Permanent cystathionine-β-Synthase gene knockdown promotes inflammation and oxidative stress in immortalized human adipose-derived mesenchymal stem cells, enhancing their adipogenic capacity

Abstract

In the present study, we aimed to investigate the impact of permanent cystathionine-β-Synthase (CBS) gene knockdown in human telomerase reverse transcriptase (hTERT) immortalized human adipose-derived mesenchymal stem cells (ASC52telo) and in their capacity to differentiate into adipocytes. CBS gene KD in ASC52telo cells led to increased cellular inflammation ( IL6, CXCL8, TNF ) and oxidative stress markers (increased intracellular reactive oxygen species and decreased reduced glutathione levels) in parallel to decreased H 2 S production and rejuvenation ( LC3 and SIRT1 )-related gene expression. In addition, CBS gene KD in ASC52telo cells resulted in altered mitochondrial respiratory function, characterised by decreased basal respiration (specifically proton leak) and spare respiratory capacity, without significant effects on cell viability and proliferation. In this context, shCBS-ASC52telo cells displayed enhanced adipogenic ( FABP4, ADIPOQ, SLC2A4, CEBPA, PPARG )-, lipogenic ( FASN, DGAT1 )- and adipocyte ( LEP, LBP )-related gene expression markers, decreased expression of proinflammatory cytokines, and increased intracellular lipid accumulation during adipocyte differentiation compared to control ASC52telo cells. Otherwise, the increased adipogenic potential of shCBS-ASC52telo cells was detrimental to the ability to differentiate into osteogenic linage. In conclusion, this study demonstrated that permanent CBS gene KD in ASC52telo cells promotes a cellular senescence phenotype with a very increased adipogenic potential, promoting a non-physiological enhanced adipocyte differentiation with excessive lipid storage.

Activation of Endogenous H2S Biosynthesis or Supplementation with Exogenous H2S Enhances Adipose Tissue Adipogenesis and Preserves Adipocyte Physiology in Humans

“…In line with these findings, in immortalized human adipose-derived mesenchymal stem cells, which unlike human preadipocytes and adipocytes (current study) displayed a much higher expression of CBS than CTH gene (18), CBS gene KD promotes a cellular senescence phenotype characterized by increased inflammation and oxidative stress, and decreased H 2 S production (18). Of note, this cellular senescence phenotype resulted in adipocyte hypertrophy, when these cells differentiated into adipocytes, and attenuated their ability to differentiate into osteogenic linage (18).…”

“…Strengthening current findings, increased SIRT1 activity led to enhanced adipose tissue rejuvenation (characterized by increased cellular stemness) and decreased inflammation (12,29,43,64,78) and H 2 S administration resulted in enhanced Sirt1 expression and activity (20,51,65), preventing vascular aging (20) and cellular senescence in human fibroblasts (65). In line with these findings, in immortalized human adipose-derived mesenchymal stem cells, which unlike human preadipocytes and adipocytes (current study) displayed a much higher expression of CBS than CTH gene (18), CBS gene KD promotes a cellular senescence phenotype characterized by increased inflammation and oxidative stress, and decreased H 2 S production (18). Of note, this cellular senescence phenotype resulted in adipocyte hypertrophy, when these cells differentiated into adipocytes, and attenuated their ability to differentiate into osteogenic linage (18).…”

Single-cell transcriptomic analysis of eutopic endometrium and ectopic lesions of adenomyosis

“…Compared with the AM_EM sample, the AM_EC sample was enriched for cell motility and inflammation-related terms according to the GO and KEGG results, and the volcano map results showed that associated gene transcription, including that of ITGA2 [ 27 ], ITGB8 [ 41 ] and CXCL8 [ 42 ], was upregulated. Since the pathogenesis of AM is closely related to the functional changes in the endometrium [ 43 ], we identified the functional changes in endometrial epithelial cells in the AM_EC and AM_EM groups and found that several identical items related to cell movement and inflammation, including positive regulation of cell migration and the IL-17 signalling pathway, appeared in the comparison of the AM_EC versus AM_EM groups in epithelial cells.…”

Vitamin B12 Attenuates Acute Pancreatitis by Suppressing Oxidative Stress and Improving Mitochondria Dysfunction via CBS/SIRT1 Pathway

“…Cysteine- β -synthase (CBS) is widely distributed in the liver, kidney, and pancreas and is the first (and rate-limiting) enzyme in the GSH synthesis pathway [ 6 , 7 ]. A recent study showed CBS gene knockdown promotes inflammation and oxidative stress in immortalized human adipose-derived mesenchymal stem cells, enhancing their adipogenic capacity [ 8 ]. In addition, the decreased expression of CBS propagates the pathogenesis of ulcerative colitis by exacerbating inflammation-induced intestinal barrier injury [ 9 ], implying an important role of CBS in mediating organ damage.…”