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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PubMed
高影响因子
AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY
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2023
·Vol. 90
·期 5
·e13791
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IF 3.6
Wenzhi Wang,Jianyong Shi,Lei Zheng
The emerging role of microRNA-935 (miR-935) in modulating cancer progression has been recognized. However, its role in regulating choriocarcinoma (CCA) development and progression remains unknown. The present work aims to reveal the effect of miR-935 on CCA cell tumor properties and the related mechanism. The RNA expression of methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit (METTL3), miR-935, and gap junction protein alpha 1 (GJA1) was detected by quantitative real-time polymerase chain reaction. Protein expression of GJA1, Ki67, and METTL3 was measured by western blotting and immunohistochemistry assays. CCK-8 and colony formation were used to analyze cell proliferation. Transwell assays were performed to assess cell migration and invasion. Angiogenesis was investigated by tube formation assay. Xenograft mouse model assay was used to determine miR-935-mediated effect on tumor formation in vivo. The luciferase reporter assay and RNA pull-down assay were used to verify the relationship between miR-935 and GJA1. MeRIP assay was used to analyze the m6A methylation of pri-miR-935. MiR-935 expression was significantly upregulated in CCA tissues and cells when compared with control groups. MiR-935 overexpression promoted CCA cell proliferation, migration, invasion, and tube formation and tumor tumorigenesis in vitro and in vivo, but miR-935 knockdown showed the opposite effects. In addition, miR-935 targeted GJA1 and mediated CCA cell tumor properties by negatively regulating GJA1 expression. METTL3 promoted miR-935 maturation by inducing m6A methylation of pri-miR-935, and its overexpression contributed to CCA cell tumor properties through the regulation of miR-935. METTL3 promoted choriocarcinoma progression by m6A-dependently activating the miR-935/GJA1 pathway.
研究领域:
癌症生物学
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分子生物学
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细胞生物学
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表观遗传学
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PubMed
高影响因子
ENVIRONMENTAL POLLUTION
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2020
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IF 6.793
Tian-Xin Zhao,Jun-Ke Wang,Lian-Ju Shen,Chun-Lan Long,Bin Liu,Yi Wei,Lin-Dong Han,Yue-Xin Wei,Sheng-De Wu,Guang-Hui Wei
Di-(2-ethylhexyl) phthalate (DEHP) is a common environmental endocrine disrupting chemical that may induce male reproductive disorders. Exposure to DEHP at a prepubertal stage could lead to prepubertal testicular injury, but the underlying mechanisms remain unclear. In this study, we exposed Sprague-Dawley rats to 0, 250, and 500 mg DEHP per kg body weight per day at the prepuberty stage from postnatal day 22 (PND 22) to PND 35 by oral gavage. Testicular injury and oxidative stress were evaluated, and the levels of 6-methyladenosine (m6A) modification and expression of modulator genes for RNA methylation were measured in testes . Furthermore, m6A modification of the important antioxidant transcription factor Nrf2 was analyzed using methylated RNA immunoprecipitation qPCR. Our results show that DEHP worsened testicular histology, decreased testosterone concentrations, downregulated expression of spermatogenesis inducers, enhanced oxidative stress, inhibited the Nrf2-mediated antioxidant pathway, and increased apoptosis in testes. Additionally, DEHP increased global levels of m6A RNA modification and altered the expression of two important RNA methylation modulator genes, FTO and YTHDC2 . Moreover, m6A modification of Nrf2 mRNA increased upon DEHP exposure. Overall, these findings link oxidative stress imbalance with epigenetic effects of DEHP toxicity and provide insight into the testicular toxicity of DEHP from the new perspective of m6A modification.
研究领域:
毒理学
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生殖生物学
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表观遗传学
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PubMed
高影响因子
Discover Oncology
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2024
·Vol. 15
·期 1
·1-18
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IF 2.8
Feng Rui,Li Haolin,Meng Tong,Fei Mingtian,Yang Cheng
Purpose This study aimed to construct an m6A and cuproptosis-related long non-coding RNAs (lncRNAs) signature to accurately predict the prognosis of kidney clear cell carcinoma (KIRC) patients using the information acquired from The Cancer Genome Atlas (TCGA) database. Methods First, the co-expression analysis was performed to identify lncRNAs linked with N6-methyladenosine (m6A) and cuproptosis in ccRCC. Then, a model encompassing four candidate lncRNAs was constructed via univariate, least absolute shrinkage together with selection operator (LASSO), and multivariate regression analyses. Furthermore, Kaplan–Meier, principal component, functional enrichment annotation, and nomogram analyses were performed to develop a risk model that could effectively assess medical outcomes for ccRCC cases. Moreover, the cellular function of NFE4 in Caki-1/OS-RC-2 cultures was elucidated through CCK-8/EdU assessments and Transwell experiments. Dataset outcomes indicated that NFE4 can have possible implications in m6A and cuproptosis, and may promote ccRCC progression. Results We constructed a panel of m6A and cuproptosis-related lncRNAs to construct a prognostic prediction model. The Kaplan–Meier and ROC curves showed that the feature had acceptable predictive validity in the TCGA training, test, and complete groups. Furthermore, the m6A and cuproptosis-related lncRNA model indicated higher diagnostic efficiency than other clinical features. Moreover, the NFE4 function analysis indicated a gene associated with m6A and cuproptosis-related lncRNAs in ccRCC. It was also revealed that the proliferation and migration of Caki-1 /OS-RC-2 cells were inhibited in the NFE4 knockdown group. Conclusion Overall, this study indicated that NFE4 and our constructed risk signature could predict outcomes and have potential clinical value.
