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Functional and comparative analysis of SWAT cells

ABSTRACT: Human adipose stem and progenitor cells (ASPCs) develop into heterogenous cultures of adipogenic and Structural Wnt-regulated Adipose Tissue resident (SWAT) cells upon induction of adipogenic differentiation. To investiagte the function of SWAT cells and compare them with proliferating ASPCs, we enriched the SWAT cells from the adipogenic cells using density gradient centriguation. The enriched SWAT cells were cultured for 24 hours in proliferation media or differentiation media for 24 hours. 4 density gradient centrifugations were performed, resulting in 4 technical replicates per media type. We included proliferating ASPCS (undifferentiated) as control. We then performed gene expression profiling analysis using data from RNA-Seq of 4 samples per condition (proliferating ASPCs, SWAT cells in proliferation media, SWAT cells in differentiation media)

ORGANISM(S): Homo sapiens

PROVIDER: GSE223588 | GEO | 2023/04/03

REPOSITORIES: GEO

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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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Functional and comparative analysis of SWAT cells

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