Stromal Cells Promote Matrix Deposition, Remodelling and an Immunosuppressive Tumour Microenvironment in a 3D Model of Colon Cancer

Abstract

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.

Temporal extracellular vesicle protein changes following intraarticular treatment with integrin α10β1-selected mesenchymal stem cells in equine osteoarthritis

“…It has been stated that serpin B9 has a cytoprotective function in MSCs ( 53 ). In other diseases such as colorectal cancer, the use of MSCs identified serpins as having immunomodulatory effects, acting on immune cells in order to induce a wound healing phenotype, as well as angiogenesis and epithelial to mesenchymal transition ( 54 ). Therefore, we hypothesize that the change in SF-EV proteome post MSC injection has the capacity to affect the serine endopeptidase pathway that is known for its detrimental effect on cartilage degradation and further animal studies are required to elucidate this.…”

Temporal Extracellular Vesicle Protein Changes following Intraarticular Treatment with Integrin α10β1-selected Mesenchymal Stem Cells in Equine Osteoarthritis

“…It has been stated that serpin B9 has a cytoprotective function in MSCs (35). In other diseases such as colorectal cancer, the use of MSCs identified serpins as having immunomodulatory effects, acting on immune cells in order to induce a wound healing phenotype, as well as angiogenesis and epithelial to mesenchymal transition (36). Therefore, we hypothesize that the change in SF-EV proteome post MSC injection has the capacity to affect the serine endopeptidase pathway that is known for its detrimental effect on cartilage degradation.…”

Investigating gene signatures associated with immunity in colon adenocarcinoma to predict the immunotherapy effectiveness using NFM and WGCNA algorithms

“…In addition to providing structural support, stromal cells, which include fibroblasts and vascular endothelial cells, control the recruitment and activation of immune cells through the secretion of cytokines [ 16 ]. In COAD, regulatory interactions between cytokines and immune cells may lead to the occurrence of immune escape, which promotes tumor cell proliferation and metastasis [ 17 ]. In the microenvironment of CRC, IL6 generated by CD163 + tumor-associated macrophages stimulates epithelial mesenchymal transition by controlling the STAT3/miR-506-3p/FoxQ1 pathway, which in turn promotes CRC cell invasion and migration [ 18 ].…”