研究领域:
生物信息学
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癌症生物学
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分子生物学
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基因组学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2023
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IF 6.2
Huahui Gao,Xinmei Song,Qirong Chen,Ruo Yuan,Yun Xiang
The demethylase of fat mass and obesity related protein (FTO) is critical to regulate the dynamic N 6 -methyladenosine (m6A) modification of eukaryotic mRNAs, and its overexpression has found to be closely related to the initiation of several cancers. On the basis of a target-promoted specific activation of DNAzyme strategy coupled with self-primer exponential amplification reaction (SPEXPAR) cycles and DNA supersandwich assemblies, the highly sensitive and label-free electrochemical FTO assay approach is established. The modification of the catalytic core nucleobase of the DNAzyme probe by m6A can inhibit its cleavage activity. The presence of target FTO catalyzes the elimination of the methyl group to restore the DNAzyme activity, which cleaves the hairpin substrates to trigger the SPEXPAR for yielding many ssDNAs. The capture of these DNAs on the sensor electrode leads to the initiation of supersandwich assembly formation of long dsDNAs . Tremendous electrochemical signal probe of [Ru(NH 3 ) 6 ]Cl 3 are then absorbed on these dsDNAs to produce highly amplified catalytic currents with the assistance of K 3 [Fe(CN) 6 ] for detecting trace FTO with 63.1 fM detection limit. Furthermore, the sensor can be employed for selective assay of FTO in cell lysates, revealing the great potential of this sensing strategy for biomedical and biological study applications.
研究领域:
生物化学
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分子生物学
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分析化学
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生物医学工程
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2023
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IF 8.4
Xixiang Xie,Xiyu Liu,Wenxi Lu,Tao Wu,Jian He,Zhiming Deng,Faquan Lin,Pan Wu,Bing Wang,Yong Huang
N 6 -methyladenosine (m 6 A) modification has an impact on RNA metabolism, and it has been demonstrated that m 6 A regulatory mechanisms play an important role in tumorigenesis and may serve as a potential hepatocellular carcinoma (HCC) marker. We constructed a dual-signal amplification strategy electrochemical biosensor based on biocatalytic silver deposition and hybridization chain reaction (HCR). Enzymatic silver deposition was performed on the electrode and detected by linear sweep voltammetry (LSV), which was used for the quantitative detection of m 6 A. There was a nice linear relationship between the current response values and the logarithm of the m 6 A RNA concentration in the range of 0.5 pM-5 μM, with a detection limit as low as 25 fM. The concentration of m 6 A detected by the sensor constructed in this work is consistent with the results obtained using commercial colorimetric immunoassays . What's more, the analysis of m 6 A RNA in the serum of HCC patients using the developed sensor showed the high analytical efficiency of the designed strategy, which was superior to three other markers (AFP, CEA, CA199). The constructed electrochemical biosensor has good sensitivity, specificity and reproducibility for the detection of m 6 A RNA, and the sensing platform has promising clinical applications in the diagnosis and prognosis of HCC.
