Back to Search
Start Over
A Class of Diacylglycerol Acyltransferase 1 Inhibitors Identified by a Combination of Phenotypic High-throughput Screening, Genomics, and Genetics
- Source :
- Scipedia Open Access, Scipedia SL, EBioMedicine, EBioMedicine, Vol 8, Iss C, Pp 49-59 (2016)
- Publication Year :
- 2016
- Publisher :
- Elsevier BV, 2016.
-
Abstract
- Excess lipid storage is an epidemic problem in human populations. Thus, the identification of small molecules to treat or prevent lipid storage-related metabolic complications is of great interest. Here we screened > 320.000 compounds for their ability to prevent a cellular lipid accumulation phenotype. We used fly cells because the multifarious tools available for this organism should facilitate unraveling the mechanism-of-action of active small molecules. Of the several hundred lipid storage inhibitors identified in the primary screen we concentrated on three structurally diverse and potent compound classes active in cells of multiple species (including human) and negligible cytotoxicity. Together with Drosophila in vivo epistasis experiments, RNA-Seq expression profiles suggested that the target of one of the small molecules was diacylglycerol acyltransferase 1 (DGAT1), a key enzyme in the production of triacylglycerols and prominent human drug target. We confirmed this prediction by biochemical and enzymatic activity tests.<br />Highlights • We identified > 600 potent small molecule inhibitors of cellular lipid storage deposition. • RNA-Seq expression profiling discriminated the activity of three lead scaffolds and guided subsequent functional studies. • We discovered a class of DGAT1 inhibitors, which is active in fly and mammalian cell lines as well as whole flies. Obesity and other lipid storage associated diseases are a growing health threat of human populations. In an undirected phenotypic screen, we identified pharmacologically active small molecules that reduce or enhance lipid storage. Our work focuses on three lead structures that prevent lipid storage in diverse cellular systems including cells from a diabetes patient. In order to elucidate the compound mechanisms-of-action and cellular targets, we used a combination of RNA-Seq transcriptional profiling and diverse functional assays. Our results strongly suggest that one of our lead structures represents a class of inhibitors targeting the key lipogenic enzyme diacylglycerol acyltransferase 1.
- Subjects :
- Male
0301 basic medicine
High-throughput screening
lcsh:Medicine
Genomics
Biology
General Biochemistry, Genetics and Molecular Biology
Cell Line
Small Molecule Libraries
Mice
Structure-Activity Relationship
03 medical and health sciences
Medicine, General & Internal
In vivo
Chlorocebus aethiops
Animals
Humans
Structure–activity relationship
Pyrroles
Diacylglycerol O-Acyltransferase
Enzyme Inhibitors
Genetics
chemistry.chemical_classification
lcsh:R5-920
Sequence Analysis, RNA
Fatty Acids
lcsh:R
Cell Differentiation
Epistasis, Genetic
General Medicine
Phenotype
Small molecule
3. Good health
030104 developmental biology
Enzyme
Biochemistry
chemistry
COS Cells
Epistasis
Drosophila
Female
lipids (amino acids, peptides, and proteins)
Lipid Peroxidation
lcsh:Medicine (General)
Research Paper
Subjects
Details
- ISSN :
- 23523964
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- EBioMedicine
- Accession number :
- edsair.doi.dedup.....f3adc3091713c88fcd187f80aef16eb8