Abstract PR016: Carcinoma-associated mesenchymal stem cells promote ovarian cancer metastasis by increasing tumor heterogeneity through direct mitochondrial transfer
Authors: Frisbie Leonard G. , Pressimone Catherine A. , Atiya Huda I. , Pearson Alexander T. , Coffman Lan G. 
Published: January 2023

Abstract
Ovarian cancer is characterized by early, diffuse metastatic spread with most women presenting with widespread abdominal metastasis at the time of diagnosis. Prior work demonstrates carcinoma-associated mesenchymal stem cells (CA-MSCs) enhance ovarian cancer metastasis through a process of direct cellular interaction and formation of heterocellular CA-MSC and tumor cell complexes. Here we demonstrate CA-MSCs enhance metastasis via increasing tumor cell heterogeneity through mitochondrial donation. CA-MSCs directly interact with ovarian cancer cells forming tunneling nanotubules (TNTs). CA-MSCs transfer live mitochondria via these TNTs to ovarian cancer cells. This mitochondrial donation preferentially occurs in ovarian cancer cells with the least endogenous mitochondria (‘mito poor’ cancer cells). Mito poor cancer cells demonstrate decreased proliferation, increased sensitivity to chemotherapy and decreased oxidative phosphorylation compared to ‘mito rich’ cancer cells. CA-MSCs rescue the phenotype of these mito poor cancer cells restoring their proliferative capacity, increasing chemotherapy resistance and increasing oxidative phosphorylation. We validated these findings in a fully autologous system using CA-MSCs and cancer cells derived from the same patient to prevent confounding effects of cellular response to foreign organelle/DNA. Using a knockdown of the mitochondrial motor protein, MIRO1, we demonstrate mitochondrial transfer is necessary for the CA-MSC-mediated rescue of mito poor cancer cells. We developed a haplotype-specific quantification of mitochondrial DNA to differentiate CA-MSC derived mitochondria from endogenous tumor cell mitochondria. We used this system to both quantify the amount of CA-MSC mitochondrial donation and to demonstrate CA-MSC donated mitochondria persist in tumor cells over at least 14 days. Importantly, CA-MSC mitochondrial donation occurs in vivo and is associated with decreased survival in an orthotopic ovarian cancer mouse model. A DNA barcoding system was used to quantify tumor cell clonal heterogeneity. Using this system, we demonstrate CA-MSCs significantly enhance tumor cell heterogeneity, particularly during metastasis. This increase in tumor cell heterogeneity is dependent on CA-MSC mitochondrial transfer. Collectively, we report CA-MSC mitochondrial transfer as a critical mediator of ovarian cancer survival, heterogeneity and metastasis representing a potentially powerful therapeutic target.
Citation Format: Leonard G. Frisbie, Catherine A. Pressimone, Huda I. Atiya, Alexander T. Pearson, Lan G. Coffman. Carcinoma-associated mesenchymal stem cells promote ovarian cancer metastasis by increasing tumor heterogeneity through direct mitochondrial transfer [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr PR016.