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T Cell Gene Editing Using CRISPR/Cas9 and Experimental Kidney Ischemia-Reperfusion Injury. | LitMetric

T cells play pathophysiologic roles in kidney ischemia-reperfusion injury (IRI), and the nuclear factor erythroid 2-related factor 2/kelch-like ECH-associated protein 1 (Nrf2/Keap1) pathway regulates T cell responses. We hypothesized that clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated -knockout (KO) augments Nrf2 antioxidant potential of CD4+ T cells, and that -KO CD4+ T cell immunotherapy protects from kidney IRI. CD4+ T cell -KO resulted in significant increase of Nrf2 target genes NAD(P)H quinone dehydrogenase 1, heme oxygenase 1, glutamate-cysteine ligase catalytic subunit, and glutamate-cysteine ligase modifier subunit. -KO cells displayed no signs of exhaustion, and had significantly lower levels of interleukin 2 (IL2) and IL6 in normoxic conditions, but increased interferon gamma in hypoxic conditions . , adoptive transfer of -KO CD4+ T cells before IRI improved kidney function in T cell-deficient mice compared with mice receiving unedited control CD4+ T cells. -KO CD4+ T cells isolated from recipient kidneys 24 h post IR were less activated compared with unedited CD4+ T cells, isolated from control kidneys. Editing Nrf2/Keap1 pathway in murine T cells using CRISPR/Cas9 is an innovative and promising immunotherapy approach for kidney IRI and possibly other solid organ IRI. CRISPR/Cas9-mediated -KO increased Nrf2-regulated antioxidant gene expression in murine CD4+ T cells, modified responses to hypoxia and kidney IRI. Gene editing targeting the Nrf2/Keap1 pathway in T cells is a promising approach for immune-mediated kidney diseases.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10171956PMC
http://dx.doi.org/10.1089/ars.2022.0058DOI Listing

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