T Cell Nrf2/Keap1 Gene Editing Using CRISPR/Cas9 and Experimental Kidney Ischemia-Reperfusion Injury

作者：Johanna T. Kurzhagen, Sanjeev Noel, Kyungho Lee, Mohanraj Sadasivam, Sepideh Gharaie, Aparna Ankireddy, Sul A Lee, Andrea M. Newman-Rivera, Jing Gong, Lois J. Arend, Abdel R.A. Hamad, Sekhar P. Reddy, Hamid Rabb · 发表于：Antioxidants and Redox Signaling · 年份：2023 · DOI：10.1089/ars.2022.0058 · 被引用次数：18 · 研究领域：Renal Transplantation Outcomes and Treatments、Immune Cell Function and Interaction、CRISPR and Genetic Engineering

Aims: 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 Keap1 -knockout (KO) augments Nrf2 antioxidant potential of CD4+ T cells, and that Keap1 -KO CD4+ T cell immunotherapy protects from kidney IRI. Results: CD4+ T cell Keap1 -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. Keap1 -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 in vitro . In vivo , adoptive transfer of Keap1 -KO CD4+ T cells before IRI improved kidney function in T cell-deficient nu/nu mice compared with mice receiving unedited control CD4+ T cells. Keap1 -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. Innovation: 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. Conclusion: CRISPR/Cas9-mediated Keap1 -KO increa...