*Purpose: T cells play an important role in the pathophysiology of kidney ischemia reperfusion injury (IRI). Metabolic programming of T cells, a rapidly evolving field, controls T cell function, but little is known in kidney IRI. We hypothesized that kidney T cell metabolism is dysregulated during IRI and modulation can improve outcome.

*Methods: C57BL/6J mice underwent bilateral IRI with 30 min ischemia, and kidneys and spleens were harvested at multiple early time points including during ischemia. Human non-ischemic and ischemic kidney tissues were procured from normal portions of renal cell carcinoma nephrectomy kidneys. T cells were isolated and studied with a spectral flow cytometry-based immunometabolic assay that allows the interrogation of T cell metabolic programs at the single cell level. The data was then evaluated by unbiased computational analyses with a machine learning algorithm. The glutamine antagonist JHU083, which targets T cell metabolism, was administered intraperitoneally and effects on IRI were measured.

*Results: Unbiased multi-dimensional analyses revealed a distinct T cell population having low expression of phospho-S6 ribosomal protein (pS6), a surrogate marker of mTOR activity, and mitochondrial voltage dependent anion channel 1 in post-ischemic kidneys. An epigenetic marker H3K27Me3 expression, regulated by multiple metabolites in TCA cycle, drove the segregation of ischemic kidney T cells from those of non-ischemic kidneys in both humans and mice. Splenic T cells from post-AKI mice showed higher expression of GLUT1, hexokinase II (HKII), and CPT1a, indicating an upregulation of glycolysis and fatty acid oxidation machinery. Blocking glutamine utilization by JHU083 treatment ameliorated renal injury at 24h (plasma creatinine, vehicle control vs JHU083, 1.7±0.8 vs 1.0±0.5 mg/dL, P=0.03), and the proportion of intact tubules in renal medulla were higher in post-ischemic kidneys of JHU083 treated mice (27±11 vs 43±17%, P=0.03). Expression of pS6 (normalized MFI 0.38±0.07 vs 0.47±0.06, P<0.01) and HKII (0.31±0.04 vs 0.41±0.05, P<0.01) were enhanced in kidney T cells from JHU083-treated mice, compared to vehicle-treated mice. CD4⁺ T cells (CD44, 67±5 vs 56±6%, P<0.01; Ki67, 59±9 vs 51±6%, P=0.03) and CD8⁺ T cells (CD44, 61±11 vs 41±11%, P<0.01; CD69, 25±7 vs 18±4%, P=0.02; Ki67, 61±13 vs 48±12%, P=0.04) from post-ischemic kidneys of the JHU083-treated mice showed reduced activation and proliferation, whereas they increased in double-negative T cells (CD44, 94±2 vs 96±1%, P=0.04; CD69, 61±8 vs 72±7%, P<0.01; Ki67, 79±9% vs 91±3%, P<0.01).

*Conclusions: Kidney IRI induces metabolic reprogramming in human and murine kidney T cells. Reconstitution of T cells metabolism by blocking glutamine utilization with JHU083 is a promising novel approach to improve quantity and quality of deceased donor organs for transplant.

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