Tumoroid model recreates clinically relevant phenotypes of high grade serous ovarian cancer (HGSC) cells, carcinoma associated fibroblasts, and macrophages

Tumoroid model recreates clinically relevant phenotypes of high grade serous ovarian cancer (HGSC) cells, carcinoma associated fibroblasts, and macrophages | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Tumoroid model recreates clinically relevant phenotypes of high grade serous ovarian cancer (HGSC) cells, carcinoma associated fibroblasts, and macrophages Kathleen Burkhard, Ayush Semwal, Ben Johnson, Kristina Chu, Riley Kranick, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6614892/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Ovarian cancer, the gynecological malignancy with the lowest survival rate, is significantly influenced by the tumor microenvironment. The mesenchymal subtype of high-grade serous carcinoma (HGSC) shows poor outcomes due to high stromal and low immune response. Single-cell RNA sequencing (scRNA-seq) of HGSC metastatic ascites has identified carcinoma-associated fibroblasts (CAFs), macrophages, and carcinoma-associated mesenchymal stem cells (CA-MSCs) as crucial drivers of immune exclusion, chemotherapy resistance, metastasis, and stem-like cell propagation. To explore this complex signaling, we developed heterogeneous tri-component tumoroids, incorporating HGSC cells (OVCAR3, OVCAR4, OVCAR8), primary MSCs, and U937-derived M2-like macrophages (M2-AAM) in defined ratios, each labeled with a fluorescent protein for distinct analysis. Upon a 48-hour treatment with carboplatin and/or paclitaxel, HGSC cells in tri-component tumoroids exhibited higher chemoresistance than HGSC-only spheroids. Flow cytometry revealed significant increases in cancer stem-like cell (CSC) markers CD44 and CD90 in the tri-component tumoroids. Conditioned medium from the tri-component tumoroids significantly enhanced HGSC cell migration compared to spheroids. Invasion assays further demonstrated that tri-component tumoroids penetrated monolayer of mCherry-labeled LP-9 mesothelial cells more effectively than spheroids. Additionally, scRNA-seq of tri-component tumoroids identified a unique cancer cell cluster enriched in epithelial-mesenchymal transition (EMT) and matrisome signatures, featuring a 14-gene signature linked to poor survival. MSCs in these tri-component tumoroids displayed a myofibroblastic-CAF signature, while macrophages indicated an ECM-associated and immunosuppressive phenotype. In conclusion, our 3D heterogenous tri-component tumoroids replicate key HGSC phenotypes, such as chemoresistance, CSC enrichment, migration, invasion, and EMT. This platform is invaluable for studying HGSC microenvironment interactions and preclinical testing of targeted therapies. Cell & Tissue Engineering Biomedical Engineering Cancer Biology Biomaterials Biotechnology and Bioengineering Obstetrics & Gynecology Oncology organoids tri-component tumoroids spheroids high grade serous ovarian cancer tumor microenvironment carcinoma-associated mesenchymal stem cells (MSC) (CA-MSC) tumor associated macrophages (TAM) alternately activated macrophages molecular subtyping scRNA-seq Full Text Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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