EUROPEAN JOURNAL OF PHARMACOLOGY
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2024
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IF 4.2
Liu-Gen Cui,Miao-Miao Zhai,Jian-Jian Yin,Zhi-Mo Wang,Shu-Hui Wang,Yue-Jiao Zhou,Pei-Pei Li,Yang Wang,Li Xia,Pei Wang,Xue-Xiang Cha,Li-Rong Zhang,Sheng-Na Han
Several studies have associated the epitranscriptomic RNA modification of N6-methyladenosine (m 6 A) with cardiovascular diseases; however, how m 6 A modification affects cardiomyocyte pyroptosis after myocardial infarction (MI) remains unknown. Here, we showed that AlkB homolog 5 (ALKBH5), an m 6 A demethylase, is crucial in cardiomyocyte pyroptosis after MI. We used MI rat and mouse models, a cell hypoxia model of rat primary cardiomyocytes (RCMs), and rat embryonic ventricle cell line (H9c2) to explore the functional role of m 6 A modification and ALKBH5 in the heart and cardiomyocytes. Using plasmids and small interfering RNAs, the expressions of ALKBH5 and NOD-like receptor family pyrin domain-containing 3 (NLRP3) were determined to study their functions in regulating cardiomyocyte m 6 A and pyroptosis, respectively. We characterized the role of ALKBH5, which exhibited elevated expression in the ischemic heart tissue of rats and mice and hypoxic cardiomyocytes (RCMs and H9c2 cells). ALKBH5 knockdown alleviated hypoxia-induced H9c2 cell pyroptosis by inhibiting NLRP3 inflammasome activation, whereas ALKBH5 overexpression had the opposite effect. NLRP3 knockdown alleviated hypoxia-induced H9c2 cardiomyocyte pyroptosis by inhibiting ALKBH5 expression, whereas NLRP3 overexpression had the opposite effect. Mechanistically, ALKBH5 mediated m 6 A modification of NLRP3 mRNA in an IGF2BP2-dependent manner, and NLRP3, as a nuclear transcription factor, regulated the ALKBH5 transcription process. Targeting the ALKBH5-NLRP3 loop with the small-molecule inhibitors alleviated cardiomyocyte pyroptosis. Our results highlight that ALKBH5-NLRP3 forms a positive feedback loop that promotes cardiomyocyte pyroptosis after MI. Therefore, inhibiting the ALKBH5-NLRP3 loop is a potential strategy for treating cardiovascular diseases.
研究领域:
心血管疾病
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表观转录组学
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分子生物学
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高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2021
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IF 6.291
Luyao Li,Mei Zhou,Biyun Chen,Qin Wang,Shuya Pan,Yaxuan Hou,Jiamin Xia,Xue Zhou
Cadmium is a carcinogenic heavy metal that poses a severe threat to human beings. The underlying mechanism, however, remains elusive. N6-methyladenosine (m6A) is the most abundant post-transcriptional modification in mRNA that regulates RNA metabolism. Emerging evidence shows that m6A is involved in the pathogenesis of various cancers. In this study, human bronchial epithelial BEAS-2B cells were transformed by exposing to 2 μM of cadmium for 20 weeks to investigate the role of m6A in cadmium carcinogenesis. We found the level of m6A in mRNA was significantly decreased in cadmium-transformed BEAS-2B cells, and this change was regulated by m6A demethylase ALKBH5. ALKBH5 was significantly upregulated in the middle and late stages of cell transformation at week 8, 12, 16 and 20. Knockdown of ALKBH5 in cadmium-transformed cells alleviated cell proliferation, migration, invasion, and anchorage-independent growth, but co-transfection with ALKBH5 siRNA and PTEN siRNA restored the inhibitory effects of ALKBH5 knockdown on those transformation properties. ALKBH5 decreased the m6A level of PTEN mRNA, resulting in its instability and reduction of PTEN protein expression. These results indicate that ALKBH5-mediated demethylation m6A at PTEN mRNA is involved in cadmium-induced cell transformation. Our study provides a new perspective for the involvement of m6A modification in cadmium carcinogenesis.
