Abstract 1698: Novel role of Tribbles 2 driving treatment-emergent neuroendocrine differentiation in prostate cancer

Abstract

Abstract Background: Prostate cancer initially responds to androgen receptor (AR) targeted therapies, such as enzalutamide. However, therapy-resistant, incurable disease invariably develops which remains as the major cause for mortality among prostate cancer patients. Molecular mechanism underlying emergence of resistant prostate cancer is yet to be fully characterized. Methods: We developed an in vitro cell culture model to mimic the clinical conditions in enzalutamide-resistant prostate cancer. These cells were transcriptionally profiled for gene expression analysis by Illumina Hi-Seq whole genome gene-expression array. Hits were validated by RT-qPCR and Western blot. For clinical correlation, enzalutamide treated prostate PDX models and surgical tumors (warm autopsy) from prostate cancer patients were comprehensively analyzed. Regulation and role of new hits were determined by treating cells and xenograft tumors with lentiviral-based shRNAs followed by analysis of cell viability, apoptosis, and effects on downstream targets. Pro-cancer effects were analyzed by cell proliferation, invasion, and soft-agar colony formation assays. Results: We identified that the Tribbles 2 (TRIB2) pseudokinase is overexpressed in enzalutamide-resistant prostate cancer (ERPC) cells. Elevated level of TRIB2 was also observed in enzalutamide treated PDX tumors, and in prostate tumors from patients who developed enzalutamide resistant metastatic disease. Interestingly, TRIB2 protein level is especially higher in prostate cancer cells and tumors, where the AR activity is low or absent due to genetic changes. Transfection and overexpression of TRIB2 results in aggressive prostate cancer cell growth and resistance to enzalutamide, apalutamide, darolutamide, and abiraterone. Furthermore, TRIB2 overexpression results in trans-differentiation in prostate cancer cells including neurite outgrowth and adeno-to-neuroendocrine lineage transition. Curiously, we found that TRIB2 downregulates luminal markers (AR, CK8), but upregulates the neuroendocrine markers (CHGA, NSE, SYP). Knockdown of TRIB2 triggers loss of neuroendocrine features, reduces viability of ERPC cells, and sensitize them back to enzalutamide, suggesting that TRIB2 plays a direct role in neuroendocrine differentiation and therapeutic resistance. We also found that TRIB2 upregulates the master neuronal transcription factor, BRN2, and the stemness factor, SOX2, and inhibition of either BRN2 or SOX2 re-sensitizes ERPC cells to enzalutamide. Conclusion: These findings reveal that TRIB2 is a novel driver for lineage plasticity and neuroendocrine differentiation in prostate cancer cells and greatly contributes to enzalutamide resistance. Citation Format: Jitender Monga, Craig Rogers, Shirish Gadgeel, Dhananjay Chitale, Jagadananda Ghosh. Novel role of Tribbles 2 driving treatment-emergent neuroendocrine differentiation in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1698.