1493 Developing a 3D multicellular model to investigate stromal and immune interactions in colorectal cancer tumour microenvironment

作者：Eileen Reidy, Niamh Leonard, Abhay Pandit, Aideen E. Ryan · 发表于：Regular and Young Investigator Award Abstracts · 年份：2023 · DOI：10.1136/jitc-2023-sitc2023.1493 · 被引用次数：1 · 研究领域：Colorectal Cancer Treatments and Studies、Colorectal Cancer Surgical Treatments、Genetic factors in colorectal cancer

Background Colorectal cancer (CRC) is the 3rd most common cause of cancer related deaths worldwide. 1 Patients with stromal dense tumours (CMS4) have the worst disease-free progression survival rates and account for 23% of all CRC patients. 2 These patients have an inflamed immunophenotype and often do not respond well to current treatment options. Understanding cell-cell and cell-ECM interactions in CMS4 is necessary for developing new treatments for CMS4 patients. 3 We aim to develop a 3D model which replicates aspects of CMS4 CRC tumour microenvironment (TME) in order to study interactions taking place between colorectal cancer cells, stroma and immune cells in the TME of CRC. Methods Spheroids were established from HCT116 and HT29 human CRC cell lines, and embedded into collagen type 1 hydrogels. To recapitulate stromal dense tumour microenvironments, bone marrow derived hMSCs were incorporated in the spheroids. Viability was analysed using calcein AM and propidium iodide staining. Using confocal microscopy and imageJ the extra cellular matrix of the spheroids was analysed. Finally, to mimic the immune landscape of CMS4 CRC patients, spheroids were co-cultured with Jurkat cells (T cell line) or activated T cells isolated from healthy patients. Using flow cytometry immune cell activation and polarisation in the 3D model was assessed. Results Incorporating MSCs leads to reduced cell death and increased outgrowth from spheroids, mimicking the increased metastatic capa...