Unknown,Transcriptomics,Genomics,Proteomics

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Expression data from mock- or LMO3 silenced differentiating adipose stem cells

ABSTRACT: In this study we aimed to gain further insight on the role of GCs in adipocyte differentiation. For the future drugability of candidate targets it is of utmost importance to find factors relevant to human biology. Thus, we analyzed the transcriptome of GC induced primary human adipose stem cells (hASC) to identify novel factors downstream of GC action We used microarrays to detail the global programme of gene expression following glucocorticoid treatment and identified distinct classes of up- and downregulated genes during this process. Human preadipocytes (human adipose stem cells) were obtained from lipoaspirates by enzamytic digestions, followed by several steps of centrifugations (Mikkelsen et. al Cell. 2010 Oct 1;143(1):156-69.). Following isolation, human adipose stem cells were transfected with a control siRNA (siCtrl) or two different siRNA oligos targeting the gene LMO3 (siLMO3_oligo1 or siLMO3_oligo2). 40Hrs later, the transfected human adipose stem cells were induced to differentiate into mature adipocytes with a adipogenic cocktail (FullMix) and RNA isolated at day 0 or day6 after induction of differentiation.

ORGANISM(S): Homo sapiens

SUBMITTER: Markus Jeitler

PROVIDER: E-GEOD-41712 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background: Obesity is characterized as a disease that directly affects the whole-body metabolism and is associated with excess fat mass and several related comorbidities. Dynamics of the adipocyte hypertrophy and hyperplasia play an important role in health and disease, especially in obesity. Human adipose-derived stem cells (hASC) represent an important source for understanding the entire adipogenic differentiation process. However, little is known about the triggering step of adipogenesis in hASC. Here, we performed a proteogenomic approach for understanding the protein abundance alterations expression changes during the initiation of the adipogenic differentiation process. Methods: hASC were isolated from adipose tissue from three donors, characterized and expanded. Cells were cultured for 24 hours in adipogenic differentiation medium followed to protein extraction. We used shotgun proteomics to compare the proteomic profile of 24h-adipogenic differentiated and undifferentiated hASC. Besides, we used our previously next-generation sequencing data (RNA-seq) from the total and polysomal mRNA fractions of hASC to study post-transcriptional regulation during the initial steps of adipogenesis. Results: We identified a total of 3,.420 proteins out of and 48,.336 peptides, being 92 exclusively identified proteins in the undifferentiated hASC and 53 exclusive proteins in 24h-differentiated cells. Using a stringent criterion, we identified 33 differentially abundant proteins when compare 24h-differentiated versus undifferentiated hASC (14 upregulated and 19 downregulated, respectively). Among the upregulated proteins, we shortlist identified several adipogenic-related proteins. Combined analysis of the proteome and the transcriptome allowed the identification of positive correlation coefficients between proteins and mRNAs. Conclusions: These results demonstrate a specific proteome profile related to adipogenesis at the very beginning (24h) of the differentiation process in hASC, which represents an important piece for a better understanding of human adipogenesis and obesity. In addition, the adipogenic differentiation is fine-tuning regulated at transcriptional, post-transcriptional and post-translational levels.

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Expression data from mock- or LMO3 silenced differentiating adipose stem cells

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