iTRAQ-based quantitative protein expression profiling and MRM verification of markers in type 2 diabetes

Area of Science:

• Biochemistry
• Molecular Biology
• Metabolomics

Background:

• Type 2 diabetes mellitus (T2DM) pathogenesis is complex and molecularly heterogeneous.
• Current diagnostics (blood glucose) do not predict disease progression or complications like cardiovascular disease.
• Predicting T2DM complications remains a significant clinical challenge.

Purpose of the Study:

• To identify and validate circulating serum biomarkers for Type 2 diabetes mellitus.
• To explore potential clinical utility in patient stratification and therapeutic development.
• To investigate molecular pathways associated with T2DM pathogenesis.

Main Methods:

• Quantitative proteomic analysis using isobaric tagging for relative and absolute quantification (iTRAQ).
• Validation of candidate protein markers via multiple reaction-monitoring mass spectrometry (MRM-MS), Western blot, and quantitative PCR (QPCR).
• Functional pathway analysis to understand biological processes affected in T2DM.

Main Results:

• Differential protein expression identified between T2DM patients and non-diabetic controls.
• Five protein markers were validated across multiple methodologies.
• Pathway analysis revealed disruptions in lipid metabolism, small molecule metabolism, glucose homeostasis, and blood coagulation.

Conclusions:

• The identified protein biomarkers may aid in subset stratification of T2DM patients.
• These biomarkers hold potential for developing novel therapeutics targeting specific T2DM pathologies.
• Further research can leverage these findings for improved T2DM management and personalized medicine.