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 3

3βHSD1 enhances DNA repair in irradiated PCa cells treated with DHEA.

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3βHSD1 enhances DNA repair in irradiated PCa cells treated with DHEA. (A...
(A) Representative immunofluorescence images (top) and quantitation (bottom) of phospho-γH2AX foci in LNCaP cells expressing shControl or shHSD3B1 pretreated with 50 nM DHEA for 48 hours followed by 0 Gy or 4 Gy irradiation. The γH2AX foci were quantified as foci per nucleus at each time point. All data are represented as mean values ± 95% CI (P values were calculated using 2-tailed t test). (B) Representative immunofluorescence images (top) and quantification (bottom) of γH2AX foci in LAPC4 cells stably expressing HSD3B1 1245A, 1245C, or EV control. The cells were treated for 48 hours with DHEA, followed by 0 Gy or 4 Gy irradiation. All data are represented as mean values ± 95% CI (P values were calculated using 2-way ANOVA with Bonferroni’s multiple comparison test). (C) Neutral COMET assay and tail moment quantitation of LNCaP and (D) C4-2 cells expressing shControl and shHSD3B1 following pretreatment with DHEA and irradiation (0 Gy and 4 Gy). All data are represented as mean values ± 95% CI (P values were calculated using 2-tailed t test) (47). *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars: 5 μm. (E) Immunoblot of DDR markers 12 hours after irradiation in LNCaP cells pretreated with 50 nM DHEA. Lamin B1 was used as a loading control. (F) Immunoblot analysis of DDR markers 12 hours after irradiation in LAPC4 expressing HSD3B1 (1245A, 1245C, or EV control) pretreated with 50 nM DHEA. (G) Volcano plots depicting differentially expressed genes in HSD3B1 (1245A) LAPC4 cells (left) and HSD3B1 (1245C) LAPC4 cells (right) compared with the EV control. Key DDR genes are highlighted in black.