Discovering metabolic disease gene interactions by correlated effects on cellular morphology

Autor: Josep-Maria Mercader, Jason Flannick, John G. Doench, M.F. Michelle Sim, J. Michael Edwardson, Kate Hartland, Sara A Georges, Dermot F. Reilly, Maria Kost-Alimova, Anna I. Podgornaia, Andrea Bejar, Vilas Wagh, Amit R. Majithia, Evan D. Rosen, Justin J. Rochford, Marija Tadin-Strapps, Yang Jiao, Jesse M. Engreitz, Xiaolan Zhang, Robert H. Rice, Mesbah Uddin Talukder, Umer Ahmed
Přispěvatelé: Apollo - University of Cambridge Repository
Rok vydání: 2019
Předmět:
0301 basic medicine
Lipodystrophy
Gene discovery
Physiology
Genetic screen
0302 clinical medicine
GTP-Binding Protein gamma Subunits
CEBPA
Adipocytes
2.1 Biological and endogenous factors
Gene Regulatory Networks
Protein Interaction Maps
Aetiology
Cells
Cultured
Cultured
Adipogenesis
Diabetes
Functional genomics
Type 2 diabetes
Phenotype
Metabolic syndrome
High content imaging
Original Article
Biotechnology
lcsh:Internal medicine
Perilipin-1
Cells
030209 endocrinology & metabolism
Computational biology
Biology
03 medical and health sciences
Insulin resistance
Rare Diseases
medicine
Diabetes Mellitus
Genetics
Humans
Obesity
lcsh:RC31-1245
Molecular Biology
Gene
Metabolic and endocrine
Nutrition
Human Genome
Cell Biology
medicine.disease
030104 developmental biology
HEK293 Cells
CCAAT-Enhancer-Binding Proteins
Biochemistry and Cell Biology
Transcriptome
Acyltransferases
Zdroj: Molecular Metabolism
Molecular Metabolism, Vol 24, Iss, Pp 108-119 (2019)
DOI: 10.17863/cam.38946
Popis: Objective Impaired expansion of peripheral fat contributes to the pathogenesis of insulin resistance and Type 2 Diabetes (T2D). We aimed to identify novel disease–gene interactions during adipocyte differentiation. Methods Genes in disease-associated loci for T2D, adiposity and insulin resistance were ranked according to expression in human adipocytes. The top 125 genes were ablated in human pre-adipocytes via CRISPR/CAS9 and the resulting cellular phenotypes quantified during adipocyte differentiation with high-content microscopy and automated image analysis. Morphometric measurements were extracted from all images and used to construct morphologic profiles for each gene. Results Over 107 morphometric measurements were obtained. Clustering of the morphologic profiles accross all genes revealed a group of 14 genes characterized by decreased lipid accumulation, and enriched for known lipodystrophy genes. For two lipodystrophy genes, BSCL2 and AGPAT2, sub-clusters with PLIN1 and CEBPA identifed by morphological similarity were validated by independent experiments as novel protein–protein and gene regulatory interactions. Conclusions A morphometric approach in adipocytes can resolve multiple cellular mechanisms for metabolic disease loci; this approach enables mechanistic interrogation of the hundreds of metabolic disease loci whose function still remains unknown.
Highlights • Loss-of-function genetic screen in human adipocytes for 125 genes selected from metabolic disease-associated loci. • Genetic screen read out by cellular morphometry— 77,000 images taken with 400 morphological features extracted per image. • Pairwise mechanistic interactions between genes identified by correlations of cellular morphometry—two interactions validated. • Novel interaction between BSCL2 and PLIN1 from biophysical association of proteins at lipid droplet surface. • Novel interaction between CEBPA and AGPAT2 from CEBPA dependent transcription of AGPAT2.
Databáze: OpenAIRE
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