Profiling of Plasma Metabolites Suggests Altered Mitochondrial Fuel Usage and Remodeling of Sphingolipid Metabolism in Individuals With Type 2 Diabetes and Kidney Disease

Abstract

Introduction: Pathophysiology of diabetic kidney disease (DKD) is incompletely understood. We aim to elucidate metabolic abnormalities associated with DKD in type 2 diabetes mellitus (T2DM) by targeted plasma metabolomics.

Methods: A total of 126 T2DM participants with early DKD (urinary albumin-to-creatinine ratio [ACR] 30-299 mg/g and eGFR ≥ 60 ml/min/1.73 m²), 154 overt DKD (ACR ≥ 300 mg/g or eGFR < 60 ml/min/1.73 m²), and 129 non-DKD T2DM controls (ACR < 30 mg/g and eGFR ≥ 60 ml/min/1.73 m²) were included in discovery study. Findings were subsequently validated in 149 T2DM with macroalbuminuria (ACR ≥ 300 mg/g) and 149 matched non-DKD T2DM controls. Plasma amino acid, acylcarnitine, Krebs cycle organic acid, and sphingolipids/ceramide levels were quantified by liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.

Results: Of 123 metabolites included in the data analysis, 24 differed significantly between DKD and controls in the same direction in both discovery and validation subpopulations. A number of short acylcarnitines including their dicarboxylic derivatives (C2-C6) were elevated in DKD, suggesting abnormalities in fatty acids and amino acids metabolic pathways. Five phosphatidylcholines were lower whereas 4 metabolites in the sphingomyelin-ceramide subfamily were higher in DKD. Principal component regression revealed that long-chain ceramides were independently associated with ACR but not eGFR. Conversely, essential amino acids catabolism and short dicarboxylacylcarnitine accumulation were associated with eGFR but not ACR.

Discussion: DKD is associated with altered fuel substrate use and remodeling of sphingolipid metabolism in T2DM with DKD. Associations of albuminuria and impaired filtration function with distinct metabolomic signatures suggest different pathophysiology underlying these 2 manifestations of DKD.

Keywords: diabetic kidney disease; energetic fuel substrate; metabolomics; pathophysiology; type 2 diabetes mellitus.

Figure 1

Plasma metabolites that differed significantly between diabetic kidney disease (DKD) patients and non-DKD controls in the same direction in both discovery and validation subpopulations (highlighted in red) and the potential metabolic pathways associcated with DKD. AC, acylcarnitine; CPT-1, carnitine palmitoyltransferase I; ER, endoplasmic reticulum; FFA, free fatty acid; LCDA, long-chain dicarboxylic acid; NF-κB, nuclear factor−κB; PDC, pyruvate dehydrogenase complex; SCDA, short-chain diacids; SM, sphingomyelin; TCA, tricarboxylic acid cycle.