研究领域:
毒理学
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分子生物学
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癌症研究
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PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2024
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IF 12.2
Se-Ruo Li,Ning-Ning Kang,Rong-Rong Wang,Meng-Die Li,Li-Hong Chen,Peng Zhou,De-Xiang Xu,Hui Zhao,Lin Fu
Our previous study revealed that 1-nitropyrene (1-NP) exposure evoked pulmonary fibrosis in mice. However, the exact mechanism remained elusive. We found that 1-NP induced telomere damage and cellular senescence in mice lungs, and two alveolar epithelial cells lines. 1-NP downregulated telomere repeat binding factor 2 (TRF2), and upregulated FBXW7. Mechanistically, 1-NP-caused TRF2 ubiquitination and proteasomal degradation depended on E3 ubiquitin ligase activity of FBXW7. Moreover, 1-NP upregulated FBXW7 m6A modification via an ALKBH5-YTHDF1-dependent manner. Further analysis suggested 1-NP promoted ALKBH5 SUMOylation and subsequent proteasomal degradation. Additionally, 1-NP evoked mitochondrial reactive oxygen species (mtROS) overproduction. Mito-TEMPO, a mitochondrial-targeted antioxidant, mitigated 1-NP-caused mtROS overproduction, ALKBH5 SUMOylation, FBXW7 m6A modification, TRF2 degradation, cellular senescence, and pulmonary fibrosis. Taken together, mtROS-initiated ALKBH5 SUMOylation and subsequent FBXW7 m6A modification is indispensable for TRF2 degradation and cellular senescence in alveolar epithelial cells during 1-NP-induced pulmonary fibrosis. Our study provides target intervention measures towards 1-NP-evoked pulmonary fibrosis. Environmental Implication 1-nitropyrene (1-NP) is a representative of nitro polycyclic aromatic hydrocarbon compound that can be formed during the insufficient combustion of organic substances like coal, oil, gas, and tobacco. Moreover, mounting researches have revealed that 1-NP can cause various respiratory system diseases, inclusive of acute lung injury, chronic obstructive pulmonary disease, and allergic asthma. The current research provides evidence that exposure to 1-NP induces pulmonary fibrosis through evoking telomere damage and cellular senescenc. What’s more, Mito-TEMPO, a mitochondrial-targeted antioxidant, can mitigate 1-NP-caused cellular senescence and pulmonary fibrosis. Our study provides target intervention measures towards 1-NP-evoked pulmonary fibrosis.
研究领域:
毒理学
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肺科学
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细胞生物学
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分子生物学
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环境健康
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高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2023
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IF 13.6
Weiwei Wang,Xin Li,Qiuhui Qian,Jin Yan,Haishan Huang,Xuedong Wang,Huili Wang
Because triclosan (TCS) has been confirmed to cause severe neurotoxicity, it is urgent to disclose the underlying toxicity mechanisms at varying levels. TCS exposure resulted in a series of malformations in larval zebrafish, including reduced neurons, blood-vessel ablation and abnormal neurobehavior. Apoptosis staining and the upregulated expression of proapoptotic genes demonstrated that TCS induced neuronal apoptosis and neurotransmitter disorders. By integrating RT-qPCR analysis with the effects of pathway inhibitors and agonists, we found that TCS triggered abnormal regulation of neuron development-related functional genes, and suppressed the BDNF/TrkB signaling pathway. TCS inhibited total m 6 A-RNA modification level by activating the demethylase ALKBH5, and induced neurodevelopmental toxicity based on the knockdown experiments of alkbh5 and molecular docking . The main novelties of this study lies in: (1) based on specific staining and transgenic lines, the differential neurotoxicity effects of TCS were unravelled at individual, physiological, biochemical and molecular levels in vivo ; (2) from a epigenetics viewpoint, the decreasing m 6 A methylation level was confirmed to be mediated by alkbh5 upregulation; and (3) both homology modeling and molecular docking evidenced the targeting action of TCS on ALKBH5 enzyme. These findings open a novel avene for TCS’s risk assessment and early intervention of the contaminant-sourcing diseases.
研究领域:
毒理学
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神经科学
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分子生物学
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表观遗传学
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发育生物学
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高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2024
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IF 12.2
Qing Wang,Yefei Huang,Min Jiang,Yu Tang,Qinzhi Wang,Longlong Bai,Chenglong Yu,Xinyue Yang,Kun Ding,Weimin Wang,Jin Bai,Yansu Chen
N-Nitroso compounds (NOCs) are recognized as important factors that promote gastric cancer development, but the specific effects and potential mechanisms by which NOC exposure promotes gastric cancer are still poorly understood. In this study, we explored the effects and potential molecular mechanisms of NOCs on the promotion of gastric cancer using methylnitronitrosoguanidine (MNNG), a classical direct carcinogen of NOC. The results of in vivo and in vitro experiments showed that chronic and low-concentration MNNG exposure significantly promoted the malignant progression of tumors, including cell migration, cell invasion, vasculogenic mimicry (VM) formation, cell spheroid formation, stem cell-like marker expression, and gastric cancer growth and metastasis. Mechanistically, we revealed that demethylase ALKBH5 regulated the level of the N6‑methyladenosine (m6A) modification in the 3′UTR and CDS region of the ZKSCAN3 mRNA to promote ZKSCAN3 expression, mediated the binding of ZKSCAN3 to the VEGFA promoter region to regulate VEGFA transcription, and participated in MNNG-induced gastric cancer cell migration, invasion, VM formation, cell spheroid formation, stem cell-like marker expression and ultimately gastric cancer progression. In addition, our study revealed that ALKBH5-ZKSCAN3-VEGFA signaling was significantly activated during MNNG-induced gastric carcinogenesis, and further studies in gastric cancer patients showed that ALKBH5, ZKSCAN3, and VEGFA expression were upregulated in cancers compared with paired gastric mucosal tissues, that ALKBH5, ZKSCAN3, and VEGFA could serve as important biomarkers for determining patient prognosis, and that the molecular combination showed greater prognostic value. These findings provide a theoretical basis for developing gastric cancer interventions for NOCs and for determining gastric cancer progression.
