Multi‐omics analysis reveals the regulation of SIRT6 on protein processing of endoplasmic reticulum to alleviate oxidative stress in endothelial cells

Peng, Li‐Yan; Guo, Zhenyang; Feng, Runyang; Wu, Na; Zhong, Xin; Fang, Zheyan; Hu, Yiqing; Yu, Xiaohui; Zhao, Gang; He, Yue; Li, Hua; Ge, Junbo

doi:10.1002/ctm2.1039

Cited by 7 publications

(9 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(A) The correlation between Ero1α and SIRT6 in the proteome. (B) SIRT6 downregulated the expression of Ero1α in HUVECs exposed to 400 μM H 2 O 2 for 4 h. (The original data are from Figure 2H of our previously published article 15 . This is an open access article distributed under the terms of the Creative Commons CC BY license).…”

Section: Resultsmentioning

confidence: 99%

“…The detailed procedures for the preparation of cellular lysates and the Western blot analysis have been described previously 15 . The primary antibodies used in this study were as follows: HIF1α (36169S, CST, USA), Ero1α (3264S, CST, USA; 702709, Thermo Fisher, USA), p‐PERK (PA5‐102853, Thermo Fisher, USA), PERK (PA5‐120620, Thermo Fisher, USA), BIP (ab21685, Abcam, USA), p‐eif2α (3398S, CST, USA), eif2α (5324S, CST, USA), CHOP (2895S, CST, USA), CC3 (19677‐1‐AP, Proteintech, USA), Bax (ab32503, Abcam, USA), Bcl‐2(ab32124, Abcam, USA), SIRT6 (ab191385, Abcam, USA), p‐p65 (ab76302, USA), p65 (ab32536, USA), ICAM‐1 (ab53013, ab171123, Abcam, USA), VCAM‐1 (ab134047, Abcam, USA), VE‐cadherin (ab33168, Abcam, USA) and β‐actin (ab8226, Abcam, USA) was applied as the internal standard substances.…”

Section: Methodsmentioning

confidence: 99%

“…In addition, genetic ablation of SIRT6 in ECs could significantly increase the sensitivity of cells to stimuli, such as hypoxia, inflammation and oxidative stress, and intensify ECs dysfunctions and apoptosis 12 . In our previous work, SIRT6 was shown to alleviate ERS, reduce endothelial reactive oxygen species (ROS) levels and inhibit cellular apoptosis under oxidative stress 15 . However, the molecular mechanism by which SIRT6 regulates endothelial ERS during cardiac IRI remains unknown.…”

Section: Introductionmentioning

confidence: 99%

“… 12 In our previous work, SIRT6 was shown to alleviate ERS, reduce endothelial reactive oxygen species (ROS) levels and inhibit cellular apoptosis under oxidative stress. 15 However, the molecular mechanism by which SIRT6 regulates endothelial ERS during cardiac IRI remains unknown. In this study, SIRT6 was shown to protect CMECs against ERS‐induced dysfunction and apoptosis under IRI by repressing HIF1α and deacetylating H3K9 to inhibit Ero1α expression.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

SIRT6 deficiency in endothelial cells exacerbates oxidative stress by enhancing HIF1α accumulation and H3K9 acetylation at the Ero1α promoter

