Single-cell Analysis Of Cell Differentiation And Origin Of Human Neuroendocrine Prostate Cancer Cells

Background: Neuroendocrine prostate cancer(NEPC)is one of the most aggressive subtypes of castration-resistant prostate cancer(CRPC).Although much progress has been made in understanding the development of neuroendocrine prostate cancer,the cellular architecture associated with neuroendocrine differentiation(NED)in human prostate cancer remains incompletely understood.Single-cell transcriptome analysis can provide new insights into the cellular phenotypes/states associated with NED in prostate cancer.Objects: Firstly,single cell sequencing analysis was used to reveal intratumoral and intertumoral heterogeneity of NEPC,and to characterize the comprehensive molecular phenotypes of NED cells;Secondly,divergent algorithms and patient cohorts were exploited to figure out the source of NED cells.Finally,by studying differentially expressed genes in NED cells,new potential drug targets were explored for NEPC treatment.Materials and methods: Single-cell RNA-Seq(sc RNA-Seq)analysis was performed on 11 biopsied tissues from 9 patients with castrationresistant prostate cancer(CRPC)using a droplet-based platform.Prostate cancer tissue microarray(PC TMA)analyses of 297 patients were used to determine general cellular phenotypes identified in the single-cell data.Bulk transcriptional datasets were used to determine the robustness of single-cell dataset-derived NED-related genes.Results: we obtained a total of 60,942 high-quality cells that were obtained from needle biopsies of 9 castration-resistant prostate cancer(CRPC)patients.Our analysis identified NED in samples from 4 patients,including two pure NED tumors and two mixed adenocarcinoma-NE tumors.All identified neuroendocrine(NE)cells were transcriptionally similar to luminal-like malignant cells.In two mixed adenocarcinoma-NE tumors,lineage trajectory analysis suggests that focal neuroendocrine differentiation exclusively originates from luminal-like malignant cells rather than the basal compartment.PC TMAs validated the generality of the luminal phenotype of NE cells.Conclusions: In summary,our single-cell study provides direct evidence for the cellular states underlying neuroendocrine transdifferentiation of human prostate cancer cells and reveals that a luminal epithelial state is highly linked to NED of prostate cancer cells.