Abstract PO-134: Identification of the cells of origin and tumor heterogeneity in neuroendocrine prostate cancer (NEPC) by single-cell analysis

Lineage plasticity has emerged as an important mechanism of therapeutic resistance in prostate cancer treated with a newer generation of AR signaling inhibitor (ARSi). As a consequence of lineage plasticity, treatment resistant prostate cancer loses epithelial cell identity, acquires stem cell-like properties and transforms into a neuroendocrine lineage. Loss of function mutations in Tp53, Rb1 and Pten are enriched in human neuroendocrine prostate cancer (NEPC). To understand the cell of origin and the molecular mechanism underlying lineage plasticity and NEPC development, we have used single-cell RNA-sequencing (scRNA-seq) technology to profile a genetically engineered mouse model with probasin-Cre driven Tp53, Rb1 and Pten deletions (referred to as TKO mouse) and Rb1 and Pten deletions (referred to as DKO mouse). We have profiled ~70,000 single cells from 16 mice of various ages and 6 additional mice that have undergone castration with or without testosterone addback. Our scRNA-seq analysis captures a developmental trajectory from luminal adenocarcinoma to neuroendocrine tumor, suggesting a newly discovered luminal cell type (L2) within the normal prostate may be the preferred cell of origin for NEPC development. Furthermore, combining scRNA-seq, flow cytometry and immunofluorescence analysis, we reveal tremendous heterogeneity within the neuroendocrine tumors with differential expression of several putative drivers, including SOX2, EZH2, AURKA, NMYC and POU2F3. Many of these genes have been previously implicated as drivers of NEPC development, while POU2F3 has been identified as a defining marker for a Tuft-variant small cell lung cancer (SCLC). Lastly, we have preliminary evidence that castration may have the potential to accelerate the transition from adenocarcinoma to NEPC, while adding back testosterone may delay the process in the TKO mice. Overall, in this study, we have identified a luminal L2 population as the putative preferred cell of origin for NEPC in the TKO model and revealed a previous under-appreciated heterogeneity within NEPC with differential expression of driver transcriptional, epigenetic and cell cycle regulators, which mirrors what has been described in the SCLC field. Citation Format: Jimmy L. Zhao, Samir Zaidi, Joseph Chan, Kristine Wadosky, Wouter Karthaus, Danielle Choi, Aura Agudelo Rivera, Anuradha Gopalan, Dana Rathkopf, Brett Carver, Wassim Abida, Howard Scher, Yu Chen, David Goodrich, Dana Pe’er, Charles L. Sawyers. Identification of the cells of origin and tumor heterogeneity in neuroendocrine prostate cancer (NEPC) by single-cell analysis [abstract]. In: Proceedings of the AACR Virtual Special Conference on Tumor Heterogeneity: From Single Cells to Clinical Impact; 2020 Sep 17-18. Philadelphia (PA): AACR; Cancer Res 2020;80(21 Suppl):Abstract nr PO-134.