Guo,

Yu,

Zhao

et al. 2023

Clinical & Translational Med

Self Cite

View full text Add to dashboard Cite

BackgroundSIRT6, an important NAD+‐dependent protein, protects endothelial cells from inflammatory and oxidative stress injuries. However, the role of SIRT6 in cardiac microvascular endothelial cells (CMECs) under ischemia‒reperfusion injury (IRI) remains unclear.MethodsThe HUVECs model of oxygen–glucose deprivation/reperfusion (OGD/R) was established to simulate the endothelial IRI in vitro. Endoplasmic reticulum oxidase 1 alpha (Ero1α) mRNA and protein levels in SIRT6‐overexpressing or SIRT6‐knockdown cells were measured by qPCR and Western blotting. The levels of H2O2 and mitochondrial reactive oxygen species (ROS) were detected to evaluate the status of oxidative stress. The effects of SIRT6 deficiency and Ero1α knockdown on cellular endoplasmic reticulum stress (ERS), inflammation, apoptosis and barrier function were detected by a series of molecular biological experiments and functional experiments in vitro. Chromatin immunoprecipitation, Western blotting, qPCR, and site‐specific mutation experiments were used to examine the underlying molecular mechanisms. Furthermore, endothelial cell‐specific Sirt6 knockout (ecSirt6−/−) mice were subjected to cardiac ischemia‒reperfusion surgery to investigate the effects of SIRT6 in CMECs in vivo.ResultsThe expression of Ero1α was significantly upregulated in SIRT6‐knockdown endothelial cells, and high Ero1α expression correlated with the accumulation of H2O2 and mitochondrial ROS. In addition, SIRT6 deficiency increased ERS, inflammation, apoptosis and endothelial permeability, and these effects could be significantly attenuated by Ero1α knockdown. The deacetylase catalytic activity of SIRT6 was important in regulating Ero1α expression and these biological processes. Mechanistically, SIRT6 inhibited the enrichment of HIF1α and p300 at the Ero1α promoter through deacetylating H3K9, thereby antagonizing HIF1α/p300‐mediated Ero1α expression. Compared with SIRT6‐wild‐type (SIRT6‐WT) cells, cells expressing the SIRT6‐H133Y‐mutant and SIRT6‐R65A‐mutant exhibited increased Ero1α expression. Furthermore, ecSirt6−/− mice subjected to ischemia‒reperfusion surgery exhibited increased Ero1α expression and ERS in CMECs and worsened injuries to microvascular barrier function and cardiac function.ConclusionsOur results revealed an epigenetic mechanism associated with SIRT6 and Ero1α expression and highlighted the therapeutic potential of targeting the SIRT6‐HIF1α/p300‐Ero1α axis.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

SIRT6 deficiency in endothelial cells exacerbates oxidative stress by enhancing HIF1α accumulation and H3K9 acetylation at the Ero1α promoter

Guo,

Yu,

Zhao

et al. 2023

Clinical & Translational Med

Self Cite

View full text Add to dashboard Cite

show abstract

“…The molecular interaction between Sirt6 and AIM2-pyroptosis forms a complex signal transduction pathway to control the occurrence and outcome of inflammation. Sirt6 is indicated to be an endogenous inhibitive regulator of oxidative stress [ 42 ], and oxidative stress directly activate the AIM2 and its downstream signaling pathway [ 43 ]. AIM2 pattern recognition receptor binds to infectious and/or non-infectious stimuli, which activates the downstream multi-protein inflammasome complex, and then promotes caspase-1 dependent GSDMD to locate on cell membrane, forming pores conducive to the release of IL-1β and IL-18, and finally leads to pyroptosis [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

Exploring the therapeutic potential of Sirt6-enriched adipose stem cell-derived exosomes in myocardial ischemia–reperfusion injury: unfolding new epigenetic frontiers

Liu,

Wang,

Wang

et al. 2024

Clin Epigenet

View full text Add to dashboard Cite

Background The management of myocardial ischemia–reperfusion injury (MIRI) presents continuous therapeutic challenges. NAD-dependent deacetylase Sirtuin 6 (Sirt6) plays distinct roles in various disease contexts and is hence investigated for potential therapeutic applications for MIRI. This study aimed to examine the impact of Sirt6-overexpressing exosomes derived from adipose stem cells (S-ASC-Exo) on MIRI, focusing on their influence on AIM2-pyroptosis and mitophagy processes. The sirtuin family of proteins, particularly Sirtuin 6 (Sirt6), play a pivotal role in these processes. This study aimed to explore the potential therapeutic effects of Sirt6-enriched exosomes derived from adipose stem cells (S-ASC-Exo) on regulating MIRI. Results Bioinformatic analysis revealed a significant downregulation of Sirt6 in MIRI subjected to control group, causing a consequential increase in mitophagy and pyroptosis regulator expressions. Therefore, our study revealed that Sirt6-enriched exosomes influenced the progression of MIRI through the regulation of target proteins AIM2 and GSDMD, associated with pyroptosis, and p62 and Beclin-1, related to mitophagy. The introduction of S-ASC-Exo inhibited AIM2-pyroptosis while enhancing mitophagy. Consequently, this led to a significant reduction of GSDMD cleavage and pyroptosis in endothelial cells, catalyzing a deceleration in the progression of atherosclerosis. Extensive in vivo and in vitro assays were performed to validate the expressions of these specific genes and proteins, which affirmed the dynamic modulation by Sirt6-enriched exosomes. Furthermore, treatment with S-ASC-Exo drastically ameliorated cardiac functions and limited infarct size, underlining their cardioprotective attributes. Conclusions Our study underscores the potential therapeutic role of Sirt6-enriched exosomes in managing MIRI. We demonstrated their profound cardioprotective effect, evident in the enhanced cardiac function and attenuated tissue damage, through the strategic modulation of AIM2-pyroptosis and mitophagy. Given the intricate interplay between Sirt6 and the aforementioned processes, a comprehensive understanding of these pathways is essential to fully exploit the therapeutic potential of Sirt6. Altogether, our findings indicate the promise of Sirt6-enriched exosomes as a novel therapeutic strategy in treating ischemia–reperfusion injuries and cardiovascular diseases at large. Future research needs to underscore optimizing the balance of mitophagy during myocardial ischemia to avoid potential loss of normal myocytes.

