A bioluminescent assay for measuring glucose uptake

Area of Science:

• Biochemistry
• Cell Biology
• Assay Development

Background:

• Identifying modulators of glucose uptake is crucial for managing diabetes and developing cancer therapies.
• Existing methods for measuring glucose transport often involve radioactivity or are less user-friendly.
• There is a need for robust, nonradioactive assays to study glucose uptake in various cell types.

Purpose of the Study:

• To develop and validate a novel bioluminescent assay for quantifying cellular glucose uptake.
• To establish an easy-to-use, nonradioactive method for screening glucose transporter inhibitors.
• To assess the assay's utility in measuring insulin sensitivity and its performance across different cell models.

Main Methods:

• The assay utilizes the uptake of 2-deoxyglucose and subsequent enzymatic detection of 2-deoxyglucose-6-phosphate.
• Bioluminescence is measured to quantify accumulated 2-deoxyglucose-6-phosphate, indicating glucose transport.
• The assay was validated using HCT 116 cells, adipocytes, myotubes, and human induced pluripotent stem cell (iPSC)-derived cells.

Main Results:

• The bioluminescent assay demonstrated sensitivity, detecting glucose uptake in as few as 5000 cells with linearity up to 50,000 cells.
• Signal-to-background ratios ranged from 5 to 45, indicating robust detection.
• The assay successfully identified inhibition by known glucose transporter inhibitors and measured insulin-dependent glucose uptake in various cell types, including iPSC-derived cardiomyocytes and skeletal myoblasts.

Conclusions:

• A novel, sensitive, and nonradioactive bioluminescent assay for measuring glucose uptake has been developed.
• This assay is suitable for screening glucose transporter inhibitors and evaluating insulin sensitivity across diverse cellular models.
• The assay provides a valuable tool for diabetes research, cancer therapy development, and stem cell biology.