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Crosstalk of glial cells and other components of the tumor microenvironment

ABSTRACT: The crosstalk of glial cells and other components of the tumor microenvironment is a complex process, suggested to exacerbate tumor development. Here, we treated murine primary enteric glial cells (EGCs) with IL-1β, a cytokine that has been shown to be upregulated in many solid tumors. Mass spectrometry was performed to detect the factors secreted by glial cells. Diverse proteins were found differentially regulated upon IL-1β treatment.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Marc Sylvester

LAB HEAD: Prof. Dr. rer. nat. Sven Wehner

PROVIDER: PXD045911 | Pride | 2024-06-05

REPOSITORIES: Pride

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			Action	DRS
	221202_P-444-BR_020-025_tr2_221205.mzML	Mzml
	221202_P-444-BR_020-025_tr2_221205.mzid.gz	Mzid
	221202_P-444-BR_020-025_tr2_221205.pdResult	Other
	221202_P-444-BR_020-025_tr2_221205.pdResultView	Other
	221202_P-444-BR_020_tr2.raw	Raw

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Project description:Enteric glial cells are important players in the control of motility, intestinal barrier integrity and inflammation. During inflammation, they switch into a reactive phenotype enabling them to release inflammatory mediators, thereby shaping the inflammatory environment. While a plethora of well-established in vivo models exist, cell culture models necessary to decipher the mechanistic pathways of enteric glial reactivity are less well standardized. In particular, the composition of extracellular matrices (ECM) can massively affect the experimental outcome. Considering the growing number of studies involving primary enteric glial cells, a better understanding of their homeostatic and inflammatory in vitro culture conditions is needed. We examined the impact of different ECMs on enteric glial culture homeostasis and immune responsiveness. We used 3’ bulk mRNA sequencing to compare cultured cells with in vivo enteric glial transcriptomes isolated from Sox10iCreERT2Rpl22HA/+ mice. Transcriptional analysis revealed strong similarities between enteric glia on Matrigel and laminin with enrichment of gene sets supporting neuronal differentiation, while cells on poly-L-ornithine showed enrichment related to cell proliferation. Comparing cultured and in vivo enteric glial transcriptomes revealed a 50% overlap. IL-1β treatment showed distinct coating-dependent gene expression signatures, with an enrichment of genes related to myeloid and epithelial cell differentiation on Matrigel and laminin coatings, while poly-L-ornithine induced more gene sets related to lymphocyte differentiation. Together, changes in differentiation and immune activation of primary enteric glial cells proved a strong effect of the ECM. We identified Matrigel and laminin as pre-eminent substrates for murine enteric glial cultures. These new insights will help to standardize and improve enteric glial culture quality and reproducibility between studies in the future, allowing a better comparison of their functional role in enteric neuroinflammation.

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Crosstalk of glial cells and other components of the tumor microenvironment

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