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The Effect of Weight Loss on Metabolic Processes of the Abdominal Subcutaneous Adipose Tissue: An Exploratory Study using total RNA Next Generation Sequencing

ABSTRACT: Obesity induces profound alterations in adipose tissue biology, yet the extent to which these transcriptional changes resolve after weight loss remains unclear. This exploratory study investigated molecular remodeling of abdominal subcutaneous adipose tissue (SAT) in formerly obese individuals.

ORGANISM(S): Homo sapiens

PROVIDER: GSE329809 | GEO | 2026/05/07

REPOSITORIES: GEO

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Project description:Objective: This work aimed at identifying and characterizing differences in subcutaneous adipose tissue (SAT) gene expression patterns between subjects succeeding weight control against subjects regaining weight 6 months after a low calorie diet program. Methods: Weight-reduced obese subjects from 8 European countries were randomized into 4 diets differing in protein and glycemic index. In addition to the anthropometric and plasma parameters, SAT biopsies were taken at the beginning (CID2) and the end (CID3) of the weight maintenance intervention. Pan-genomic cDNA microarrays were used to define SAT gene expression profiles at both CID2 and CID3 of 22 subjects succeeding weight maintenance (successful subjects; mean weight change= -2.6 ± 1.2 kg) and 22 subjects regaining weight (unsuccessful subjects; mean weight change= 3.9 ± 1.3 kg) among the 4 dietary arms. Results: Differences in SAT gene expression patterns between successful and unsuccessful groups were mainly due to weight variations rather to differences in diet macronutrient composition. An ANCOVA analysis with total energy intake at CID3 as covariant led us to the definition of 1339 differential genes. Functional analysis of differential genes showed that cellular growth and proliferation, inflammation, cell death, cancer, cellular function and maintenance were the main biological processes involving these differential genes. Overall design: SAT transcriptome was defined at the beginning (CID2) and at the end (CID3) of the dietary intervention program using a common reference design: Cyanine-5 dye was incorporated into all SAT samples, while a reference RNA pool made of the mix of commercial human liver, adipose tissue, and skeletal muscle RNA was labeled with cyanine-3 dye. Samples were hybridized to Agilent 44K whole human genome microarrays. Evolution in gene expression patterns after dietary intervention was tested in terms of the ratio after/before dietary intervention (CID3/CID2) ratio.

Project description:Objective: Abdominal adiposity is strongly associated with diabetic and cardiovascular comorbidities. The long non-coding RNA HOTAIR (HOX Transcript Antisense Intergenic RNA) is an important epigenetic regulator, with fat depot-specific expression between abdominal subcutaneous adipose tissue (SAT) and gluteal SAT. HOTAIR locates closely with HOXC13, known to strongly associated with human fat distribution. Here, we examined the phenotypic effects of HOTAIR overexpression on abdominal adipogenesis, and hypothesized that HOTAIR-mediated DNA methylation is correlated with transcriptome changes, leading to the regulation of specific genes and the functional pathways. Methods: The expression level of HOTAIR was compared among different fat-depots collected from six healthy, five severe obese, and five uremic subjects, and was correlated with dual-energy x-ray absorptiometry (DXA) defined regional adiposity. The human immortalized preadipocyte was used to assess the phenotypic effects of HOTAIR overexpression on abdominal adipogenesis. The integrative analysis of reduced representation bisulfite sequencing (RRBS) and RNA-sequencing was performed to identify putative genes that are epigenetically regulated by HOTAIR, and the associated signaling pathways. HOTAIR-repressed genes were further validated using RNA/chromatin immunoprecipitation with real-time qPCR and correlated with human body fat distribution. Results: We found that the expression of HOTAIR was high in gluteal SAT, and low in arm/abdominal SAT and visceral (omental) adipose tissue. It could be aberrantly increased in uremic arm SAT. Notably, in severe obese subjects we found that HOTAIR is lowly expressed in abdominal SAT correlating with a higher abdominal adiposity, whereas in uremic patients HOTAIR is highly expressed in arm SAT correlating with lower arm adiposity. HOTAIR overexpression in human immortalized abdominal preadipocyte remarkably suppresses the in vitro adipogenesis. Overall the differentially methylated genes were functionally enriched for nervous system development. We specifically identified 10 HOTAIR-mediated genes showing strong changes of DNA methylation associated with gene expression during abdominal adipogenesis, suggesting potential epigenetic regulation. Two HOTAIR-repressed genes, SLITRK4 and PITPNC1, were further highlighted and validated; presenting an obesity-driven fat-depot specific expression pattern positively correlated with the central body fat distribution. Conclusions: Our study indicated that HOTAIR is an important regulator for abdominal adipogenesis via intricate DNA methylation likely to associate with transcriptional regulation of specific genes, such as SLITRK4 and PITPNC1.

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The Effect of Weight Loss on Metabolic Processes of the Abdominal Subcutaneous Adipose Tissue: An Exploratory Study using total RNA Next Generation Sequencing

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