Constitutively-active androgen receptor variants function independently of the HSP90 chaperone but do not confer resistance to HSP90 inhibitors

Joanna L Gillis,Wayne D Tilley,Luke A Selth,Stephen R Plymate,Scott L Townley,Margaret M Centenera,Shihua Sun,Lisa M Butler

doi:10.18632/oncotarget.975

Abstract

The development of lethal, castration resistant prostate cancer is associated with adaptive changes to the androgen receptor (AR), including the emergence of mutant receptors and truncated, constitutively active AR variants. AR relies on the molecular chaperone HSP90 for its function in both normal and malignant prostate cells, but the requirement for HSP90 in environments with aberrant AR expression is largely unknown. Here, we investigated the efficacy of three HSP90 inhibitors, 17-AAG, HSP990 and AUY922, against clinically-relevant AR missense mutants and truncated variants. HSP90 inhibition effectively suppressed the signaling of wild-type AR and all AR missense mutants tested. By contrast, two truncated AR variants, AR-V7 and ARv567es, exhibited marked resistance to HSP90 inhibitors. Supporting this observation, nuclear localization of the truncated AR variants was not affected by HSP90 inhibition and AR variant:HSP90 complexes could not be detected in prostate cancer cells. Interestingly, HSP90 inhibition resulted in accumulation of AR-V7 and ARv567es in both cell lines and human tumor explants. Despite the apparent independence of AR variants from HSP90 and their treatment-associated induction, the growth of cell lines with endogenous or enforced expression of AR-V7 or ARv567es remained highly sensitive to AUY922. This study demonstrates that functional AR variant signaling does not confer resistance to HSP90 inhibition, yields insight into the interaction between AR and HSP90 and provides further impetus for the clinical application of HSP90 inhibitors in advanced prostate cancer.

Highlights

castration-resistant prostate cancer (CRPC) is characterized by sustained androgen receptor (AR) signaling within the “castrate” environment [1]through adaptive mechanisms such as increased intratumoral androgen biosynthesis [2], AR overexpression and amplification [3, 4] and increased response to or levels of AR co-regulators [5]
We examined the efficacy of Heat shock protein 90 (HSP90) inhibitors against a diverse range of androgen receptor mutants and variants
HSP90 inhibitors repress the transcriptional activity of wild-type AR and a wide range of clinically-relevant AR mutants

Summary

Introduction

CRPC is characterized by sustained androgen receptor (AR) signaling within the “castrate” environment [1]through adaptive mechanisms such as increased intratumoral androgen biosynthesis [2], AR overexpression and amplification [3, 4] and increased response to or levels of AR co-regulators [5]. The isolation of constitutively active, truncated forms of the AR has revealed another mechanism underlying persistent AR signaling in CRPC. These socalled AR variants (ARVs), which arise due to aberrant splicing and/or structural rearrangements of the AR gene [11, 12], have variable structures but each lacks all or a portion of the ligand-binding domain (LBD) [13]. Two of the most commonly occurring variants, ARv567es and AR-V7, are induced by castration and their expression in bone metastases of men with CRPC is associated with a poor prognosis [12, 17, 18]

Methods

Results

Conclusion