研究领域:
肿瘤学
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分子生物学
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癌症生物学
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表观遗传学
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高影响因子
Nature Communications
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2024
·Vol. 15
·期 1
·1-16
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IF 14.7
Zhuang Tao,Chen Mei-Hua,Wu Ruo-Xi,Wang Jing,Hu Xi-De,Meng Ting,Wu Ai-Hua,Li Yan,Yang Yong-Feng,Lei Yu,Hu Dong-Hua,Li Yan-Xiu,Zhang Li,Sun Ai-Jun,Lu Wei,Zhang Guan-Nan,Zuo Jun-Li,Ruan Cheng-Chao
Cardiac macrophage contributes to the development of cardiac fibrosis, but factors that regulate cardiac macrophages transition and activation during this process remains elusive. Here we show, by single-cell transcriptomics, lineage tracing and parabiosis, that cardiac macrophages from circulating monocytes preferentially commit to macrophage-to-myofibroblast transition (MMT) under angiotensin II (Ang II)-induced hypertension, with accompanying increased expression of the RNA N6-methyladenosine demethylases, ALKBH5. Meanwhile, macrophage-specific knockout of ALKBH5 inhibits Ang II-induced MMT, and subsequently ameliorates cardiac fibrosis and dysfunction. Mechanistically, RNA immunoprecipitation sequencing identifies interlukin-11 (IL-11) mRNA as a target for ALKBH5-mediated m6A demethylation, leading to increased IL-11 mRNA stability and protein levels. By contrast, overexpression of IL11 in circulating macrophages reverses the phenotype in ALKBH5-deficient mice and macrophage. Lastly, targeted delivery of ALKBH5 or IL-11 receptor α (IL11RA1) siRNA to monocytes/macrophages attenuates MMT and cardiac fibrosis under hypertensive stress. Our results thus suggest that the ALKBH5/IL-11/IL11RA1/MMT axis alters cardiac macrophage and contributes to hypertensive cardiac fibrosis and dysfunction in mice, and thereby identify potential targets for cardiac fibrosis therapy in patients.
研究领域:
心脏病学
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分子生物学
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免疫学
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高影响因子
Journal of Translational Medicine
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2025
·Vol. 23
·期 1
·1271
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IF 7.5
Zhu Jiru,Xia Yutong,Li Xiang,Wen Yingying,Zhang Xuhong,Jin Le,Zhang Liyue,Wang Ziyu,Li Yanqing,Yu Xin,Chen Zhitong,Ge Jiayun,Shen Ye,Tong Jianping
Background Myopia has reached epidemic proportions globally with limited therapeutic targets. This study elucidated the pivotal role of retinal epitranscriptomic regulation in myopia pathogenesis, aiming to identify novel therapeutic targets for myopia control. Methods The study enrolled 38 myopic patients scheduled for refractive surgery to analyze peripheral blood N6-methyladenosine (m6A) levels. In parallel, we established a murine form-deprivation myopia (FDM) model to examine retinal m6A dynamics and Alkylation repair homolog protein 5 (ALKBH5) expression. Pharmacological interventions using the ALKBH5 inhibitor DDO-2728 and Extracellular signal-related kinases 1 and 2 (ERK1/2) activator Ro67-7476 were administered intravitreally. To genetically validate the role of ALKBH5, ALKBH5-knockout (ALKBH5-KO) mice were subjected to the FDM model. Comprehensive assessments included refractive measurements, retinal structure analysis, and molecular profiling of inflammatory and extracellular matrix (ECM) markers. Statistical analyses employed Pearson’s correlation for clinical data and ANOVA with Tukey’s post hoc tests for experimental comparisons. Results Clinical data revealed significant inverse correlations between blood m6A levels and myopia severity (refraction: r = 0.435, p < 0.01; axial length: r = -0.338, p < 0.05). FDM retinas exhibited progressive m6A reduction and specific upregulation of ALKBH5 in the ganglion cell layer (GCL), with no changes observed in other m6A regulators. Mechanistic studies demonstrated that intravitreal injection of the selective ALKBH5 inhibitor DDO-2728 (2.5 mM) effectively attenuated myopia progression, mitigated retinal thinning, suppressed inflammatory mediators, and modulated early ECM dysregulation through specific suppression of ERK1/2 hyperphosphorylation. Crucially, genetic ablation of ALKBH5 recapitulated the protective phenotype of the inhibitor, conferring significant resistance to myopia development and associated retinal pathology. Importantly, co-administration of the ERK1/2 activator Ro 67-7476 completely abolished DDO-2728’s protective effects. Conclusions Peripheral blood m6A reduction correlates with human myopia severity. Retinal ALKBH5 upregulation drives myopia progression through ERK1/2 hyperactivation. Both pharmacological inhibition and genetic deletion of ALKBH5 effectively protect against myopia, attenuate inflammatory responses, and mitigate ECM dysregulation, mechanistically linked to the suppression of ERK1/2 signaling. These findings collectively identify ALKBH5 as a novel therapeutic target and DDO-2728 as a promising strategy for myopia control. Graphical
