A Real Time Quaking Induced Conversion (RT-QuIC) Assay for Detection of Misfolded Insulin Protein

Area of Science:

• Biochemistry
• Molecular Biology
• Metabolic Disease Research

Background:

• Protein misfolding is implicated in aging and disease, but its role in metabolic dysfunction remains unclear.
• Insulin, essential for glucose regulation, can misfold, potentially contributing to metabolic disorders.

Purpose of the Study:

• To develop and validate a Real-time Quaking-Induced Conversion (RT-QuIC) assay for detecting misfolded insulin.
• To explore the potential of this assay in studying insulin aggregation in metabolic disease models.

Main Methods:

• Adaptation of the RT-QuIC assay, originally for prions, to detect misfolded insulin.
• Validation using recombinant insulin and clinical insulin aggregates.
• Application of the assay to tissue samples from a mouse model of metabolic disease.

Main Results:

• Successful adaptation and validation of the RT-QuIC assay for misfolded insulin detection.
• Characterization of insulin aggregates from clinical devices revealed structural differences.
• Demonstrated proof-of-concept for using the RT-QuIC assay in a metabolic disease mouse model.

Conclusions:

• The developed RT-QuIC assay provides a novel tool for studying insulin aggregation.
• This assay has potential for early detection and understanding of metabolic disorders linked to aging and disease.
• Further research may uncover new therapeutic targets for metabolic dysfunction.