Multiparametric Bulk and Single Extracellular Vesicle Pipeline for Identifying Adipose-Specific Signatures in Matched...

Area of Science:

• Endocrinology
• Cell Biology
• Metabolic Disease Research

Background:

• Adipose tissue is an endocrine organ influencing metabolic health.
• Adipose-derived extracellular vesicles (ADEVs) play a role in obesity pathobiology.
• Current methods for isolating and characterizing human ADEVs are insufficient.

Purpose of the Study:

• To develop a robust multiparametric framework for isolating and characterizing human ADEVs.
• To identify specific protein and mRNA signatures of ADEVs in obesity.
• To establish a foundation for an 'adiposity EV signature' in obesity-driven diseases.

Main Methods:

• Developed a multiparametric framework using bulk and single EV characterization, proteomics, and mRNA phenotyping.
• Analyzed EVs from human visceral adipose tissue, adipocyte-conditioned media, and plasma from bariatric surgery patients.
• Utilized bottom-up proteomics, SP-IRIS, and TIRF microscopy for comprehensive analysis.

Main Results:

• Identified unique and shared proteins in adipose tissue-derived EVs (ATEVs) and adipocyte EVs (aEVs).
• Discovered distinct enriched pathways in ATEVs (lipid metabolism, immune modulation) and aEVs (chromatin remodeling, oxidative stress).
• Validated obesity-associated proteins (TMEM120A, SACM1L) and histone variants in ADEVs, and confirmed adiponectin and perilipin in plasma EVs.

Conclusions:

• The developed framework enables tissue and cell-specific characterization of ADEVs.
• Identified specific molecular signatures ('adiposity EV signature') associated with obesity.
• This research provides a foundation for understanding ADEV roles in obesity and related diseases.