High-throughput screening and bioinformatic analysis to ascertain compounds that prevent saturated fatty acid-induced β-cell apoptosis

Seung-Hee Lee; Daniel Cunha; Carlo Piermarocchi; Giovanni Paternostro; Anthony Pinkerton; Laurence Ladriere; Piero Marchetti; Decio L Eizirik; Miriam Cnop; Fred Levine

doi:10.1016/j.bcp.2017.05.007

High-throughput screening and bioinformatic analysis to ascertain compounds that prevent saturated fatty acid-induced β-cell apoptosis

Biochem Pharmacol. 2017 Aug 15:138:140-149. doi: 10.1016/j.bcp.2017.05.007. Epub 2017 May 15.

Authors

Affiliations

¹ Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.
² ULB Center for Diabetes Research, Medical Faculty, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium.
³ Department of Physics and Astronomy, Michigan State University, East Lansing, MI 48824, USA.
⁴ Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
⁵ ULB Center for Diabetes Research, Medical Faculty, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium; Division of Endocrinology, Erasmus Hospital, Université Libre de Bruxelles (ULB), B-1070 Brussels, Belgium.
⁶ Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA. Electronic address: [email protected].

Abstract

Pancreatic β-cell lipotoxicity is a central feature of the pathogenesis of type 2 diabetes. To study the mechanism by which fatty acids cause β-cell death and develop novel approaches to prevent it, a high-throughput screen on the β-cell line INS1 was carried out. The cells were exposed to palmitate to induce cell death and compounds that reversed palmitate-induced cytotoxicity were ascertained. Hits from the screen were analyzed by an increasingly more stringent testing funnel, ending with studies on primary human islets treated with palmitate. MAP4K4 inhibitors, which were not part of the screening libraries but were ascertained by a bioinformatics analysis, and the endocannabinoid anandamide were effective at inhibiting palmitate-induced apoptosis in INS1 cells as well as primary rat and human islets. These targets could serve as the starting point for the development of therapeutics for type 2 diabetes.

Keywords: Beta-cell; Diabetes; Islet; Lipotoxicity.

Publication types

Validation Study

MeSH terms

Animals
Apoptosis / drug effects*
Cell Line
Cells, Cultured
Class I Phosphatidylinositol 3-Kinases
Computational Biology
Enzyme Inhibitors / pharmacology*
Fatty Acids, Nonesterified / adverse effects
Fatty Acids, Nonesterified / antagonists & inhibitors
Female
High-Throughput Screening Assays
Humans
Hypoglycemic Agents / pharmacology*
Insulin-Secreting Cells / cytology
Insulin-Secreting Cells / drug effects*
Insulin-Secreting Cells / metabolism
Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Islets of Langerhans / cytology
Islets of Langerhans / drug effects
Islets of Langerhans / metabolism
Male
Phosphatidylinositol 3-Kinases / genetics
Phosphatidylinositol 3-Kinases / metabolism
Phosphoinositide-3 Kinase Inhibitors*
Protein Kinase Inhibitors / pharmacology*
Protein Serine-Threonine Kinases / antagonists & inhibitors*
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism
RNA Interference
Rats, Wistar
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Small Molecule Libraries
Tissue Culture Techniques

Substances

Enzyme Inhibitors
Fatty Acids, Nonesterified
Hypoglycemic Agents
Intracellular Signaling Peptides and Proteins
Phosphoinositide-3 Kinase Inhibitors
Protein Kinase Inhibitors
Recombinant Proteins
Small Molecule Libraries
MAP4K4 protein, human
Class I Phosphatidylinositol 3-Kinases
PIK3CB protein, human
Protein Serine-Threonine Kinases

Grants and funding

R01 GM122085/GM/NIGMS NIH HHS/United States