Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of rat insulinoma (INS-1 derived) cells grown in lipid culture

ABSTRACT: This experiment was designed to study the effects of lipid culture on INS-1 derived glucose-responsive 832/13 cells. After 48-hr culture with lipids, a dramatic decrease in glucose-responsiveness is observed. In order to detect some earlier changes, RNA was also collected at earlier time points (12 hr and 24 hr). To compensate for any changes associated with cell density, control cells (1%BSA culture) were included for each time point. RNA was prepared from INS-1 cells in Dr. Chris Newgard's lab, quantified and sent frozen in water. Six biological replicates were sent for the following conditions: (i) 1%BSA (12, 24, and 48 hours) (ii) 1mM-Oleate/Palmitate (12, 24, and 48 hours).

ORGANISM(S): Rattus norvegicus

SUBMITTER: Klaus Kaestner

PROVIDER: E-CBIL-34 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Functional genomics of the beta-cell: short-chain 3-hydroxyacyl-coenzyme A dehydrogenase regulates insulin secretion independent of K+ currents.

Hardy Olga T OT Hohmeier Hans E HE Becker Thomas C TC Manduchi Elisabetta E Doliba Nicolai M NM Gupta Rana K RK White Peter P Stoeckert Christian J CJ Matschinsky Franz M FM Newgard Christopher B CB Kaestner Klaus H KH

Molecular endocrinology (Baltimore, Md.) 20061221 3

Recent advances in functional genomics afford the opportunity to interrogate the expression profiles of thousands of genes simultaneously and examine the function of these genes in a high-throughput manner. In this study, we describe a rational and efficient approach to identifying novel regulators of insulin secretion by the pancreatic beta-cell. Computational analysis of expression profiles of several mouse and cellular models of impaired insulin secretion identified 373 candidate genes involv ...[more]

PMID: 17185391

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Project description:Obesity is becoming increasingly widespread in developed countries, and is often associated with heart diseases and diabetes. Elevated levels of plasma free fatty acids are a biochemical hallmark of obesity. Unlike plants and bacteria, mammals cannot utilize fatty acids to generate glucose because of the lack of glyoxylate shunt enzymes. Instead, fatty acids are used for energy storage, and their utilization is regulated at multiple levels ranging from hormonal to metabolic ensuring that glucose is preferentially oxidized before fatty acids. Here we designed a synthetic gene-metabolic circuit to alter the intracellular signaling and metabolic pathways that control fatty acid metabolism. By introducing the Escherichia coli glyoxylate shunt enzymes, isocitrate lyase (AceA) and malate synthase (AceB), into the mitochondria of human hepatocytes, we demonstrated that fatty acid utilization is preferentially increased while glucose uptake is significantly reduced. This bacterial pathway diverts signal metabolites (citrate, acetyl-CoA, and malonyl-CoA) that inhibits fatty acid uptake and provides an additional channel for fatty acid utilization. Remarkably, the hepatocytes readily adapted to the synthetic circuit by a series of global transcriptional and post-translational changes in metabolic and signal transduction pathways that further advanced our design objective. This systems approach illustrates the logic of the synergistic interaction between metabolism and signal transduction. The ready acceptance of this non-native pathway by hepatocytes suggests the plasticity of liver metabolism and opens a possibility for the synthetic approach in regulating human metabolism. Experiment Overall Design: Wild type (WT) human hepatocyte HepG2 and HepG2 cells stably transfected with pBudaceAB (designated ACE), a vector containing isocitrate lyase and malate synthase from E. coli, were analyzed after 24hour exposure to palmitate. Cells were seeded at 70% confluency in 10cm plates with DMEM growth media augmented with 300uM palmitate. After 24hours, total RNA was harvested and hybrized to Affymetrix HG_U133A 2.0 GeneChips. Data was processed using GCOS 1.2 software. Experiment Overall Design: 2 WT biological replicates and 2 ACE biological replicates were analyzed. Experiment Overall Design:

Dataset Information

Transcription profiling of rat insulinoma (INS-1 derived) cells grown in lipid culture

Publications

Functional genomics of the beta-cell: short-chain 3-hydroxyacyl-coenzyme A dehydrogenase regulates insulin secretion independent of K+ currents.

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