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Cell Metab DOI:10.1016/j.cmet.2014.12.010

High-throughput luminescent reporter of insulin secretion for discovering regulators of pancreatic Beta-cell function.

Publication TypeJournal Article
Year of Publication2015
AuthorsBurns, SM, Vetere, A, Walpita, D, Dančík, V, Khodier, C, Perez, J, Clemons, PA, Wagner, BK, Altshuler, D
JournalCell Metab
Date Published2015 Jan 06
KeywordsCells, Cultured, Cytokines, Enzyme-Linked Immunosorbent Assay, Fatty Acids, Genes, Reporter, Glucose, High-Throughput Screening Assays, Humans, Insulin, Insulin Secretion, Insulin-Secreting Cells, Luciferases, Recombinant Fusion Proteins, Thapsigargin

Defects in insulin secretion play a central role in the pathogenesis of type 2 diabetes, yet the mechanisms driving beta-cell dysfunction remain poorly understood, and therapies to preserve glucose-dependent insulin release are inadequate. We report a luminescent insulin secretion assay that enables large-scale investigations of beta-cell function, created by inserting Gaussia luciferase into the C-peptide portion of proinsulin. Beta-cell lines expressing this construct cosecrete luciferase and insulin in close correlation, under both standard conditions or when stressed by cytokines, fatty acids, or ER toxins. We adapted the reporter for high-throughput assays and performed a 1,600-compound pilot screen, which identified several classes of drugs inhibiting secretion, as well as glucose-potentiated secretagogues that were confirmed to have activity in primary human islets. Requiring 40-fold less time and expense than the traditional ELISA, this assay may accelerate the identification of pathways governing insulin secretion and compounds that safely augment beta-cell function in diabetes.


Alternate JournalCell Metab.
PubMed ID25565210