Cancer-associated fibroblast-secreted glucosamine alters the androgen biosynthesis program in prostate cancer via HSD3B1 upregulation
Di Cui, … , Belinda Willard, Nima Sharifi
Di Cui, … , Belinda Willard, Nima Sharifi
Published April 3, 2023
Citation Information: J Clin Invest. 2023;133(7):e161913. https://doi.org/10.1172/JCI161913.
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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

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Figure 3

O-GlcNAcylation after CAF-CM treatment is attributable to glucosamine and induces 3βHSD1 expression and enzyme activity.

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O-GlcNAcylation after CAF-CM treatment is attributable to glucosamine an...
(A) Western blot analysis of O-GlcNAcylated proteins in LNCaP and C42 cells treated with CAF–conditioned medium (CAF-CM) (left) or increasing concentrations of glucosamine (right) for 48 hours. (B) Western blot analysis of O-GlcNAcylated proteins, OGT, and 3βHSD1 protein expression in LNCaP cells expressing 2 shRNAs targeting OGT (OCG200 and OGT3652) or scrambled shRNA and treated with CAF-CM or 10 mM glucosamine for 48 hours. (C) O-GlcNAcylated proteins, OGT, and 3βHSD1 protein expression in C4-2 cells treated with CAF-CM or 10 mM glucosamine in the presence of DMSO (vehicle control) or 75 μM OSMI-1, an OGT inhibitor. (D) HPLC analysis of DHEA metabolism in LNCaP cells expressing scrambled shRNA, shOCG200, and shOGT3652 treated with CAF-CM or 10 mM glucosamine for 48 hours, followed by addition of [3H]-DHEA (100 nM) for the indicated times. Significance was calculated using 1-way ANOVA. (E) Western blot analysis of OGA, O-GlcNAcylated proteins, and 3βHSD1 expression in LNCaP (left) and C4-2 (right) cells transduced with control (sgControl) or sgRNA targeting OGA (sgOGA) after 48-hour treatment with CAF-CM or 10 mM glucosamine. (F) Pearson correlation analysis of HSD3B1 and OGA mRNA expression in prostate cancer (MSKCC Prostate Oncogenome Project, GSE21032). (G) HPLC analysis of DHEA metabolism in media of LNCaP control or OGA-KO cells treated with CAF-CM or glucosamine for 48 hours, followed by [3H]-DHEA (100 nM) for the indicated times; data were normalized to untreated control. (H) Gene expression of AR target genes PSA, FKBP5, and TMPRSS2 in control and OGA-KO LNCaP cells treated with CAF-CM or 10 mM glucosamine plus 10 nM DHEA for 48 hours. Significance was calculated using 2-tailed t tests. *P < 0.05, **P < 0.01.