Role of androgen receptor splice variant-7 (AR-V7) in prostate cancer resistance to 2nd-generation androgen receptor signaling inhibitors


The role of truncated androgen receptor splice variant-7 (AR-V7) in prostate cancer biology is an unresolved question. Is it simply a marker of resistance to 2nd-generation androgen receptor signaling inhibitors (ARSi) like abiraterone acetate (Abi) and enzalutamide (Enza) or a functional driver of lethal resistance via its ligand-independent transcriptional activity? To resolve this question, the correlation between resistance to ARSi and genetic chances and expression of full length AR (AR-FL) vs. AR-V7 were evaluated in a series of independent patient-derived xenografts (PDXs). While all PDXs lack PTEN expression, there is no consistent requirement for mutation in TP53, RB1, BRCA2, PIK3CA, or MSH2, or expression of SOX2 or ERG and ARSi resistance. Elevated expression of AR-FL alone is sufficient for Abi but not Enza resistance, even if AR-FL is gain-of-function (GOF) mutated. Enza resistance is consistently correlated with enhanced AR-V7 expression. In vitro and in vivo growth responses of Abi-/Enza-resistant LNCaP-95 cells in which CRISPR-Cas9 was used to knockout AR-FL or AR-V7 alone or in combination were evaluated. Combining these growth responses with RNAseq analysis demonstrates that both AR-FL- and AR-V7-dependent transcriptional complementation are needed for Abi/Enza resistance.

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Fig. 1: Characterization of CWR22-RH.
Fig. 2: RNA-seq-based expression analysis of a subset of genes across PDX models.
Fig. 3: Characterization of LvCaP-2 and LvCaP-2R.
Fig. 4: Characterization of SkCaP-1 and SkCaP-1R.
Fig. 5: Characterization of LNCaP variant under long-term ARSi-equivalent conditions (i.e., LN-95 cells).
Fig. 6: Characterization of AR-FL, AR-V7, vs. total AR knockout in LN-95 cells in vitro.
Fig. 7: Characterization of AR-FL, AR-V7, vs. total AR knockout in LN-95 cells in vivo.


  1. 1.

    Kurita T, Wang YZ, Donjacour AA, Zhao C, Lydon JP, O’Malley BW, et al. Paracrine regulation of apoptosis by steroid hormones in the male and female reproductive system. Cell Death Differ. 2001;8:192–200.

    CAS  Google Scholar 

  2. 2.

    Litvinov IV, De Marzo AM, Isaacs JT. Is the Achilles’ heel for prostate cancer therapy a gain of function in androgen receptor signaling? J Clin Endocrinol Metab. 2003;88:2972–82.

    CAS  Google Scholar 

  3. 3.

    Isaacs JT. Resolving the Coffey Paradox: what does the androgen receptor do in normal vs. malignant prostate epithelial cells? Am J Clin Exp Urol. 2018;6:55–61.

    Google Scholar 

  4. 4.

    Vander Griend DJ, D’Antonio J, Gurel B, Antony L, Demarzo AM, Isaacs JT. Cell-autonomous intracellular androgen receptor signaling drives the growth of human prostate cancer initiating cells. Prostate. 2010;70:90–99.

    Google Scholar 

  5. 5.

    Kyprianou N, English HF, Isaacs JT. Programmed cell death during regression of PC-82 human prostate cancer following androgen ablation. Cancer Res. 1990;50:3748–53.

    CAS  Google Scholar 

  6. 6.

    Sharp A, Welti J, Blagg J, de Bono JS. Targeting androgen receptor aberrations in castration-resistant prostate cancer. Clin Cancer Res. 2016;22:4280–2.

    CAS  Google Scholar 

  7. 7.

    Antonarakis ES, Lu C, Wang H, Luber B, Nakazawa M, Roeser JC, et al. AR-V7 and resistance to enzalutamide and abiraterone in prostate cancer. N Engl J Med. 2014;371:1028–38.

    Google Scholar 

  8. 8.

    Dehm SM, Schmidt LJ, Heemers HV, Vessella RL, Tindall DJ. Splicing of a novel androgen receptor exon generates a constitutively active androgen receptor that mediates prostate cancer therapy resistance. Cancer Res. 2008;68:5469–77.

    CAS  Google Scholar 

  9. 9.

