Aberrant NAD synthetic flux in podocytes under diabetic conditions and effects of indoleamine 2,3-dioxygenase on promoting de novo NAD synthesis

• Published in: Biochemical and biophysical research communications 2023-02, Vol.643, p.61-68
• Main Authors: Zhang, Yuhua, Zhao, Xingchen, Li, Cuili, Yang, Yan, Li, Luan, Chen, Yingwen, Shi, Qingying, Li, Zhilian, Wu, Yanhua, Zhang, Li, Li, Ruizhao, Si, Meijun, Liang, Xinling, Chen, Yuanhan
• Format: Article
• Language: English
• Subjects: Diabetes Mellitus; Diabetic kidney disease; Diabetic Nephropathies; Energy metabolism; Humans; Indoleamine 2,3-dioxygenase; Indoleamine-Pyrrole 2,3,-Dioxygenase - genetics; Indoleamine-Pyrrole 2,3,-Dioxygenase - metabolism; Ligases; NAD - metabolism; Nicotinamide adenine dinucleotide; Podocytes; Podocytes - metabolism
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2022.12.059

Nicotinamide adenine dinucleotide (NAD) is an essential coenzyme in the kidney. The first step in de novo NAD synthesis is regulated by indoleamine 2,3-dioxygenase (IDO), a tryptophan-catabolizing enzyme. Here, we investigated NAD synthetic flux and NAD levels in podocytes under diabetic conditions. We also studied the effects of IDO overexpression on NAD synthetic flux and high glucose (HG)-induced podocyte injury. NAD synthetases in the de novo, Preiss-Handler and salvage pathways were analyzed using in vivo single-nucleus RNA sequencing datasets (GSE131882) of control and diabetic kidney disease (DKD). The mRNA levels of these NAD synthetases were measured in vitro in HG-treated podocytes. The effects of IDO on NAD synthesis were examined by transducing cultured podocytes with an adenovirus encoding IDO, and apoptosis, podocyte markers and mobility were investigated. Cellular transcriptome analysis revealed that control podocytes had relatively low levels of NAD synthetases. In DKD podocytes, de novo NAD synthetase levels were further downregulated. IDO levels were virtually undetectable and did not increase in DKD. In vitro experiments confirmed aberrant de novo NAD synthetic flux and decreased IDO levels in HG-treated podocytes. Overexpression of IDO promoted NAD de novo synthesis, reduced NAD-bypass metabolic enzyme, increased NAD content and recovered podocyte injury markers under diabetic conditions. Taken together, our findings suggest that the de novo NAD synthetic flux is aberrant in DKD, and IDO promotes de novo NAD synthesis and NAD levels, as well as alleviates injury in HG-treated podocytes. •In vivo transcriptome analysis revealed intrinsic low-level NAD synthetase in podocytes at a single cellular level.•The de novo NAD synthetic flux is aberrant under diabetic conditions.•IDO promotes de novo NAD synthesis and NAD levels in HG-treated podocytes.•IDO attenuates HG-induced podocyte injury.