Abstract
Simple SummaryColorectal cancer is the third most common type of cancer in the world. Immune cells and normal supporting cells (MSCs) within a tumour affect patient survival and change how well treatments work. This research aimed to develop a more relevant 3D cancer model that combines MSCs and immune cells with cancer cells to test the effects of multiple cell types on tumour growth. We successfully developed a 3D model that shows that MSCs and immune cells can change the cancer-supporting environment around the tumour cells. We show that combining MSCs and immune cells with cancer cells can increase the level of immune-suppressing molecules they release and change immunotherapeutic drug targets on the cancer cells, similar to changes seen in human tumours. Using this 3D model for research may be better for testing new drugs than traditional 2D methods and could enable the identification of new drug targets.Colorectal cancer (CRC) is the third leading cause of cancer-related deaths worldwide. CRC develops in a complex tumour microenvironment (TME) with both mesenchymal stromal cells (MSCs) and immune infiltrate, shown to alter disease progression and treatment response. We hypothesised that an accessible, affordable model of CRC that combines multiple cell types will improve research translation to the clinic and enable the identification of novel therapeutic targets. A viable gelatine-methacrloyl-based hydrogel culture system that incorporates CRC cells with MSCs and a monocyte cell line was developed. Gels were analysed on day 10 by PCR, cytokine array, microscopy and flow cytometry. The addition of stromal cells increased transcription of matrix remodelling proteins FN1 and MMP9, induced release of tumour-promoting immune molecules MIF, Serpin E1, CXCL1, IL-8 and CXCL12 and altered cancer cell expression of immunotherapeutic targets EGFR, CD47 and PD-L1. Treatment with PD153035, an EGFR inhibitor, revealed altered CRC expression of PD-L1 but only in gels lacking MSCs. We established a viable 3D model of CRC that combined cancer cells, MSCs and monocytic cells that can be used to research the role the stroma plays in the TME, identify novel therapeutic targets and improve the transitional efficacy of therapies.
Highlights
Colorectal cancer (CRC) is the third most common cancer worldwide and remains a significant health challenge, with over 1.8 million people diagnosed annually [1]
Our results show that the GelMA 3D platform supports the growth of primary mesenchymal stromal cells (MSCs) and immune cells, which can be very sensitive to their culture system, while allowing the formation and proliferation of tumour spheroids
Our model reveals that EGFR expression is increased on CRC when cultured with stromal cells
Summary
Colorectal cancer (CRC) is the third most common cancer worldwide and remains a significant health challenge, with over 1.8 million people diagnosed annually [1]. MSCs encompass a variety of cell types of mesenchymal origin with similar cell phenotypes, such as myo-fibroblasts and cancerassociated fibroblasts (CAFs), and they can be resident cells or recruited to the tumour site to promote progression [7,8]. Immune cells, both adaptive and innate, are found in the CRC TME, and depending on their phenotype and location, have different effects on tumour growth and development [9,10,11]. Classification of CRC tumours based on their stromal signature may help predict treatment response and disease-free survival rates [12]
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