研究领域:
癌症生物学
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分子生物学
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电化学
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生物技术
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临床诊断
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高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
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2022
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IF 10.753
Haowei Fu,Xiaofang Zhu,Qiannan Di,Jiahui Sun,Qianqian Jiang,Qian Xu
Di(2-ethyl-hexyl) phthalate (DEHP), an environmental endocrine disruptor, can destroy the sperm genomic integrity and impairs spermatogenesis. N6-methyladenosine (m6A) is involved in the cellular effects of DEHP. However, the genotoxic effect of DEHP on spermatocytes and the possible role of m6A in this process remain unclear. This study demonstrated that m6A alleviates DEHP genotoxicity in GC-2 cells. In DEHP-treated mice, DNA double-strand breaks (DSBs) were induced in the testis and spermatocytes. To further explore the molecular mechanism of DEHP genotoxicity on spermatocytes, GC-2 cells were exposed to DEHP. DEHP produced distinct genotoxicity and caused DSBs, which led to the inhibition of DNA synthesis and cell cycle arrest. The DNA damage response (DDR) was initiated to repair the DSBs induced by environmentally relevant levels of DEHP (100 μM and 200 μM). During this process, METTL3 upregulated m6A, which facilitated the DDR via stabilizing the DNA damage repair factors ( Rad51 and Xrcc5 ) mRNA to maintain the pro-survival state. Moreover, Mettl3 knockdown partially inhibited DDR. Interestingly, high-dose DEHP (400 μM and 600 μM) directly induced apoptosis rather than the pro-survival state. Altogether: METTL3-mediated m6A participates in maintaining the pro-survival state by upregulating DDR, providing guidance for mitigating the genotoxicity of environment-related level DEHP exposure.
研究领域:
毒理学
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生殖生物学
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分子生物学
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环境科学
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PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
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2024
·Vol. 22
·期 1
·1-21
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IF 10.6
Yu Jiang,Gao Yidan,Liu Fei,Zhang Yuyu,Li Jianda,Ding Luogang,Ren Sufang,Yang Jie,Jiao Jian,Feng Gong,Chen Zhi,Sun Wenbo,Wu Jiaqiang
Background Animal and human health are seriously threatened by bacterial infections, which can lead to bacteremia and extremely high rates of morbidity and mortality. Recently, there have been reports indicating the involvement of exosomal circular RNAs (circRNAs) in a range of human disorders and tumor types. However, the role of exosomal circRNAs in bacterial infection remains elusive. Methods We extracted and identified exosomes from the culture medium of PIEC cells infected with or without Glaesserella parasuis . RNA sequencing analysis was performed on the exosomes to screen and identify circRNAs (circHIF1α) associated with Glaesserella parasuis infection. PIEC cells were infected with Staphylococcus aureus or Streptococcus suis 2 to further determine whether exosome-derived circHIF1α was the crucial circHIF1α associated with bacterial infections. The transmission process of exosomes and their circHIF1α between cells was clarified via exosome tracing and co-culture assay. Moreover, the mechanism of circHIF1α being packaged into exosomes was explored, and the effects of exosomes and their circHIF1α on cell proliferation, DNA damage and cell cycle were analyzed. In addition, the binding mode and site of interacting proteins with circHIF1α were further determined. In vivo and in vitro, the role of exosomes and their circHIF1α in host resistance to bacterial infection was confirmed. Results We first discovered a new circHIF1α that was very stable and detectable, encapsulated into exosomes by hnRNPA2B1, and whose expression in exosomes of bacterially infected PIEC cells significantly decreased. Additionally, exosomal circHIF1α reduced bacterial infection both in vitro and in vivo and suppressed the growth of reception cells. Mechanistically, the circHIF1α interacted with the KH domain of IGF2BP3 in an m6A-modified manner, which mediated DNA damage to arrest the cells at the G1/S phase through the interaction between the regulator of Chromosome Condensation 2 (RCC2) and γ-H2AX protein. Exosomal circHIF1α is a unique therapeutic target for bacterial infection since this work highlights its critical function in fighting bacterial infection.
研究领域:
分子生物学
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微生物学
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细胞生物学
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传染病学
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PubMed
高影响因子
Science China-Life Sciences
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2025
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IF 9.5
Mi Yujie,Huang Chenyang,Liu Donghong,Li Tengwei,Ying Xiner,Cao Jie,Liu Jianzhao
As one of the most prevalent messenger RNA modifications in eukaryotes, N 6 -methyladenosine (m 6 A) regulates various physiological and pathological processes. The RNA m 6 A sequencing techniques have significantly advanced the understanding of m 6 A biology, however, there is a lack of tools to in situ spatially characterize RNA m 6 A and its interacting components inside cells. Spatial information could significantly promote our understanding of the m 6 A dynamics and functions in complex biological contexts. While traditional m 6 A antibody immunostaining method images global cellular m 6 A sites, we here develop a non-antibody-based fluorescent imaging tool to in situ visualize single m 6 A site of specific cellular RNA, named m 6 A REading and Fluorescent Imaging In situ (m 6 A-REFII), which contains m 6 A reader module, hybridization probe module for targeting specific m 6 A-locating RNA sequence, and fluorescent signaling module. We successfully leveraged m 6 A-REFII to visualize m 6 A-modified RNAs at distinct subcellular localizations as well as to reveal the dynamic changes of targeted m 6 A sites in response to CRISPR-mediated m 6 A reduction and heat shock stimuli. Furthermore, we demonstrated the compatibility of m 6 A-REFII with protein immunofluorescence staining and validated the finding that the scaffold attachment factor B protein binds m 6 A-modified long interspersed element-1 RNA. Collectively, m 6 A-REFII is a new tool empowering visualization of single m 6 A modification on specific RNAs at the single-cell level.
