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bioRxiv · 10.1101/2021.06.11.448037

Alpha-ketoglutarate augments prolyl hydroxylase-2 mediated inactivation of phosphorylated-Akt to inhibit induced-thrombosis and inflammation

Shrimali, N. M.·Agarwal, S.·Kaur, S.·Bhattacharya, S.·Bhattacharyya, S.·Prchal, J. T.·Guchhait, P.
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Abstract

Phosphorylation of Akt (pAkt) regulates multiple physiological and pathological processes including thrombosis and inflammation. In an approach to inhibit the pathological signalling of pAkt by prolyl-hydroxylase-2 (PHD2) we employed -ketoglutarate (KG), a cofactor of PHD2. Octyl-KG supplementation to platelets promoted PHD2 activity through elevated intracellular KG:succinate ratio and reduced aggregation in vitro by suppressing pAkt1(Thr308). Augmented PHD2 activity was confirmed by increased hydroxylated-proline alongside enhanced binding of PHD2 to pAkt in KG-treated platelets. Contrastingly, inhibitors of PHD2 significantly increased pAkt1 in platelets. Octyl-KG followed similar mechanism in monocytes to inhibit cytokine secretion in vitro. Our data also describe a suppressed pAkt1 and reduced activation of platelet and leukocyte obtained from mice supplemented with dietary-KG, unaccompanied by alteration in their counts. Dietary-KG significantly reduced clot formation and leukocyte accumulation in various organs including lung of mice treated with thrombosis-inducing agent carrageenan. Importantly, we observed a significant rescue effect of dietary-KG on inflamed lung of SARS-CoV-2 infected hamsters. KG significantly reduced leukocyte accumulation, clot formation and viral load alongside downmodulation of pAkt in lung of the infected animals. Therefore, our study suggests a safe implementation of dietary-KG in prevention of Akt-driven anomalies including thrombosis and inflammation, highlighting a better pulmonary management in COVID-19.

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Shrimali, N. M., Agarwal, S., Kaur, S., Bhattacharya, S., Bhattacharyya, S., Prchal, J. T., Guchhait, P.. 2021-06-12. Alpha-ketoglutarate augments prolyl hydroxylase-2 mediated inactivation of phosphorylated-Akt to inhibit induced-thrombosis and inflammation. https://doi.org/10.1101/2021.06.11.448037

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