show abstract

Targeting endoplasmic reticulum stress‐induced lymphatic dysfunction for mitigating bisphosphonate‐related osteonecrosis

Qin,

Xie,

et al. 2024

Clinical & Translational Med

View full text Add to dashboard Cite

BackgroundBisphosphonates (BPs) are the first‐line treatment to stop bone resorption in diseases, including osteoporosis, Paget's disease, multiple myeloma and bone metastases of cancer. However, BPs‐related osteonecrosis of the jaw (BRONJ), characterized by local inflammation and jawbone necrosis, is a severe intractable complication. The cumulative inflammatory burden often accompanies impaired lymphatic drainage, but its specific impact on BRONJ and the underlying mechanisms remain unclear.MethodsThe mouse BRONJ model was established to assess the integrity and drainage function of lymphatic vessels by tissue clearing techniques, injected indocyanine green lymphatic clearance assay, flow cytometry analysis and histopathological staining. RNA sequencing, metabolome analysis, transmission electron microscopy and Western blotting were utilized to analyze the impacts of Zoledronate acid (ZA) on endoplasmic reticulum stress (ERS) and function of lymphatic endothelial cells (LECs). By constructing Lyve1creERT; SIRT6f/f and Lyve1creERT; ATG5f/f mice, we evaluated the role of ERS‐induced LECs apoptosis in the progression of BRONJ. Additionally, we developed a nanoparticle‐loaded ZA and rapamycin (ZDPR) to enhance autophagy and evaluated its potential in mitigating BRONJ.ResultsThe mouse BRONJ model displayed impaired lymphatic drainage, accompanied by significant local inflammation and bone necrosis. The prolonged stimulation of ZA resulted in the extension of ERS and the inhibition of autophagy in LECs, ultimately leading to apoptosis. Mechanistically, ZA activated XBP1s through the NAD+/SIRT6 pathway, initiating ERS‐induced apoptosis in LECs. The conditional knockout mouse models demonstrated that the deletion of SIRT6 or ATG5 significantly worsened lymphatic drainage and inflammatory infiltration in BRONJ. Additionally, the innovative nanoparticle ZDPR alleviated ERS‐apoptosis in LECs and enhanced lymphatic function, facilitating inflammation resolution.ConclusionOur study has elucidated the role of the NAD+/SIRT6/XBP1s pathway in ERS‐induced apoptosis in ZA‐treated LECs, and further confirmed the therapeutic potential of ZDPR in restoring endothelial function and improving lymphatic drainage, thereby effectively mitigating BRONJ.Key points Bisphosphonate‐induced lymphatic drainage impairment exacerbates bone necrosis. Zoledronate acid triggers endoplasmic reticulum stress and apoptosis in lymphatic endothelial cells via the NAD+/SIRT6/XBP1s pathway. Novel nanoparticle‐loaded Zoledronate acid and rapamycin enhances autophagy, restores lymphatic function, and mitigates bisphosphonates‐related osteonecrosis of the jaw progression.

show abstract

Multi‐omics analysis reveals the regulation of SIRT6 on protein processing of endoplasmic reticulum to alleviate oxidative stress in endothelial cells

Cited by 7 publications

References 8 publications

SIRT6 deficiency in endothelial cells exacerbates oxidative stress by enhancing HIF1α accumulation and H3K9 acetylation at the Ero1α promoter

SIRT6 deficiency in endothelial cells exacerbates oxidative stress by enhancing HIF1α accumulation and H3K9 acetylation at the Ero1α promoter

Exploring the therapeutic potential of Sirt6-enriched adipose stem cell-derived exosomes in myocardial ischemia–reperfusion injury: unfolding new epigenetic frontiers

Targeting endoplasmic reticulum stress‐induced lymphatic dysfunction for mitigating bisphosphonate‐related osteonecrosis

Contact Info

Product

Resources

About