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Abstract: FR-PO1130

Quantitative Mass Spectrometry Aided Urine Biomarker Analysis in Nephrotic Syndrome

Session Information

Category: Glomerular Diseases

  • 1203 Glomerular Diseases: Clinical, Outcomes, and Trials

Authors

  • Cummins, Timothy, University of Louisville Kidney Disease Program, Louisville, Kentucky, United States
  • Wilkey, Daniel Wade, University of Louisville Kidney Disease Program, Louisville, Kentucky, United States
  • Barati, Michelle T., University of Louisville Kidney Disease Program, Louisville, Kentucky, United States
  • Caster, Dawn J., University of Louisville Kidney Disease Program, Louisville, Kentucky, United States
  • Benz, Frederick W., University of Louisville, Louisville, Kentucky, United States
  • Merchant, Michael, University of Louisville Kidney Disease Program, Louisville, Kentucky, United States
  • Klein, Jon B., University of Louisville Kidney Disease Program, Louisville, Kentucky, United States
Background

Accurate and precise diagnosis of chronic renal diseases can be aided using biomarkers. Methodological advancements in mass spectrometry and bioinformatics have improved the ability to identify and quantitate protein biomarkers induced in disease. Proteins and peptides that are present in patient urine are indicative of changes in homeostasis of the kidney and other organs. Identification and quantitation of these components of the urine proteome will be useful in determining effective new biomarkers of kidney disease. Loss of kidney function corresponds with increased leakage of protein and peptide components into the urine. Profiling and quantitation of urinary markers of renal disease progression is important towards a better understanding and characterization of distinct glomerular diseases.

Methods

In this work we present a mass spectrometry workflow utilizing multiplex tandem mass tag (TMT) quantitation of whole urine proteins from patients with nephrotic syndrome and in remission.

Results

Combined 2D-LC-MS/MS-TMT analysis yielded ~800 protein identifications, ~480 were quantitated. Over 200 proteins were upregulated in disease and 50 were downregulated.

Conclusion

Our current study includes urine TMT analysis from 3 patients with nephrotic syndrome and post-treatment follow-up upon remission. The sample set, though small, is ideal for looking at effects of normalization on intra and inter-patient protein quantitation as we were also able to see a difference in the effects of race and sex in one sample that clearly clusters differently from the other two patients. Clinical data, specifically urinary creatinine, was used to normalize TMT values. Employing these methods on larger patient cohorts will help in uncovering novel biomarkers of nephrotic syndrom.

Funding

  • NIDDK Support