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
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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 4

High O-GlcNAcylation increases Elk1 to induce 3βHSD1 expression and enzyme activity.

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High O-GlcNAcylation increases Elk1 to induce 3βHSD1 expression and enzy...
(A) Protein expression of ELK1 in LNCaP and C42 cells treated with CAF–conditioned medium (CAF-CM) (left) and increasing concentrations of glucosamine (right) for 48 hours. (B) ELK1 protein expression in LNCaP cells expressing control or OGT shRNA (left) and sgRNA (right) treated with CAF-CM or 10 mM glucosamine for 48 hours. (C) Pearson correlation analysis of ELK1 and OGA mRNA expression in prostate cancer (GSE21032). (D) Left: Gene expression of HSD3B1 in control (sgControl) or ELK1-KO (sgELK1) LNCaP cells treated with CAF-CM or 10 mM glucosamine for 48 hours. Right: Luciferase assay of LNCaP control and ELK1-KO cells cotransfected with an HSD3B1 promoter-firefly luciferase and Renilla luciferase plasmid constructs, which were treated with CAF-CM or 10 mM glucosamine 48 hours. (E) ChIP assay of Elk1. C4-2 cells were treated with 10 mM glucosamine for 24 hours. (F) Protein expression of ELK1 and 3βHSD1 in sgControl and ELK1-KO LNCaP cells treated with CAF-CM or glucosamine. (G) HPLC analysis of steroids in media of LNCaP cells expressing sgControl and ELK1 KO (left) or siRNA and shRNA knockdown of ELK1 (right). Cells were treated with CAF-CM or glucosamine for 48 hours, followed by [3H]-DHEA (100 nM) for 48 hours. (H) Gene expression of PSA, TMPRSS2, and FKBP5 in sgControl and ELK1-KO LNCaP cells treated with 10 nM DHEA along with CAF-CM or glucosamine for 48 hours. (I) Pearson correlation analysis of ELK1 and HSD3B1 mRNA expression in prostate cancer (GSE21032). Significance was calculated using 2-tailed t tests. *P < 0.05, **P < 0.01.