研究领域:
眼科学
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分子生物学
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表观遗传学
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信号转导
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药理学
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高影响因子
Nature Communications
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2026
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IF 18.1
Yang Yong-Feng,Li Yan,Chen Mei-Hua,Hu Xi-De,Hua Yi-Jie,Shi Xiao-Dong,Hu Dong-Hua,Meng Ting,Sheng Cheng-Yu,Zhuang Tao,Ruan Cheng-Chao,Zuo Xiang-Rong,Lei Yu,Li Ruo-Gu
Interactions between nervous system and adipose tissue are involved in the regulation of adipocyte browning. However, whether microglial activation participates in the regulation of cold exposure-induced sympathetic innervation and adipocyte browning is unclear. Here, we demonstrate that RNA m6A demethylase ALKBH5 in the microglia nucleus of hypothalamus (PVH) region inhibits cold exposure-induced adipocyte browning in the subcutaneous white adipose tissue (sWAT) of male mice. The single nuclear-RNA sequencing, brain slice electrophysiological assay and pathological analyses show that microglia-specific ALKBH5 knockout (MCKO) enhances sympathetic activation and adipocyte browning in the sWAT during cold exposure. Mechanistically, MCKO enhances m6A methylation of Ntf5 mRNA and upregulates Ntf5 expression in the microglia. Ntf5 then activates PVH-TrkB neurons and downstream sympathetic innervation in the sWAT. During aging, MCKO prevents brown adipocyte decline in the sWAT, and promotes metabolic activity. These findings fill a gap in the research of microglia in neuron-mediated adipocyte browning, and suggest that microglia may be a potential therapeutic target for the treatment of individuals with metabolic disorders.
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BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE
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2026
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IF 5
Guihong Li,Ting Lei,Erpeng Liu,Jing Shen,Dun Liu,Xuqiang Zhu
USP53 upregulation is identified as an independent risk factor for GBM progression. • USP53 upregulation is associated with poor prognosis of GBM patients. • USP53 promotes cell growth and the tumorigenicity of GBM cells. • USP53 removes ubiquitination from ALKBH5 and enhances its stability. • ALKBH5 removes the m6A modification of TERT and increases its RNA stability. Glioblastoma (GBM) is the most common malignant glioma, with a high recurrence rate and a poor prognosis. Deubiquitinating enzyme ubiquitin-specific peptidase 53 (USP53), known to enhance the stability of downstream proteins, exhibits distinct functions in different tumors. The function of USP53 in GBM progression remains unclear. Analysis of GSE104267 and GSE4290 datasets revealed significant USP53 upregulation in GBM tissues, a finding confirmed by TCGA data comparing GBM ( n = 163) with normal brain tissues ( n = 207). In univariate and multivariate cox regression analysis, USP53 was identified as an independent risk factor (univariate, hazard ratio (HR) =1.5357, p = 0.0229; multivariate, HR =1.4851, p = 0.0385) for GBM progression. Patients with high expression of USP53 had worse prognosis compared to those with low expression of USP53. Functionally, USP53 knockdown potently inhibited proliferation and induced apoptosis in LN229 and A172 cells in vitro and attenuated tumorigenicity of LN229 cells in vivo . Conversely, forced expression of USP53 had a carcinogenic effect. Mechanistically, USP53 targeted ALKB homolog 5 (ALKBH5) for deubiquitination, thereby stabilizing the protein and prolonging its half-life. Restoring ALKBH5 expression rescued the proliferative deficit in USP53-silenced LN229 cells. Telomerase catalytic subunit telomerase reverse transcriptase (TERT) was identified as a downstream effector of the USP53/ALKBH5 axis, where ALKBH5 upregulated TERT RNA expression by erasing m6A modifications at position 1989 bp on TERT mRNA. Collectively, those observations demonstrate that USP53/ALKBH5 axis drives GBM progression by targeting TERT, indicating that USP53/ALKBH5 axis may serve as a potential therapeutic target for GBM. Download: Download high-res image (229KB) Download: Download full-size image
