Next-generation sequencing to define the impact of Cxcl1 on murine MSC transcriptome and BM-PCa tumor microenvironment

Purpose: Prostate cancer most frequently metastasizes to bone and is incurable. Metastatic prostate cancer cells thrive in bone through molecular regulation of surrounding bone stroma, including upregulation of CXCL8 (CXCL1 in mice) in MSCs. This study characterizes the changes in the bone marrow mesenchymal stem cells (MSCs) upon targeted deletion of Cxcl1 in murine MSC. It also characterized the changes in the tumor/stromal cells of tumors with Cxcl1 KO MSCs or control MSCs. These studies seek to determine the importance of MSC-derived CXCL1 in bone-metastatic prostate cancer. Overall design: Cxcl1 was knocked down in murine bone marrow MSCs using stable lentiviral infection of Cxcl1-targeted or scrambled control shRNAs. RNA was collected for downstream bulk RNA-seq analyses. CRSIPR-Cas9 was used to generate Cxcl1 KO murine bone marrow MSCs. RNA was collected using Trizol reagent for downstream bulk RNA-seq analyses of Cxcl1 KO MSC and control MSC cell lines as well as tumor/stromal cells from an intratibial RM1 mouse prostate cancer model containing either Cxcl1 KO MSCs or control MSCs.