mRNA sequencing to elucidate monocyte differentiation potential in OV90, OVCAR3, OVCAR4, OVAR5, and OVCA432 ovarian cancer cell lines

Alternatively-activated macrophages (AAMs), an anti-inflammatory macrophage subpopulation, have been implicated in the progression of high grade serous ovarian carcinoma (HGSOC). Increased levels of AAMs are correlated with poor HGSOC survival rates, and AAMs increase the attachment and spread of HGSOC cells in vitro. However, the mechanism by which monocytes in the HGSOC tumor microenvironmentare differentiated and polarized to AAMs remains unknown. Using an in vitro co-culture device, we cultured naive, primary human monocytes with a panel of five HGSOC cell lines over the course of seven days. An empirical Bayesian statistical method, EBSeq, was used to couple RNA-seq with observed monocyte-derived cell phenotype to explore which HGSOC-derived soluble factors supported differentiation to CD68+ macrophages and subsequent polarization towards CD163+ AAMs. Overall design: RNA-sequencing was performed on five (OV90, OVCAR3, OVCAR4, OVCAR5, OVCA432) immortalized HGSOC cell lines grown in monoculture. Significant changes in gene expression were correlated to AAM induction potential to identify soluble factors with potential contributions to HGSOC-mediated monocyte differentiation.