Diabetic nephropathy (DN) is a major cause of end-stage renal disease and has limited therapeutic options to prevent its progression. To identify novel therapeutic strategies, protective factors for DN were studied using proteomics on glomeruli from 50-year medalists (individuals with extreme duration, ≥50 years, of diabetes in Joslin Medalist Study) without DN and those with histologic signs of DN. Many enzymes in the glycolytic, sorbitol, methylglyoxal and mitochondrial pathways were elevated in individuals without DN. We also identified that pyruvate kinase M2 (PKM2) expression and activity were upregulated. We then looked into the mechanisms and further validated the findings in vivo (Podocyte-specific Pkm2 KO mice model) and in vitro (cultured podocytes).