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Cell Death & Disease
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2025
·Vol. 16
·期 1
·1-15
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IF 9.6
Cen Junjie,Zhao Dongliang,Shi Xin,Chen Jinlong,Zhou Hang,Liang Yanping,Gui Chengpeng,Chen Wei,Luo Junhang,Chen Xu
Renal cell carcinoma (RCC) remains a significant clinical challenge due to its increasing incidence and resistance to conventional therapy. This study identifies an unprecedented mechanism by which mesencephalic astrocyte-derived neurotrophic factor (MANF) contributes to RCC progression. From single-cell transcriptome, we observed a significant upregulation of MANF in RCC compared to its progenitor cells, the proximal tubular cells. Mechanistically, decreased N6-methyladenosine (m6A) modification on MANF mRNA, mediated by the upregulation of the m6A demethylase ALKBH5, led to reduced RNA degradation and increased MANF protein expression. Furthermore, while the common von Hippel-Lindau (VHL) deletion in RCC results in the accumulation of misfolded proteins and endoplasmic reticulum (ER) stress, increased MANF efficiently mitigated ER stress by binding to phosphorylated inositol-requiring enzyme-1 alpha (IRE1α) and inhibited its phosphorylation, indicating that MANF-mediated ER stress resistance compensates for the negative effects of VHL depletion and contributes to the survival of RCC. Our findings highlight a potential therapeutic strategy for RCC by targeting the m6A-mediated regulation, directly degrading MANF protein or inhibiting its function.
研究领域:
分子生物学
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癌症研究
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生物化学
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细胞生物学
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转化医学
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高影响因子
Translational Research
·
2026
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IF 6.1
Liying Luo,Lei Wang,Dan Ma,Wenxiu Liu,Zhiyu Shi,Chunnan Liu,Meng Zhao,Yufeng Bai,Jiukun Lu,Jinyu Chi
Hypertension is a chronic disease accompanied by vascular remodeling. As an m6A demethylase, ALKBH5 is implicated in cardiovascular diseases. Herein, we demonstrated that ALKBH5 expression was upregulated in the aortic tissues of spontaneously hypertensive rats (SHR), with predominant localization in vascular smooth muscle cells (VSMCs). To define ALKBH5 function, we generated an SM22α-driven, VSMC-specific Alkbh5 -knockdown AAV9 vector and administered a single tail-vein injection (11 × 10¹² v.g./rat) to 4-week-old male SHRs. Alkbh5 knockdown markedly lowered arterial pressure in SHRs, ameliorated left ventricular remodeling, attenuated aortic media thickening and extracellular matrix deposition, and suppressed the contractile-to-synthetic/proliferative phenotypic switch of VSMCs. Primary thoracic aortic VSMCs were isolated from 16-week-old SHRs and Wistar-Kyoto (WKY) rats, with Alkbh5 knockdown achieved via adenoviral transduction. Separately, primary human aortic smooth muscle cells were pretreated with 1 μM angiotensin II for 24 h prior to adenoviral infection. In vitro assays further verified that ALKBH5 knockdown suppressed proliferation and the contractile-to-synthetic/proliferative phenotypic switch in VSMCs in hypertensive context. Subsequent mechanistic studies revealed that ALKBH5 knockdown enhanced the stability of TEAD3 mRNA by increasing its m6A modification; the upregulated TEAD3 could directly bind to the PDZK1 promoter and promote its transcription. Moreover, PDZK1 knockdown reversed the inhibitory effect of ALKBH5 knockdown on the abnormal proliferation of VSMCs. In conclusion, ALKBH5 knockdown inhibits the abnormal phenotypic switch of VSMCs through the TEAD3/PDZK1 axis, thereby alleviating vascular remodeling caused by hypertension. These findings suggest that ALKBH5 may serve as a potential therapeutic target for hypertension.
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高影响因子
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