Protein N-Glycans in Healthy and Sclerotic Glomeruli in Diabetic Kidney Disease

Veličković, Dušan; Shapiro, John P.; Parikh, Samir V.; Rovin, Brad; Toto, Robert D.; Vazquez, Miguel A.; Poggio, Emilio D.; O'Toole, John F.; Sedor, John R.; Alexandrov, Theodore; Jain, Sanjay; Bitzer, Markus; Hodgin, Jeffrey; Veličković, Marija; Sharma, Kumar; Anderton, Christopher R.

doi:10.1681/asn.0000000000000393

Protein N-Glycans in Healthy and Sclerotic Glomeruli in Diabetic Kidney Disease

Journal Article · Sun Sep 01 00:00:00 EDT 2024 · Journal of the American Society of Nephrology

DOI:https://doi.org/10.1681/asn.0000000000000393· OSTI ID:2506726

Veličković, Dušan ^[1]; Shapiro, John P. ^[2]; ^[2]; Rovin, Brad ^[2]; ^[3]; ^[3]; Poggio, Emilio D. ^[4]; O'Toole, John F. ^[4]; ^[4]; ^[5]; ^[6]; ^[7]; ^[7]; ^[1]; ^[8]; ^[9]

Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
The Ohio State Univ., Columbus, OH (United States)
Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
Cleveland Clinic, OH (United States)
European Molecular Biology Laboratory (EMBL), Heidelberg (France); BioInnovation Institute, Copenhagen (Denmark). BioStudio
Washington Univ., St. Louis, MO (United States). School of Medicine
Univ. of Michigan, Ann Arbor, MI (United States)
Univ. of Texas at San Antonio, TX (United States)
Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Univ. of Texas at San Antonio, TX (United States)

Diabetes is expected to directly affect renal glycosylation; yet to date, there has not been a comprehensive evaluation of alterations in N-glycan composition in the glomeruli of patients with diabetic kidney disease (DKD). Here, we used untargeted mass spectrometry imaging to identify N-glycan structures in healthy and sclerotic glomeruli in formalin-fixed paraffin-embedded sections from needle biopsies of five patients with DKD and three healthy kidney samples. Regional proteomics was performed on glomeruli from additional biopsies from the same patients to compare the abundances of enzymes involved in glycosylation. Secondary analysis of single-nucleus RNA sequencing (snRNAseq) data were used to inform on transcript levels of glycosylation machinery in different cell types and states. We detected 120 N-glycans, and among them, we identified 12 of these protein post-translated modifications that were significantly increased in glomeruli. All glomeruli-specific N-glycans contained an N-acetyllactosamine epitope. Five N-glycan structures were highly discriminant between sclerotic and healthy glomeruli. Sclerotic glomeruli had an additional set of glycans lacking fucose linked to their core, and they did not show tetra-antennary structures that were common in healthy glomeruli. Orthogonal omics analyses revealed lower protein abundance and lower gene expression involved in synthesizing fucosylated and branched N-glycans in sclerotic podocytes. In snRNAseq and regional proteomics analyses, we observed that genes and/or proteins involved in sialylation and N-acetyllactosamine synthesis were also downregulated in DKD glomeruli, but this alteration remained undetectable by our spatial N-glycomics assay. Integrative spatial glycomics, proteomics, and transcriptomics revealed protein N-glycosylation characteristic of sclerotic glomeruli in DKD.

View Accepted Manuscript (DOE)

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE

Contributing Organization:: Kidney Precision Medicine Project

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 2506726

Report Number(s):: PNNL-SA--202058

Journal Information:: Journal of the American Society of Nephrology, Journal Name: Journal of the American Society of Nephrology Journal Issue: 9 Vol. 35; ISSN 1533-3450

Publisher:: American Society of Nephrology (ASN)Copyright Statement

Country of Publication:: United States

Language:: English

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