We demonstrate how a multidisciplinary approach encompassing cellular reprogramming, transcriptomics, proteomics, metabolomics, modeling, network reconstruction and data management can be employed to unveil the mechanisms underlying the progression of steatosis.
More...We demonstrate how a multidisciplinary approach encompassing cellular reprogramming, transcriptomics, proteomics, metabolomics, modeling, network reconstruction and data management can be employed to unveil the mechanisms underlying the progression of steatosis. Proteomics revealed reduced AKT/mTOR signaling in fibroblasts derived from steatosis patients and further establishes that the insulin-resistant phenotype is present not only in insulin-metabolizing central organs, e.g. the liver, but is also manifested in skin fibroblasts. Transcriptome data enabled the generation of a regulatory network based on the transcription factor SREBF1, linked to a metabolic network of glycerolipid and fatty acid biosynthesis including the downstream transcriptional targets of SREBF1 which include LIPIN1 (LPIN) and low density lipoprotein receptor (LDLR). Glutathione metabolism was among the pathways enriched in steatosis patients in comparison to healthy controls. By using a model of the glutathione pathway we predict a significant increase in the flux through glutathione synthesis as both gamma-glutamylcysteine synthetase and glutathione synthetase have an increased flux. The transcriptomes of dermal fibroblasts derived from two patients with steatosis (60 yr, male BMI 45.7 (H0008) and 33 yr, female, BMI 51.3 (H0007)) and control fibroblasts (19 yr, female, BMI 21, (H0002)) were analyzed.
Overall design: Total RNA obtained from dermal fibroblasts derived from two patients with steatosis (60 yr, male BMI 45.7 (H0008) and 33 yr, female, BMI 51.3 (H0007)) and control fibroblasts (19 yr, female, BMI 21, (H0002)).
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