Introduction and Objective: Type 1 diabetes (T1D) is an autoimmune disease that causes loss of β-cell mass and insulin secretion. Early Stage 3 T1D includes β-cell dysfunction and activation of intrinsic stress pathways before clinical diagnosis, making this ‘asymptomatic’ period critical for prediction and prevention. Circulating plasma extracellular vesicles (EVs) are promising non-invasive biomarkers for disease identification and prognosis. Thus, the objective is to determine an early EV-based prognostic indicator of declining β-cell health in T1D.Methods: Adult and pediatric human blood specimens were collected from three groups: 1) Early (E)T1D (<1 year from diagnosis, n=10); 2) Late (L)T1D (>1 year from diagnosis, n=9); 3) age-matched healthy controls (NC, n=7). Blood plasma-derived EVs were isolated using size-exclusion chromatography and differential ultracentrifugation. EVs were characterized through Nanoparticle Tracking Analysis (NTA), western blotting, assessment of EVs on β-cell function, and proteomic analysis using tandem mass spectrometry.Results: NTA revealed a 2-fold increase in plasma EVs from ET1D and LT1D groups compared to NC (p<0.05). Human islets exposed to LT1D EV but not ET1D and NC EVs, showed impaired glucose-stimulated insulin secretion (GSIS) (2 X 109 particles/day for 48 h). Proteomic analysis identified 1,229 enriched EV proteins, and differential expression analysis (log2(fold change) ≥ 1.5) highlighted potential biomarkers for ET1D, including VWF, LBP, and GP1BA (p<0.05). KEGG pathway analysis revealed enriched proteins related to complement and coagulation, extracellular matrix, antigen presentation, and immune system pathways in ET1D and LT1D EVs compared to NC EVs. VWF expression was confirmed by western blotting in both ET1D and LT1D EVs but was absent in NC EVs.Conclusion: Our study characterizes circulating EVs from three patient groups, identifying VWF as a potential biomarker for early Stage 3 T1D.
R. Leon-Gutierrez: None. G. Ariyaratne: None. Z. Liang: None. A. Hoff: None. A. Roy: None. M. Tahawi: None. A. Matveyenko: None. A.M. Egan: None. A. Vella: Research Support; Novo Nordisk A/S, Dexcom, Inc. Advisory Panel; Boehringer-Ingelheim, Rezolute. A. Creo: Advisory Panel; Sanofi. N. Javeed: None.
NIH (DK129208-01); JDRF (2-SRA-2022-1272-S-B)
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