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MRNA profiling of MEST knockdown in human adipocyte precursor cells at early stage (day 3) of adipocyte differentiation

ABSTRACT: The Mesoderm-specific transcript homolog protein MEST is a member of the alpha/beta hydrolase fold family that is expressed in adipose tissue, adipocytes as well as their precursor cells. To get an idea about potential functions of MEST in adipocyte development, we knocked down MEST in human Multipotent Adipose-Derived Stem (hMADS) cells, induced adipocyte differentiation by a chemically defined medium and performed gene expression profiling at the early stage of adipocyte differentiation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE63132 | GEO | 2015/06/24

SECONDARY ACCESSION(S): PRJNA266763

REPOSITORIES: GEO

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Project description:Abstract Background In obesity, adipose tissue undergoes a remodeling process characterized by increased adipocyte size (hypertrophia) and number (hyperplasia). The individual ability to tip the balance toward the hyperplastic growth, with recruitment of new fat cells through adipogenesis, seems to be critical for a healthy adipose tissue expansion, as opposed to the development of inflammation and detrimental metabolic consequences. However, the molecular mechanisms underlying this fine-tuned regulation are far from being understood. Methods We analyzed by mass spectrometry-based proteomics visceral white adipose tissue (vWAT) samples collected from C57BL6 mice fed with a HFD for 8 weeks. A subset of these mice, called low responders (LowR HFD), showed a low susceptibility to the onset of adipose tissue inflammation, as opposed to their HFD counterpart. We identified the discriminants between LowR HFD and HFD vWAT samples and explored their function in Adipose Derived human Mesenchymal Stem Cells (AD-hMSCs) differentiated to adipocytes. Results We quantified 6051 proteins. Among the candidates that most differentiate LowR HFD from HFD vWAT, we found proteins involved in adipocyte function, including adiponectin and hormone sensitive lipase, suggesting that adipocyte differentiation is enhanced in LowR HFD, as compared to HFD. The chromatin modifier SET and MYND Domain Containing 3 (SMYD3), whose function in adipose tissue was totally unknown, was another top-scored hit. SMYD3 expression was significantly higher in LowR HFD vWAT, as confirmed by western blot analysis. In vitro, we found that SMYD3 mRNA and protein levels decrease rapidly along the differentiation process of AD-hMSCs. Moreover, SMYD3 knock-down at the beginning of adipocyte differentiation resulted in reduced cell proliferation and, at longer term, reduced lipid accumulation in adipocytes. Conclusions Our study describes for the first time the role of SMYD3 as a regulator of adipocyte proliferation during the early steps of adipogenesis.

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MRNA profiling of MEST knockdown in human adipocyte precursor cells at early stage (day 3) of adipocyte differentiation

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