Proteomic analysis identifies dysregulated proteins in albuminuria: a South African pilot study

Abstract

Abstract Albuminuria may precede decreases in glomerular filtration rate (GFR) and both tests are insensitive predictors of early stages of kidney disease. Our aim was to characterise the urinary proteome in black African individuals with albuminuria and well-preserved GFR from South Africa. A case-controlled study that compared urinary proteomes of 52 normoalbuminuric (urine albumin: creatinine ratio (uACR) <3 mg/mmol) and 56 albuminuric (uACR ≥ 3 mg/mmol) adults of Black African ethnicity. Urine proteins were precipitated, reduced, alkylated, digested, and analysed using an Evosep One LC coupled to a Sciex 5600 Triple-TOF in data-independent acquisition mode. Data were searched on Spectronaut TM 15. Differentially abundant proteins (DAPs) were filtered ≥ 2.25-fold change and false discovery rate ≤ 1%. Receiver operating characteristic curves were used to assess the discriminating ability of proteins of interest. Pathway analysis was performed using Enrichr software. The albuminuric group had a higher uACR (7.9 vs 0.55 mg/mmol, p <0.001). The median eGFR (mL/min/1.73m 2 ) showed no difference between the groups (111 vs 114, p =0.707). We identified 80 DAPs in the albuminuria group compared to normoalbuminuria, of which 59 proteins increased while 21 proteins decreased in abundance. We found 12 urinary proteins with AUC > 0.8, and p -value <0.001 in the multivariate analysis. Furthermore, an 80-protein model was developed that showed a high AUC >0.907 and a predictive accuracy of 91.3% between the two groups. Pathway analysis associated with DAPs were involved in insulin growth factor (IGF) functions, innate immunity, platelet degranulation, and extracellular matrix organization. In albuminuric individuals with well-preserved eGFR, pathways involved in preventing the release and uptake of IGF by insulin growth factor binding protein were significantly enriched. These proteins are indicative of a homeostatic imbalance in a variety of cellular processes underlying renal dysfunction and are implicated in chronic kidney disease.