Intratumoral androgen biosynthesis associated with 3β-hydroxysteroid dehydrogenase 1 promotes resistance to radiotherapy in prostate cancer
Shinjini Ganguly, … , Nima Sharifi, Omar Y. Mian
Shinjini Ganguly, … , Nima Sharifi, Omar Y. Mian
Published November 15, 2023
Citation Information: J Clin Invest. 2023;133(22):e165718. https://doi.org/10.1172/JCI165718.
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Research Article Endocrinology

Intratumoral androgen biosynthesis associated with 3β-hydroxysteroid dehydrogenase 1 promotes resistance to radiotherapy in prostate cancer

Abstract

Half of all men with advanced prostate cancer (PCa) inherit at least 1 copy of an adrenal-permissive HSD3B1 (1245C) allele, which increases levels of 3β-hydroxysteroid dehydrogenase 1 (3βHSD1) and promotes intracellular androgen biosynthesis. Germline inheritance of the adrenally permissive allele confers worse outcomes in men with advanced PCa. We investigated whether HSD3B1 (1245C) drives resistance to combined androgen deprivation and radiotherapy. Adrenally permissive 3βHSD1 enhanced resistance to radiotherapy in PCa cell lines and xenograft models engineered to mimic the human adrenal/gonadal axis during androgen deprivation. The allele-specific effects on radiosensitivity were dependent on availability of DHEA, the substrate for 3βHSD1. In lines expressing the HSD3B1 (1245C) allele, enhanced expression of DNA damage response (DDR) genes and more rapid DNA double-strand break (DSB) resolution were observed. A correlation between androgen receptor (AR) expression and increased DDR gene expression was confirmed in 680 radical prostatectomy specimens. Treatment with the nonsteroidal antiandrogen enzalutamide reversed the resistant phenotype of HSD3B1 (1245C) PCa in vitro and in vivo. In conclusion, 3βHSD1 promotes prostate cancer resistance to combined androgen deprivation and radiotherapy by upregulating DNA DSB repair. This work supports prospective validation of early combined androgen blockade for high-risk men harboring the HSD3B1 (1245C) allele.

Authors

Shinjini Ganguly, Zaeem Lone, Andrew Muskara, Jarrell Imamura, Aimalie Hardaway, Mona Patel, Mike Berk, Timothy D. Smile, Elai Davicioni, Kevin L. Stephans, Jay Ciezki, Christopher J. Weight, Shilpa Gupta, Chandana A. Reddy, Rahul D. Tendulkar, Abhishek A. Chakraborty, Eric A. Klein, Nima Sharifi, Omar Y. Mian

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Figure 1

Loss of 3βHSD1 expression reduces colony formation and cell survival in irradiated PCa cells.

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Loss of 3βHSD1 expression reduces colony formation and cell survival in ...
HSD3B1 mRNA (top panels) and 3βHSD1 protein expression (bottom panels) in (A) LNCaP, C4-2, and VCaP cells stably expressing shRNA targeting HSD3B1 (shHSD3B1) or nonsilencing shRNA (shControl). Gene expression was normalized to ACTB, and GAPDH was used as a loading control for immunoblotting. All data are represented as mean ± 95% CI from triplicates of 2 independent experiments (unpaired 2-tailed t test) (B) LAPC4 and RWPE-1 cells stably expressing dox-inducible restrictive (1245A) or permissive (1245C) HSD3B1. Gene expression of HSD3B1 (top panels) was assessed by qPCR (normalized to ACTB). All data are represented as mean ± SEM from triplicates of 2 independent experiments (1-way ANOVA with Bonferroni’s multiple-comparison test), and protein levels of 3βHSD1 (bottom panels) were measured by immunoblotting (normalized to β-actin). (C) Surviving cell fraction and (D) colony formation assay of LNCaP cells expressing shHSD3B1 or shControl treated with 4 or 8 Gy radiation and cultured for 14 days in csFBS medium containing ethanol (top panel), 50 nM DHEA (middle panel), or 1 nM R1881 (bottom panel) followed by crystal violet staining. Representative images of colonies formed after treatment with 0, 4, or 8 Gy radiation and cultured in the presence or absence of androgens. Original magnification, ×2. (E) The number of viable RWPE1 colonies stably expressing 1245A or 1245C HSD3B1 treated with 0, 4, or 8 Gy radiation and cultured in csFBS media containing ethanol (top panel), 50 nM DHEA (middle panel), or 1 nM R1881 (bottom panel). All data are represented as mean values ± 95% CI from triplicates in 2 independent experiments (unpaired 2 tailed t test). *P < 0.05; **P < 0.01; ***P < 0.001.