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Single-cell RNA-sequencing of KPPF;Col1smaKO mouse models with pancreatic tumors

ABSTRACT: We employed dual-recombinase genetic mouse models of spontaneous PDAC mice (KPPF;Col1smaKO) to delete Col1 (type I collagen) specifically in myofibroblasts, in comparison with control KPPF mice. Single-cell RNA-sequencing analyses were performed on unfractionated live cell mixtures from pancreatic tumors of KPPF mice and KPPF;Col1smaKO mice, to investigate the impact of myofibroblast-specific Col1 deletion on tumor microenvironment.

ORGANISM(S): Mus musculus

PROVIDER: GSE166298 | GEO | 2021/02/07

REPOSITORIES: GEO

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Project description:Pericryptal myofibroblasts in the colon and rectum play an important role in regulating the normal colorectal stem cell niche and facilitating tumour progression. Myofibroblasts have previously mostly been distinguished from normal fibroblasts only by the expression of Î± smooth muscle actin (Î±SMA). We now identify AOC3, a surface monoamine oxidase, as a new marker of myofibroblasts by showing that it is the target protein of the myofibroblast reacting monoclonal antibody (mAb), PR2D3. The normal and tumour tissue distribution and the cell line reactivity of AOC3 match that expected for myofibroblasts. We have shown that the surface expression of AOC3 is sensitive to digestion by trypsin and collagenase and that anti-AOC3 antibodies can be used for FACS sorting of myofibroblasts obtained by non-enzymatic procedures. Whole genome microarray mRNA expression profiles of myofibroblasts and skin fibroblasts revealed four additional genes that are significantly expressed differentially between these two cell types; NKX2-3 and LRRC17 are expressed in myofibroblasts and SHOX2 and TBX5 in skin fibroblasts. Transforming Growth Factor Î² (TGFÎ²) substantially down-regulated AOC3 expression in myofibroblasts but not in skin fibroblasts, in which it dramatically increased the expression of Î±SMA. A knockdown of NKX2-3 in myofibroblasts caused a decrease of myofibroblast-related gene expression and an increased expression of the fibroblast associated gene, SHOX2, suggesting that NKX2-3 is a key mediator for maintaining myofibroblast characteristics. Our results show that colorectal myofibroblasts, as defined by the expression of AOC3, NKX2-3 and other markers, are a distinctly different cell type from TGFÎ² activated fibroblasts. colorectal myofibroblast specific markers and expression profiles were sought by comparing four primary myofibroblast cultures to a panel of four dermal and foreskin fibroblast cell lines Four primary myofibroblast cultures established from adult human colon compared to four skin fibroblast cell lines to identify intestinal myofibroblast specific markers

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Single-cell RNA-sequencing of KPPF;Col1smaKO mouse models with pancreatic tumors

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