CRISPR/Cas9 model of prostate cancer identifies Kmt2c deficiency as a metastatic driver by Odam/Cabs1 gene cluster expression

Prostate cancer (PCa) is the second leading cause of cancer-related deaths in men, and treatment options for metastatic disease remain limited. In this study, we employed Adeno-associated virus (AAV) delivery and CRISPR-Cas9 technology to simultaneously mutate multiple genes in the mouse prostate, in order to investigate positive genetic interactions during PCa progression. By targeting five tumor suppressor genes (Pten, Trp53, Rb1, Stk11, and RnaseL), we induced advanced prostate tumors in mice without metastasis, and mice reached humane endpoint at eight weeks. When three epigenetic factors (Kmt2c, Kmt2d, and Zbtb16) were further depleted, tumor progression remained identical, but metastases were observed in the lung of all mice. Hence, transcriptomic analysis was performed on 5g and 8g derived primary tumors (n = 4) and macro dissected lung metastases from 8g group (n = 4), using prostatic tissues from PBS-injected mice as control (n = 3). Libraries were prepared using an Illumina KAPA mRNA Hyperprep kit, followed by Illumina high-throughput sequencing on the NovaSeq 6000 platform.