Cancer-associated fibroblast-secreted glucosamine alters the androgen biosynthesis program in prostate cancer via HSD3B1 upregulation
Di Cui, Jianneng Li, Ziqi Zhu, Michael Berk, Aimalie Hardaway, Jeffrey McManus, Yoon-Mi Chung, Mohammad Alyamani, Shelley Valle, Ritika Tiwari, Bangmin Han, Maryam Goudarzi, Belinda Willard, Nima Sharifi
Di Cui, Jianneng Li, Ziqi Zhu, Michael Berk, Aimalie Hardaway, Jeffrey McManus, Yoon-Mi Chung, Mohammad Alyamani, Shelley Valle, Ritika Tiwari, Bangmin Han, Maryam Goudarzi, Belinda Willard, Nima Sharifi
View: Text | PDF
Research Article Oncology

Cancer-associated fibroblast-secreted glucosamine alters the androgen biosynthesis program in prostate cancer via HSD3B1 upregulation

Abstract

After androgen deprivation, prostate cancer frequently becomes castration resistant (CRPC), with intratumoral androgen production from extragonadal precursors that activate the androgen receptor pathway. 3β-Hydroxysteroid dehydrogenase-1 (3βHSD1) is the rate-limiting enzyme for extragonadal androgen synthesis, which together lead to CRPC. Here, we show that cancer-associated fibroblasts (CAFs) increased epithelial 3βHSD1 expression, induced androgen synthesis, activated the androgen receptor, and induced CRPC. Unbiased metabolomics revealed that CAF-secreted glucosamine specifically induced 3βHSD1. CAFs induced higher GlcNAcylation in cancer cells and elevated expression of the transcription factor Elk1, which induced higher 3βHSD1 expression and activity. Elk1 genetic ablation in cancer epithelial cells suppressed CAF-induced androgen biosynthesis in vivo. In patient samples, multiplex fluorescent imaging showed that tumor cells expressed more 3βHSD1 and Elk1 in CAF-enriched areas compared with CAF-deficient areas. Our findings suggest that CAF-secreted glucosamine increases GlcNAcylation in prostate cancer cells, promoting Elk1-induced HSD3B1 transcription, which upregulates de novo intratumoral androgen synthesis to overcome castration.

Authors

Di Cui, Jianneng Li, Ziqi Zhu, Michael Berk, Aimalie Hardaway, Jeffrey McManus, Yoon-Mi Chung, Mohammad Alyamani, Shelley Valle, Ritika Tiwari, Bangmin Han, Maryam Goudarzi, Belinda Willard, Nima Sharifi

×

Figure 1

CAFs increase the conversion from DHEA to active androgens in LNCaP cells by increasing 3βHSD1 expression and enzyme activity.

Options: View larger image (or click on image) Download as PowerPoint
CAFs increase the conversion from DHEA to active androgens in LNCaP cell...
(A) mRNA (by qPCR) and (B) protein expression of HSD3B1 and 3βHSD1 in LNCaP cells treated with CAF–conditioned medium (CAF-CM) for the indicated times. Gene and protein expression was normalized to RPLP0 and β-actin, respectively. (C) LNCaP cells were treated with CAF-CM for 48 hours followed by 100 nM DHEA for the indicated times. Downstream androgens in intracellular and media samples were quantitated by mass spectrometry. (D) LNCaP cells were treated with CAF-CM for 48 hours, followed by [3H]-DHEA (106 counts per minute) for the indicated times, followed by extraction of steroids from medium and quantitation by HPLC. (E) Mass spectrometry analysis of steroids in medium of LNCaP cells treated with DHEA along with CAF-CM derived from primary CAFs isolated from fresh prostate tumor tissue (patient no. 1). (F) Mass spectrometry analysis of primary CAF-CM from 3 patients with prostate cancer (patients 2, 3, and 4). (G) AR target gene (FKBP5 and TMPRSS2) mRNA expression in LNCaP cells (control or HSD3B1 siRNA; treated with 10nM DHEA and CAF-CM for 48 hours). Expression was normalized to untreated cells (data not shown), and RPLP0 was used as a loading control. (H) Cell viability of LNCaP control or HSD3B1 siRNA cells treated with DHEA along with control media or CAF-CM. Viability was normalized to the untreated control. Unless otherwise noted, data are shown as mean ± SEM. Significance was calculated using 2-tailed t tests or 1-way ANOVA as appropriate. *P < 0.05, **P < 0.01. Conc., Concentration.