Caveolin-1 rs1997623 Single Nucleotide Polymorphism Creates a New Binding Site for the Early B-Cell Factor 1 That Instigates Adipose Tissue CAV1 Protein Overexpression.
Ashraf Al MadhounDania HaddadRasheeba NizamLavina MirandaShihab KochumonReeby ThomasThangavel Alphonse ThanarajRasheed AhmadMilad S BitarFahd Al-MullaPublished in: Cells (2022)
Caveolin-1 (CAV1) is implicated in the pathophysiology of diabetes and obesity. Previously, we demonstrated an association between the CAV1 rs1997623 C > A variant and metabolic syndrome (MetS). Here, we decipher the functional role of rs1997623 in CAV1 gene regulation. A cohort of 38 patients participated in this study. The quantitative MetS scores (siMS) of the participants were computed. CAV1 transcript and protein expression were tested in subcutaneous adipose tissue using RT-PCR and immunohistochemistry. Chromatin immunoprecipitation assays were performed using primary preadipocytes isolated from individuals with different CAV1 rs1997623 genotypes (AA, AC, and CC). The regulatory region flanking the variant was cloned into a luciferase reporter plasmid and expressed in human preadipocytes. Additional knockdown and overexpression assays were carried out. We show a significant correlation between siMS and CAV1 transcript levels and protein levels in human adipose tissue collected from an Arab cohort. We found that the CAV1 rs1997623 A allele generates a transcriptionally active locus and a new transcription factor binding site for early B-cell factor 1 (EBF1), which enhanced CAV1 expression. Our in vivo and in vitro combined study implicates, for the first time, EBF1 in regulating CAV1 expression in individuals harboring the rs1997623 C > A variant.
Keyphrases
- adipose tissue
- transcription factor
- metabolic syndrome
- insulin resistance
- type diabetes
- poor prognosis
- endothelial cells
- end stage renal disease
- high fat diet
- binding protein
- cell proliferation
- crispr cas
- ejection fraction
- small molecule
- chronic kidney disease
- high throughput
- body mass index
- dna damage
- rna seq
- high resolution
- computed tomography
- oxidative stress
- dna methylation
- skeletal muscle
- peritoneal dialysis
- prognostic factors
- genome wide
- induced pluripotent stem cells
- dna binding
- single cell
- cardiovascular risk factors
- high throughput sequencing