Insulin Promoter-Driven Gaussia Luciferase-Based Insulin Secretion Biosensor Assay for Discovery of β-Cell Glucose-Sensing Pathways

Michael A. Kalwat, Chonlarat Wichaidit, Alejandra Y. Nava Garcia, Melissa K. McCoy, Kathleen McGlynn, In Hyun Hwang, John B. Macmillan, Bruce A. Posner, Melanie H. Cobb

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

High throughput screening of insulin secretion is intractable with current methods. We developed a secreted insulin-luciferase system (Ins-GLuc) in β cells that is rapid, inexpensive, and amenable to 96- and 384-well formats. We treated stable Ins-GLuc-expressing MIN6 cells overnight with 6298 marine natural product fractions. The cells were then washed to remove media and chemicals, followed by stimulation with glucose in the diazoxide paradigm. These conditions allowed the discovery of many insulin secretion suppressors and potentiators. The mechanisms of action of these natural products must be long-lasting given the continuance of secretory phenotypes in the absence of chemical treatment. We anticipate that these natural products and their target pathways will lead to a greater understanding of glucose-stimulated insulin secretion.

Original languageEnglish (US)
Pages (from-to)1208-1212
Number of pages5
JournalACS Sensors
Volume1
Issue number10
DOIs
StatePublished - 2016

Keywords

  • Gaussia
  • high-throughput screening
  • insulin
  • marine natural products
  • pancreatic islet beta cell

ASJC Scopus subject areas

  • Bioengineering
  • Instrumentation
  • Process Chemistry and Technology
  • Fluid Flow and Transfer Processes

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