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Abstract: SA-PO269

Plasma and Urine Degradome Integration Reveals Sexual Dimorphic Prognostic Biomarkers for ESKD in Type 1 Diabetes (T1D)

Session Information

Category: Diabetic Kidney Disease

  • 701 Diabetic Kidney Disease: Basic

Authors

  • Merchant, Michael, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Arteel, Gavin E., University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Wilkey, Daniel Wade, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Adams, Lauren T., University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Miller, Rachel G., University of Pittsburgh School of Public Health, Pittsburgh, Pennsylvania, United States
  • Benos, Panayiotis, University of Florida Health, Gainesville, Florida, United States
  • Costacou, Tina, University of Pittsburgh School of Public Health, Pittsburgh, Pennsylvania, United States
Background

DKD is a highly prevalent, severe microvascular T1D complication associated with altered expression/turnover of matrisomal proteins. Proteomics has identified several candidate surrogate biomarkers for DKD progression, but most studies did not consider urine/plasma proteome integration or sexual dimorphic responses. We hypothesized integration of plasma/urine low molecular weight proteome (degradome) could reveal sex-dependent differences associated with T1D ESKD incidence.

Methods

Paired urine & plasma samples for eight age/sex matched ESKD incident cases and non-ESKD controlswere randomly selected from the Pittsburgh Epidemiology of Diabetes Complications (EDC) T1D study biorepository. Samples were shipped to the University of Louisville and processed for degradome analysis using high resolution mass spectrometry in a data dependent analysis (DDA) fashion. De novo spectral assignments used PeaksX software. Uni-/multi-variate analysis compared feature abundances (p<0.05 for significance). MatrisomeDb and TopFinder assigned matrisome composition and imputed likely protease activity. Comparisons were made using XIC areas for (1) individual peptides, (2) summed related, overlapping peptide sequences (degradome domains) and (3) peptides compiled for parent proteins. Exploratory confirmation studies of parent protein abundance were conducted by ELISA.

Results

More than 22,000 peptides, corresponding to 955 parent proteins were observed. 1,792 plasma and 1,041 peptides were observed in more than 50% of the samples with many unique to plasma (11) or urine (69). 191 matrisomal proteins were identified including known T1D DKD markers (COL1A1, BGN, AGT, TNFSF12, and EGF). Significantly enriched imputed proteases included MMP12, MMP7, MMP3, and MMP14. Fragments of fifteen proteins including PGRMC1, ORM1, A1BG, AGT, and APOA2 distinguished all cases from controls. Sexual dimorphic effects were observed for SerpinD1, MGP, AGT and EGF. EGF ELISA confirmed urine EGF degradome findings.

Conclusion

T1D DKD is associated with plasma and urinary degradomic changes observable ~12 yrs before ESKD onset. These differences may provide mechanistic insights into DKD progression and lead to novel surrogate diagnostic and target engagement biomarkers for ESKD intervention.

Funding

  • NIDDK Support