    Guo Z, Yang X, Sun F, Jiang R, Linn DE, Chen H, et al. A novel androgen receptor splice variant is up-regulated during prostate cancer progression and promotes androgen depletion-resistant growth. Cancer Res. 2009;69:2305–13.

    CAS  Google Scholar 

  10. 10.

    Hu R, Dunn TA, Wei S, Isharwal S, Veltri RW, Humphreys E, et al. Ligand-independent androgen receptor variants derived from splicing of cryptic exons signify hormone-refractory prostate cancer. Cancer Res. 2009;69:16–22.

    CAS  Google Scholar 

  11. 11.

    Lu C, Brown LC, Antonarakis ES, Armstrong AJ, Luo J. Androgen receptor variant-driven prostate cancer II: advances in laboratory investigations. Prostate Cancer Prostatic Dis. 2020;23:381–97.

    Article  Google Scholar 

  12. 12.

    Tan J, Sharief Y, Hamil KG, Gregory CW, Zang DY, Sar M, et al. Dehydroepiandrosterone activates mutant androgen receptors expressed in the androgen-dependent human prostate cancer xenograft CWR22 and LNCaP cells. Mol Endocrinol. 1997;11:450–9.

    CAS  Google Scholar 

  13. 13.

    Isaacs JT, D’Antonio JM, Chen S, Antony L, Dalrymple SP, Ndikuyeze GH, et al. Adaptive auto-regulation of androgen receptor provides a paradigm shifting rationale for bipolar androgen therapy (BAT) for castrate resistant human prostate cancer. Prostate. 2012;72:1491–505.

    CAS  Google Scholar 

  14. 14.

    Chen CD, Welsbie DS, Tran C, Baek SH, Chen R, Vessella R, et al. Molecular determinants of resistance to antiandrogen therapy. Nat Med. 2004;10:33–9.

    Google Scholar 

  15. 15.

    Chen EJ, Sowalsky AG, Gao S, Cai C, Voznesensky O, Schaefer R, et al. Abiraterone treatment in castration-resistant prostate cancer selects for progesterone responsive mutant androgen receptors. Clin Cancer Res. 2015;21:1273–80.

    CAS  Google Scholar 

  16. 16.

    Mostaghel EA, Zhang A, Hernandez S, Marck BT, Zhang X, Tamae D, et al. Contribution of adrenal glands to intratumor androgens and growth of castration-resistant prostate cancer. Clin Cancer Res. 2019;25:426–39.

    CAS  Google Scholar 

  17. 17.

    Lam HM, McMullin R, Nguyen HM, Coleman I, Gormley M, Gulati R, et al. Characterization of an abiraterone ultraresponsive phenotype in castration-resistant prostate cancer patient-derived xenografts. Clin Cancer Res. 2017;23:2301–12.

    CAS  Google Scholar 

  18. 18.

    Tran C, Ouk S, Clegg NJ, Chen Y, Watson PA, Arora V, et al. Development of a second-generation antiandrogen for treatment of advanced prostate cancer. Science. 2009;324:787–90.

    CAS  Google Scholar 

  19. 19.

    Moll JM, Kumagai J, van Royen ME, Teubel WJ, van Soest RJ, French PJ, et al. A bypass mechanism of abiraterone-resistant prostate cancer: Accumulating CYP17A1 substrates activate androgen receptor signaling. Prostate. 2019;79:937–48.

    CAS  Google Scholar 

  20. 20.

    Bellur S, Van der Kwast T, Mete O. Evolving concepts in prostatic neuroendocrine manifestations: from focal divergent differentiation to amphicrine carcinoma. Hum Pathol. 2018;85:313–27.

    Google Scholar 

  21. 21.

    Naidu CK, Suneetha Y. Prediction and analysis of breast cancer related deleterious non-synonymous single nucleotide polymorphisms in the PTEN gene. Asian Pac J Cancer Prev. 2016;17:2199–203.

    Google Scholar 

  22. 22.

    Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H, Chu TM, et al. LNCaP model of human prostatic carcinoma. Cancer Res. 1983;43:1809–18.

    CAS  Google Scholar 

  23. 23.

    Li J, Yen C, Liaw D, Podsypanina K, Bose S, Wang SI, et al. PTEN, a putative protein tyrosine phosphatase gene mutated in human brain. Breast, Prostate Cancer Sci. 1997;275:1943–7.

    CAS  Google Scholar 

  24. 24.

