Lnc13728 facilitates human mesenchymal stem cell adipogenic differentiation via positive regulation of ZBED3 and downregulation of the WNT/β-catenin pathway

Abstract

Background Obesity has received increasing attention because of its widespread worldwide occurrence and many threats to health. Human adipose-derived mesenchymal stem cells (hADSCs) are a critical source of adipocytes. Long noncoding RNAs (lncRNAs) play pivotal roles in cell fate determination and differentiation. The objective of the present study was to identify and investigate the function and regulatory mechanism of lncRNAs on adipogenic differentiation of hADSCs. Methods We used lncRNA arrays to identify the prominent differentially expressed lncRNAs before and after hADSC adipogenic differentiation and verified their biological function through antisense oligonucleotide knockdown or lentivirus overexpression. The adipogenic differentiation of hADSCs was assessed by oil red O staining as well as the mRNA and protein levels of adipogenic marker genes through qRT-PCR and western blot. Bioinformatic tool LncPro and immunofluorescence was performed to uncover the interaction between lnc13728 and ZBED3. WNT/β-catenin signaling pathway was evaluated by western blot and immunofluorescence. Results The lncRNA arrays showed that lnc13728 expression was significantly upregulated after hADSC adipogenic differentiation and was correlated positively with the expression of the adipogenesis-related genes in human adipose tissue. Lnc13728 knockdown in hADSCs suppressed the expression of the adipogenesis-related genes at both mRNA and protein level and weakened lipid droplet production. Accordingly, lnc13728 overexpression enhanced hADSC adipogenic differentiation. Beyond that, lnc13728 co-localized with ZBED3 in the cytoplasm and regulated its expression positively. Downregulating ZBED3 had a negative effect on adipogenic differentiation, while the expression of WNT/β-catenin signaling pathway-related proteins was upregulated. Conclusions Lnc13728 promotes hADSC adipogenic differentiation possibly by positively regulating the expression of ZBED3 which plays a role in inhibiting the WNT/β-catenin pathway.

Long Non-coding RNA Regulation of Mesenchymal Stem Cell Homeostasis and Differentiation: Advances, Challenges, and Perspectives

“…Another newly identified lncRNA, PGC1β-OT1, reciprocally modulates MSC adipogenic and osteogenic commitment by sponging miR-148a-3p and enhancing the effect of KDM6B (Yuan et al, 2019); the lncRNA ROA inhibits MSC adipogenic differentiation by destroying hnRNPA1 binding to the PTX3 promoter, thereby transcriptionally downregulating PTX3 and the ERK pathway (Pan et al, 2020). Moreover, our lab discovered that lncRNA13728 promoted ADSC adipogenic differentiation by upregulating ZBED3 and inhibiting the WNTβ-catenin pathway (Xu et al, 2021).…”

Epigenomic and transcriptomic prioritization of candidate obesity-risk regulatory GWAS SNPs

“…Thus, these candidates for regulatory SNPs are likely to be relevant to obesity-related disease susceptibility (3). Specific roles in adipose biology have been inferred for the seven gene loci associated with these SNPs (3,5,25,39,44,(52)(53)(54)(55)(56)(57)(58). Remarkably, four of these genes, CABLES1, PEMT, PC, and FAM13A, had one or more of their Tier-1 SNPs near an alternate tissue-specific TSS.…”

“…These miRNAs are implicated in nonalcoholic fatty liver disease (67) or gene dysregulation in VAT in morbidly obese individuals (68). ZBED3-AS1 displays increases in its steady-state RNA levels during in vitro adipogenesis and levels of this RNA correlate with expression of adipogenesis markers in liposuction samples (44). Moreover, production of ZBED3-AS1 RNA was reported to be necessary for normal levels of expression of markers of adipogenesis during in vitro differentiation (44).…”

“…ZBED3-AS1 might influence expression in SAT of several genes in its 1-Mb neighborhood, including ZBED3, which encodes Zinc Finger BED Domain-Containing Protein 3 and is oriented head-to-head with ZBED3-AS1 with a 0.5 kb overlap. ZBED3 and ZBED3-AS1 have been implicated in adipogenesis (44). The ZBED3-AS1 Tier-1 SNPs are in low LD with each other (r 2 = 0.2, EUR) and reside 5 kb apart in separate enhancer chromatin segments and DNase hypersensitivity peaks in SAT (Figure 5B-D).…”

RIPK1 Coordinates Bone Marrow Mesenchymal Stem Cell Survival by Maintaining Mitochondrial Homeostasis via p53

“…Then, cells were fixed, washed, and stained in Oil Red O dye solution (Beyotime Biotech, China) according to the manufacturer's protocol. After incubation for 30 min, the cells were washed with staining buffer and photographed by a microscope [27].…”