Postischemic inactivation of HIF prolyl hydroxylases in endothelium promotes maladaptive kidney repair by inducing glycolysis
Ratnakar Tiwari, … , Navdeep S. Chandel, Pinelopi P. Kapitsinou
Ratnakar Tiwari, … , Navdeep S. Chandel, Pinelopi P. Kapitsinou
Published December 2, 2024
Citation Information: J Clin Invest. 2025;135(3):e176207. https://doi.org/10.1172/JCI176207.
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Research Article Metabolism Nephrology

Postischemic inactivation of HIF prolyl hydroxylases in endothelium promotes maladaptive kidney repair by inducing glycolysis

Abstract

Ischemic acute kidney injury (AKI) is common in hospitalized patients and increases the risk for chronic kidney disease (CKD). Impaired endothelial cell (EC) functions are thought to contribute in AKI to CKD transition, but the underlying mechanisms remain unclear. Here, we identify a critical role for endothelial oxygen sensing prolyl hydroxylase domain (PHD) enzymes 1–3 in regulating postischemic kidney repair. In renal endothelium, we observed compartment-specific differences in the expression of the 3 PHD isoforms in both mice and humans. Postischemic concurrent inactivation of endothelial PHD1, PHD2, and PHD3 but not PHD2 alone promoted maladaptive kidney repair characterized by exacerbated tissue injury, fibrosis, and inflammation. scRNA-Seq analysis of the postischemic endothelial PHD1, PHD2, and PHD3-deficient (PHDTiEC) kidney revealed an endothelial hypoxia and glycolysis-related gene signature, also observed in human kidneys with severe AKI. This metabolic program was coupled to upregulation of the SLC16A3 gene encoding the lactate exporter monocarboxylate transporter 4 (MCT4). Strikingly, treatment with the MCT4 inhibitor syrosingopine restored adaptive kidney repair in PHDTiEC mice. Mechanistically, MCT4 inhibition suppressed proinflammatory EC activation, reducing monocyte-EC interaction. Our findings suggest avenues for halting AKI to CKD transition based on selectively targeting the endothelial hypoxia-driven glycolysis/MCT4 axis.

Authors

Ratnakar Tiwari, Rajni Sharma, Ganeshkumar Rajendran, Gabriella S. Borkowski, Si Young An, Michael Schonfeld, James O’Sullivan, Matthew J. Schipma, Yalu Zhou, Guillaume Courbon, Benjamin R. Thomson, Valentin David, Susan E. Quaggin, Edward B. Thorp, Navdeep S. Chandel, Pinelopi P. Kapitsinou

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Figure 9

Syrosingopine or MCT4 knockdown suppresses the expression of EC adhesion molecules in HPAECs activated by hypoxia/reoxygenation and IL-1β.

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Syrosingopine or MCT4 knockdown suppresses the expression of EC adhesion...
(A) Experimental schematic for HPAECs subjected to 0.5% O2 for 18 hours in the presence of syrosingopine (5 μM) or MCT4 siRNA followed by reoxygenation for 8 hours in the presence of IL-1β (1 ng/ml). (B) mRNA levels of VCAM1 and ICAM1 in syrosingopine- vs vehicle-treated HPAECs, that were activated by hypoxia/reoxygenation and IL-1β. (C) THP1 monocyte adhesion to inflamed ECs. THP1 monocyte cells, labeled with green CMFDA dye, were introduced on a monolayer of HPAECs that had been subjected to the indicated experimental conditions. Following a 90-minute incubation period, floating cells were washed away and adhered THP1 cells were visualized using a fluorescent microscope and subsequently quantified. Representative images of fluorescent THP1 cells attached to ECs in different experimental groups are presented. Scale bar: 200 μm. (D) mRNA expression of VCAM1 and ICAM1 in HPAECs transfected with control or MCT4 siRNA and subjected to hypoxia/reoxygenation and IL-1β. (E) THP1 monocyte adhesion to inflamed ECs under the same experimental conditions as in D. Scale bar: 200 μm. Data are represented as mean ± SEM. Statistics were determined by 1-way ANOVA with Šidák’s correction for multiple comparisons. n = 3–4. *P < 0.05; **P <0.01; ***P <0.001; ****P < 0.0001. Nx, normoxia; Hx, hypoxia/reoxygenation; C, negative control siRNA; MCT4si, MCT4siRNA.