Abstract PO021: Carcinoma-associated mesenchymal stem/stromal cells enhance ovarian cancer metastasis and increase cancer cell clonal heterogeneity through direct mitochondrial transfer
Authors: Pressimone Catherine A. , Frisbie Leonard G. , Pearson Alexander , Coffman Lan G. 
Published: March 2021

Abstract
Ovarian cancer is the most deadly gynecologic cancer largely due to early, diffuse metastatic spread. The ovarian cancer tumor microenvironment (TME) significantly impacts the metastatic capacity of ovarian cancer. We recently demonstrated that a critical stromal progenitor cell within the TME, termed carcinoma-associated mesenchymal stem/stromal cells (CA-MSCs), dramatically enhance the metastasis of ovarian cancer cells. CA-MSCs directly bind to cancer cells forming heterocellular units which co-metastasize. The goal of this work is to understand the mechanism driving CA-MSC-mediated ovarian cancer metastasis. We hypothesize that CA-MSCs increase the metastatic potential of ovarian cancer cells during the process of co-metastasis. Ovarian cancer uses both hematogenous and transcoelomic routes of metastasis therefore we used two ovarian cancer murine models to evaluate both modes of metastasis. We used a genomic barcode system to assess the clonal patterns of metastasis. Ovarian cancer cells with one unique barcode per cell were grown with or without CA-MSCs. Sites of metastasis were quantified and cancer cells were harvested from each site. Barcodes were sequenced to determined clonal representation at each metastatic site. We found CA-MSCs increased the number of metastatic sites in both the hematogenous and transcoelomic mouse models. Interestingly, CA-MSCs also increased the clonal heterogeneity at all metastatic sites (2.5 - 5 fold increase in blood, lung and liver). To investigate the mechanism enabling CA-MSC-mediated enhancement of clonal heterogeneity, we studied primary patient derived CA-MSCs and ovarian cancer cells. The process of metastasis poses unique metabolic stresses on cancer cells and normal mesenchymal stem cells are known to donate mitochondria to damaged epithelial cells. We therefore assessed if CA-MSCs use a similar mechanism to enhance survival of metastatic cancer cells. We stably labeled CA-MSC mitochondria with COX8-GFP via lentiviral transduction to enable the visualization and quantification of potential mitochondrial transfer. We demonstrate that CA-MSCs actively transfer mitochondria to cancer cells. This transfer is dependent on the physical interaction of CA-MSCs with cancer cells and is increased 2-4 fold when CA-MSCs and cancer cells form metastatic heterocellular units under non-adherent conditions. We tested the functional consequences of CA-MSC to cancer cell mitochondrial transfer and demonstrate cancer cells which receive mitochondria have increased chemotherapy resistance, increased sphere forming capacity and stem-like properties. Collectively, our results indicate CA-MSCs are a significant driver of ovarian cancer metastasis by forming heterocellular metastatic units which enable CA-MSC to cancer cell mitochondrial transfer. This leads to increased cancer cell survival and tumor initiation properties and represents a novel method of maintaining cancer cell heterogeneity during metastasis.
Citation Format: Catherine A. Pressimone, Leonard G. Frisbie, Alexander Pearson, Lan G. Coffman. Carcinoma-associated mesenchymal stem/stromal cells enhance ovarian cancer metastasis and increase cancer cell clonal heterogeneity through direct mitochondrial transfer [abstract]. In: Proceedings of the AACR Virtual Special Conference on the Evolving Tumor Microenvironment in Cancer Progression: Mechanisms and Emerging Therapeutic Opportunities; in association with the Tumor Microenvironment (TME) Working Group; 2021 Jan 11-12. Philadelphia (PA): AACR; Cancer Res 2021;81(5 Suppl):Abstract nr PO021.