title: Quantitative Differences in the Urinary Proteome of Siblings Discordant for Type 1 Diabetes Include Lysosomal Enzymes.
abstract: Individuals with type 1 diabetes (T1D) often have higher than normal blood glucose levels, causing advanced glycation end product formation and inflammation and increasing the risk of vascular complications years or decades later. To examine the urinary proteome in juveniles with T1D for signatures indicative of inflammatory consequences of hyperglycemia, we profiled the proteome of 40 T1D patients with an average of 6.3 years after disease onset and normal or elevated HbA1C levels, in comparison with a cohort of 41 healthy siblings. Using shotgun proteomics, 1036 proteins were identified, on average, per experiment, and 50 proteins showed significant abundance differences using a Wilcoxon signed-rank test (FDR q-value ≤ 0.05). Thirteen lysosomal proteins were increased in abundance in the T1D versus control cohort. Fifteen proteins with functional roles in vascular permeability and adhesion were quantitatively changed, including CD166 antigen and angiotensin-converting enzyme 2. α-N-Acetyl-galactosaminidase and α-fucosidase 2, two differentially abundant lysosomal enzymes, were detected in western blots with often elevated quantities in the T1D versus control cohort. Increased release of proteins derived from lysosomes and vascular epithelium into urine may result from hyperglycemia-associated inflammation in the kidney vasculature.
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Moo-Jin
authors.initials: MJ
authors.lastname: Suh
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Andrey
authors.initials: A
authors.lastname: Tovchigrechko
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Vishal
authors.initials: V
authors.lastname: Thovarai
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Rolfe
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Manolito G
authors.initials: MG
authors.lastname: Torralba
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Junmin
authors.initials: J
authors.lastname: Wang
authors.affiliation: ‡Pacific Northwest National Laboratory, 902 Battelle Boulevard, P.O. Box 999, Richland, Washington 99352, United States.
authors.forename: Joshua N
authors.initials: JN
authors.lastname: Adkins
authors.affiliation: ‡Pacific Northwest National Laboratory, 902 Battelle Boulevard, P.O. Box 999, Richland, Washington 99352, United States.
authors.forename: Bobbie-Jo M
authors.initials: BJ
authors.lastname: Webb-Robertson
authors.affiliation: §Children's National Medical Center, 111 Michigan Avenue North West, Washington, DC 20010, United States.
authors.forename: Whitney
authors.initials: W
authors.lastname: Osborne
authors.affiliation: §Children's National Medical Center, 111 Michigan Avenue North West, Washington, DC 20010, United States.
authors.forename: Fran R
authors.initials: FR
authors.lastname: Cogen
authors.affiliation: §Children's National Medical Center, 111 Michigan Avenue North West, Washington, DC 20010, United States.
authors.forename: Paul B
authors.initials: PB
authors.lastname: Kaplowitz
authors.affiliation: ‡Pacific Northwest National Laboratory, 902 Battelle Boulevard, P.O. Box 999, Richland, Washington 99352, United States.
authors.forename: Thomas O
authors.initials: TO
authors.lastname: Metz
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Karen E
authors.initials: KE
authors.lastname: Nelson
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Ramana
authors.initials: R
authors.lastname: Madupu
authors.affiliation: †J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, United States.
authors.forename: Rembert
authors.initials: R
authors.lastname: Pieper
mesh_terms: D020410:Activated-Leukocyte Cell Adhesion Molecule / Q000378:metabolism / Q000652:urine; D000293:Adolescent; D000085962:Angiotensin-Converting Enzyme 2; D015153:Blotting, Western; D002648:Child; D002853:Chromatography, Liquid; D015331:Cohort Studies; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000378:metabolism / Q000652:urine*; D004798:Enzymes / Q000378:metabolism / Q000652:urine*; D005260:Female; D006801:Humans; D008247:Lysosomes / Q000201:enzymology / Q000378:metabolism; D008297:Male; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism / Q000652:urine; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D035781:Siblings; D053719:Tandem Mass Spectrometry; D005644:alpha-L-Fucosidase / Q000378:metabolism / Q000652:urine; D048809:alpha-N-Acetylgalactosaminidase / Q000378:metabolism / Q000652:urine
chemical_list: D020410:Activated-Leukocyte Cell Adhesion Molecule; D004798:Enzymes; D020543:Proteome; D048809:alpha-N-Acetylgalactosaminidase; D005644:alpha-L-Fucosidase; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; D000085962:Angiotensin-Converting Enzyme 2
keywords: CD166; Urinary proteome; diabetic vasculature; endothelial barrier function; lysosomal enzyme; protein biomarker; shotgun proteomics; type 1 diabetes; α-fucosidase