    Veldscholte J, Ris-Stalpers C, Kuiper GG, Jenster G, Berrevoets C, Claassen E, et al. A mutation in the ligand binding domain of the androgen receptor of human LNCaP cells affects steroid binding characteristics and response to anti-androgens. Biochem Biophys Res Commun. 1990;173:534–40.

    CAS  Google Scholar 

  25. 25.

    Guo Y, Kyprianou N. Restoration of transforming growth factor beta signaling pathway in human prostate cancer cells suppresses tumorigenicity via induction of caspase-1-mediated apoptosis. Cancer Res. 1999;59:1366–71.

    CAS  Google Scholar 

  26. 26.

    Lin X, Tascilar M, Lee WH, Vles WJ, Lee BH, Veeraswamy R, et al. GSTP1 CpG island hypermethylation is responsible for the absence of GSTP1 expression in human prostate cancer cells. Am J Pathol. 2001;159:1815–26.

    CAS  Google Scholar 

  27. 27.

    Zhang Q, Rubenstein JN, Jang TL, Pins M, Javonovic B, Yang X, et al. Insensitivity to transforming growth factor-beta results from promoter methylation of cognate receptors in human prostate cancer cells (LNCaP). Mol Endocrinol. 2005;19:2390–9.

    CAS  Google Scholar 

  28. 28.

    Berchem GJ, Bosseler M, Sugars LY, Voeller HJ, Zeitlin S, Gelmann EP. Androgens induce resistance to bcl-2-mediated apoptosis in LNCaP prostate cancer cells. Cancer Res. 1995;55:735–8.

    CAS  Google Scholar 

  29. 29.

    Esquenet M, Swinnen JV, Heyns W, Verhoeven G. LNCaP prostatic adenocarcinoma cells derived from low and high passage numbers display divergent responses not only to androgens but also to retinoids. J Steroid Biochem Mol Biol. 1997;62:391–9.

    CAS  Google Scholar 

  30. 30.

    Tombal B, Denmeade SR, Gillis JM, Isaacs JT. A supramicromolar elevation of intracellular free calcium ([Ca(2+)](i)) is consistently required to induce the execution phase of apoptosis. Cell Death Differ. 2002;9:561–73.

    CAS  Google Scholar 

  31. 31.

    Sedelaar JP, Isaacs JT. Tissue culture media supplemented with 10% fetal calf serum contains a castrate level of testosterone. Prostate. 2009;69:1724–9.

    CAS  Google Scholar 

  32. 32.

    Leach FS, Velasco A, Hsieh JT, Sagalowsky AI, McConnell JD. The mismatch repair gene hMSH2 is mutated in the prostate cancer cell line LNCaP. J Urol. 2000;164:1830–3.

    CAS  Google Scholar 

  33. 33.

    Karan D, Schmied BM, Dave BJ, Wittel UA, Lin MF, Batra SK. Decreased androgen-responsive growth of human prostate cancer is associated with increased genetic alterations. Clin Cancer Res. 2001;7:3472–80.

    CAS  Google Scholar 

  34. 34.

    Pflug BR, Reiter RE, Nelson JB. Caveolin expression is decreased following androgen deprivation in human prostate cancer cell lines. Prostate. 1999;40:269–73.

    CAS  Google Scholar 

  35. 35.

    Hu R, Lu C, Mostaghel EA, Yegnasubramanian S, Gurel M, Tannahill C, et al. Distinct transcriptional programs mediated by the ligand-dependent full-length androgen receptor and its splice variants in castration-resistant prostate cancer. Cancer Res. 2012;72:3457–62.

    CAS  Google Scholar 

  36. 36.

    Yang YC, Banuelos CA, Mawji NR, Wang J, Kato M, Haile S, et al. Targeting androgen receptor activation function-1 with EPI to overcome resistance mechanisms in castration-resistant prostate cancer. Clin Cancer Res. 2016;22:4466–77.

    CAS  Google Scholar 

  37. 37.

    Mostaghel EA, Cho E, Zhang A, Alyamani M, Kaipainen A, Green S, et al. Association of tissue abiraterone levels and SLCO genotype with intraprostatic steroids and pathologic response in men with high-risk localized prostate cancer. Clin Cancer Res. 2017;23:4592–601.

    CAS  Google Scholar 

  38. 38.

    Jenster G, Trapman J, Brinkmann AO. Nuclear import of the human androgen receptor. Biochem J. 1993;293:761–8.

