AU - Wu, Xingyong

AU - Luo, Yuling

AU - Su, Nan

AU - Nava Rodrigues, Daniel

AU - Figueiredo, Ines

AU - Welti, Jonathan

AU - Park, Emily

AU - Ma, Xiao Jun

AU - Coleman, Ilsa

AU - Morrissey, Colm

AU - Plymate, Stephen R.

AU - Nelson, Peter S.

AU - de Bono, Johann S.

AU - Luo, Jun

N1 - Funding Information: Funding/Support and role of the sponsor: Work by the Johns Hopkins University School of Medicine was supported by National Institutes of Health Grants R01 CA185297 and P30 CA006973, Department of Defense Prostate Cancer Research Program Grants W81XWH-15-2-0050, Johns Hopkins Prostate SPORE Grant P50 CA058236, and the Prostate Cancer Foundation. Work in the de Bono laboratory was supported by funding from the US Department of Defense, the Prostate Cancer Foundation, Movember, Prostate Cancer UK, Stand Up To Cancer, Cancer Research UK, and the UK Department of Health through an Experimental Cancer Medicine Centre grant. Work by the University of Washington and Fred Hutchinson Cancer Research Center was supported by the Department of Defense Prostate Cancer Research Program (W81XWH-14-2-0183, W81XWH-13-2-0093 and W81XWH-15-1-0430), Pacific Northwest Prostate Cancer SPORE (P50CA97186), a PO1 NIH grant (PO1CA163227), GRECC Veterans Affairs Research Service, the Institute for Prostate Cancer Research, and the Prostate Cancer Foundation. Adam Sharp is supported by the Medical Research Council, the Academy of Medical Sciences and Prostate Cancer UK. BaseScope assay-related materials were provided by Advanced Cell Diagnostics, Inc. (Newark, CA, USA). The sponsors played a role in manuscript approval. Publisher Copyright: © 2017 European Association of Urology

PY - 2018/5

Y1 - 2018/5

N2 - Background: Androgen receptor splice variant 7 (AR-V7) has been implicated in resistance to abiraterone and enzalutamide treatment in men with metastatic castration-resistant prostate cancer (mCRPC). Tissue- or cell-based in situ detection of AR-V7, however, has been limited by lack of specificity. Objective: To address current limitations in precision measurement of AR-V7 by developing a novel junction-specific AR-V7 RNA in situ hybridization (RISH) assay compatible with automated quantification. Design, setting, and participants: We designed a RISH method to visualize single splice junctions in cells and tissue. Using the validated assay for junction-specific detection of the full-length AR (AR-FL) and AR-V7, we generated quantitative data, blinded to clinical data, for 63 prostate tumor biopsies. Outcome measurements and statistical analysis: We evaluated clinical correlates of AR-FL/AR-V7 measurements, including association with prostate-specific antigen progression-free survival (PSA-PFS) and clinical and radiographic progression-free survival (PFS), in a subset of patients starting treatment with abiraterone or enzalutamide following biopsy. Results and limitations: Quantitative AR-FL/AR-V7 data were generated from 56 of the 63 (88.9%) biopsy specimens examined, of which 44 were mCRPC biopsies. Positive AR-V7 signals were detected in 34.1% (15/44) mCRPC specimens, all of which also co-expressed AR-FL. The median AR-V7/AR-FL ratio was 11.9% (range 2.7–30.3%). Positive detection of AR-V7 was correlated with indicators of high disease burden at baseline. Among the 25 CRPC biopsies collected before treatment with abiraterone or enzalutamide, positive AR-V7 detection, but not higher AR-FL, was significantly associated with shorter PSA-PFS (hazard ratio 2.789, 95% confidence interval 1.12–6.95; p = 0.0081). Conclusions: We report for the first time a RISH method for highly specific and quantifiable detection of splice junctions, allowing further characterization of AR-V7 and its clinical significance. Patient summary: Higher AR-V7 levels detected and quantified using a novel method were associated with poorer response to abiraterone or enzalutamide in prostate cancer. We developed and evaluated a novel method for splice junction–specific detection of the full-length AR (AR-FL) and splice variant AR-V7. Using this method, higher AR-V7, but not higher AR-FL, was associated with reduced clinical benefit to abiraterone or enzalutamide.

AB - Background: Androgen receptor splice variant 7 (AR-V7) has been implicated in resistance to abiraterone and enzalutamide treatment in men with metastatic castration-resistant prostate cancer (mCRPC). Tissue- or cell-based in situ detection of AR-V7, however, has been limited by lack of specificity. Objective: To address current limitations in precision measurement of AR-V7 by developing a novel junction-specific AR-V7 RNA in situ hybridization (RISH) assay compatible with automated quantification. Design, setting, and participants: We designed a RISH method to visualize single splice junctions in cells and tissue. Using the validated assay for junction-specific detection of the full-length AR (AR-FL) and AR-V7, we generated quantitative data, blinded to clinical data, for 63 prostate tumor biopsies. Outcome measurements and statistical analysis: We evaluated clinical correlates of AR-FL/AR-V7 measurements, including association with prostate-specific antigen progression-free survival (PSA-PFS) and clinical and radiographic progression-free survival (PFS), in a subset of patients starting treatment with abiraterone or enzalutamide following biopsy. Results and limitations: Quantitative AR-FL/AR-V7 data were generated from 56 of the 63 (88.9%) biopsy specimens examined, of which 44 were mCRPC biopsies. Positive AR-V7 signals were detected in 34.1% (15/44) mCRPC specimens, all of which also co-expressed AR-FL. The median AR-V7/AR-FL ratio was 11.9% (range 2.7–30.3%). Positive detection of AR-V7 was correlated with indicators of high disease burden at baseline. Among the 25 CRPC biopsies collected before treatment with abiraterone or enzalutamide, positive AR-V7 detection, but not higher AR-FL, was significantly associated with shorter PSA-PFS (hazard ratio 2.789, 95% confidence interval 1.12–6.95; p = 0.0081). Conclusions: We report for the first time a RISH method for highly specific and quantifiable detection of splice junctions, allowing further characterization of AR-V7 and its clinical significance. Patient summary: Higher AR-V7 levels detected and quantified using a novel method were associated with poorer response to abiraterone or enzalutamide in prostate cancer. We developed and evaluated a novel method for splice junction–specific detection of the full-length AR (AR-FL) and splice variant AR-V7. Using this method, higher AR-V7, but not higher AR-FL, was associated with reduced clinical benefit to abiraterone or enzalutamide.

KW - AR-V7

KW - Androgen receptor

KW - RNA in situ hybridization

KW - Splice variant

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JO - European Urology

JF - European Urology

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ER -

Novel Junction-specific and Quantifiable In Situ Detection of AR-V7 and its Clinical Correlates in Metastatic Castration-resistant Prostate Cancer

Abstract

Bibliographical note

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UN SDGs

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