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Learn MoreThere is limited understanding of how immune-mediated damage impacts intestinal stem cells (ISCs). We found that stem cell compartment injury is a shared feature of both alloreactive and autoreactive intestinal immunopathology, reducing ISCs and impairing their recovery in T-cell-mediated injury models. Modeling with ex vivo epithelial cultures indicated ISC depletion and impaired human as well as murine organoid survival upon co-culture with activated T cells, and screening of effector pathways identified Interferon- (IFN) as a principal mediator of ISC compartment damage. Flow cytometry analysis confirmed expression of IFN receptor on ISCs. To investigate if the receptor was functional, we performed transcriptome analysis of ISCs treated with IFN. In brief, mRNA profiles of sorted Lgr5-high cells treated with/without IFN were generated by deep sequencing, in triplicate, using the TruSeq SBS Kit V4 (Illumina). RNA profiles confirmed a robust IFN transcriptional signature in the stem cells, as several IFN-related genes were upregulated shortly after exposure, indicating direct activity of IFN in ISCs. Epithelial cultures with IFN-receptor-deficient ISCs and purified stem cell colonies also indicated direct targeting of the ISCs. This study showed that dysregulated T cell activation and Interferon- production are potent mediators of ISC injury. SOURCE: Bic MSKCC (bic-geo-submission@cbio.mskcc.org) - Memorial SLoan-Kettering Cancer Center
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