Background: Age-adjusted incidence and mortality rates for prostate cancer (PCa) among African American (AA) men are 1.6- and 2.4-fold greater, respectively, than among white men. The more aggressive characteristics of AA PCa account for a significant component of the PCa disparity, in addition to social determinants of health. Differences in androgenic activities in AA versus white populations and PCa patients have been observed and, clinically, AAs have a less complete response to androgen deprivation than whites do. One of the critical androgen receptor signaling targets in PCa is AR-V7, an androgen receptor variant that lacks the ligand-binding domain, is constitutively active, and associates with castration-resistant PCa, poorer clinical outcomes, and resistance to androgen ablation/androgen receptor inhibition therapies. This work addresses the urgent need to develop a novel therapeutic strategy capable of inhibiting AR-V7.
Methods: We have designed and synthesized a novel chemically modified splice switching oligonucleotide (SSO) to correct aberrant splicing leading to production of AR-V7 as well as a control scrambled SSO. After transfection of PCa cell lines derived from AA and white patients with these SSOs, we have examined AR-V7 as well as androgen receptor RNA and protein levels using qPCR and Western blot analysis, respectively. Resulting alterations in proliferation have been assessed by monitoring cell growth using an Incucyte Live-Cell Imaging System and associated software.
Results: Transfection of a panel of PCa cell lines derived from AA and white patients with AR-V7 SSO decreases AR-V7 RNA and protein in a dose-dependent manner while maintaining expression levels of full-length androgen receptor. Preliminary data suggest this biochemical response correlates with a biologically significant phenotype. AR-V7 SSO decreases proliferation in PCa cells, including proliferation in the presence of the androgen receptor inhibitor, enzalutamide, over and above the reduction seen in response to enzalutamide alone in both enzalutamide-sensitive and -resistant PCa cell lines. Additionally, transfection of PCa cells with SSOs designed to target nearby splicing enhancer sequences partially reduces AR-V7 RNA expression. Further studies to examine the effects of this SSO on transactivation activity, signaling and PCa cell biology, and the therapeutic efficacy of this SSO in AA and white PCa patient-derived primary cell lines and xenografts are under way.
Conclusions: These studies suggest that a SSO can be developed to modulate androgen receptor signaling, inhibiting constitutive signaling and restoring ligand dependency, with interference of nearby splicing enhancer sequences contributing to its function. Such an SSO could represent a novel therapeutic strategy with the potential to reduce PCa disparities for AA men and improve outcomes for men of all races with aggressive disease driven by this mechanism.
Citation Format: Bonnie L. LaCroix, Brendon M. Patierno, Bruce A. Sullenger, Daniel J. George, Steven R. Patierno, Jennifer A. Freedman. Development of a novel therapeutic splice-switching oligonucleotide targeting race-related androgen receptor signaling and aggressive prostate cancer [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr B006.