    CAS  Google Scholar 

  39. 39.

    Liu LL, Xie N, Sun S, Plymate S, Mostaghel E, Dong X. Mechanisms of the androgen receptor splicing in prostate cancer cells. Oncogene. 2014;33:3140–50.

    CAS  Google Scholar 

  40. 40.

    Li Y, Chan SC, Brand LJ, Hwang TH, Silverstein KA, Dehm SM. Androgen receptor splice variants mediate enzalutamide resistance in castration-resistant prostate cancer cell lines. Cancer Res. 2013;73:483–9.

    CAS  Google Scholar 

  41. 41.

    Visakorpi T, Hyytinen E, Koivisto P, Tanner M, Keinanen R, Palmberg C, et al. In vivo amplification of the androgen receptor gene and progression of human prostate cancer. Nat Genet. 1995;9:401–6.

    CAS  Google Scholar 

  42. 42.

    Quigley DA, Dang HX, Zhao SG, Lloyd P, Aggarwal R, Alumkal JJ, et al. Genomic hallmarks and structural variation in metastatic prostate. Cancer Cell. 2018;175:889.

    CAS  Google Scholar 

  43. 43.

    Takeda DY, Spisak S, Seo JH, Bell C, O’Connor E, Korthauer K, et al. A somatically acquired enhancer of the androgen receptor is a noncoding driver in advanced prostate. Cancer Cell. 2018;174:422–32. e413.

    CAS  Google Scholar 

  44. 44.

    Viswanathan SR, Ha G, Hoff AM, Wala JA, Carrot-Zhang J, Whelan CW, et al. Structural alterations driving castration-resistant prostate cancer revealed by linked-read genome sequencing. Cell. 2018;174:433–47. e19.

    CAS  Google Scholar 

  45. 45.

    Ajiboye AS, Esopi D, Yegnasubramanian S, Denmeade SR. Androgen receptor splice variants are not substrates of nonsense-mediated decay. Prostate. 2017;77:829–37.

    CAS  Google Scholar 

  46. 46.

    Zhu Y, Luo J. Regulation of androgen receptor variants in prostate cancer. Asian J Urol. 2020; in press.

  47. 47.

    Teply BA, Wang H, Luber B, Sullivan R, Rifkind I, Bruns A, et al. Bipolar androgen therapy in men with metastatic castration-resistant prostate cancer after progression on enzalutamide: an open-label, phase 2, multicohort study. Lancet Oncol. 2018;19:76–86.

    CAS  Google Scholar 

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We are grateful to the patients and their families who participated in the Legacy Gift Rapid Autopsy program at Hopkins. We would like to acknowledge the Department of Defense Prostate Cancer Research Program W81XWH1810349 (JTI), W81XWH-17-1-0528 (WNB), W81XWH-18-2-0015 (AMM) and NIH Prostate SPOREs P50 CA058236 (JTI), Pathology Core from the Prostate SPORE P50 CA058236 (AMM) and P50 CA097186 (PSN), W81XWH-18-1-0347 (PSN), P01 CA163227 (PSN), Emerson Collective Cancer Research Fund [643396, (WNB)], Allegheny Health Network-Johns Hopkins University Cancer Research Fund (WNB), R01 CA185297 (JL and ESA). Also, we wish to thank the Cell Imaging Facility, Animal Core Facility, Tissue Histology Core, Genetic Resource Core, Cytogenetics Core Facility, and the Autopsy Core from the CCSG Grant supported by the SKCCC CCSG (P30 CA006973) for their services and assistance.

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Correspondence to John T. Isaacs.

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ESA is a paid consultant/advisor to Janssen, Astellas, Sanofi, Dendreon, Pfizer, Amgen, AstraZeneca, Bristol Myers Squibb, Bayer, Clovis, and Merck; has received research funding (to his institution) from Janssen, Johnson & Johnson, Sanofi, Dendreon, Genentech, Novartis, Bristol Myers Squibb, AstraZeneca, Clovis, and Merck. ESA and JL are co-inventors of an AR-V7 biomarker technology that has been licensed to Qiagen.

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Zhu, Y., Dalrymple, S.L., Coleman, I. et al. Role of androgen receptor splice variant-7 (AR-V7) in prostate cancer resistance to 2nd-generation androgen receptor signaling inhibitors. Oncogene 39, 6935–6949 (2020).

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