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高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2025
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IF 6.1
Jingyu Wang,Xiaoli Guo,Yujiao Chen,Wei Zhang,Jing Ren,Ai Gao
Chronic benzene exposure causes hematotoxicity, yet the underlying molecular mechanisms remain incompletely understood. N6-methyladenosine (m6A) RNA modification has emerged as a critical regulator in various diseases, but its role in benzene-induced hematotoxicity is unclear. In this study, m6A mRNA microarray analysis revealed widespread alterations in m6A methylation following benzene exposure. Gene Ontology (GO) enrichment identified ubiquitin conjugating enzyme E2 G2 (UBE2G2) as a key m6A-methylated transcript associated with protein degradation pathways. Bioinformatics prediction and experimental validation (RIP-qPCR and MeRIP-qPCR) confirmed that the m6A reader YTHDF3 directly binds to UBE2G2 mRNA in an m6A-dependent manner. Benzene exposure downregulated YTHDF3 expression, leading to decreased UBE2G2 mRNA stability and expression. Overexpression of YTHDF3 reversed this suppression, confirming the regulatory relationship. Functionally, reduced UBE2G2 impaired ubiquitination of acyl-CoA synthetase long-chain family member 4 (ACSL4), resulting in its stabilization and upregulation, which promoted lipid peroxidation and ferroptosis. Notably, melatonin treatment ameliorated benzene-induced hematotoxicity by restoring the YTHDF3/UBE2G2/ACSL4 axis. In summary, this study identifies a novel epigenetic pathway—YTHDF3/m6A/UBE2G2/ACSL4—that mediates benzene-induced ferroptosis, and demonstrates the protective effect of melatonin through modulation of this axis.
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高影响因子
BIOCHEMICAL PHARMACOLOGY
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2025
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IF 5.6
Jing Zhang,Xuewei Yang,Xing Zhao,Gang Luo,Hong Jiang
Renal fibrosis is recognized as a critical pathway leading to end-stage renal disease in nearly all cases of chronic kidney disease (CKD). Analysis of the gene expression omnibus (GEO) database revealed that the transcription factor Krüppel-like factor 3 (KLF3) was elevated in kidney tissues of CKD patients, however, its specific role in CKD-associated renal fibrosis is yet to be deciphered. This study aimed to investigate the function of KLF3 on unilateral ureteral obstruction (UUO) mice and transforming growth factor-β1 (TGF-β1)-stimulated HK2 cells. Our data demonstrated that KLF3 was remarkably upregulated in the renal cortex of UUO mice and TGF-β1-treated HK-2 cells. We further confirmed that the upregulation of KLF3 was associated with its N6-methyladenosine (m6A) methylation modification. Knockdown of METTL14 reduced m6A methylation level of KLF3 and inhibited its mRNA stability. In vivo results suggested that KLF3 knockdown alleviated the pathological changes in renal tissues triggered by UUO. Additionally, KLF3 silencing mitigated renal fibrosis both in vivo and in vitro , as accompanied by decreased expression of Collagen I, Vimentin, α-SMA and Fibronectin. Mechanistically, KLF3 transcription activated dimethylarginine dimethylaminohydroxylase 2 (DDAH2) and upregulated its expression. Rescue experiments substantiated that DDAH2 overexpression eliminated the effect of KLF3 knockdown on renal fibrosis. Collectively, our findings suggest that inhibition of KLF3 may serve as a potential strategy to prevent CKD-associated renal fibrosis.
研究领域:
分子生物学
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肾脏病学
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PubMed
高影响因子
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY
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2025
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IF 4.7
Long Li,Quanyou Chai,Chunling Guo,Junyi Wei,Yuqiao Qin,Huimin Liu,Zhaoyang Lu
Aim Vascular calcification (VC) is a major adverse cardiovascular event in chronic kidney disease (CKD) patients. N6-methyladenosine (m6A) modification is vital for many biological processes, but its function and possible molecular mechanisms in VC are poorly understood. This study aimed to clarify the function and molecular mechanisms of N6-adenosine-methyltransferase-like 3 (METTL3) in VC. Methods and results The results of the bioinformatic analysis showed that METTL3 expression was significantly upregulated in calcified VSMCs. This finding was corroborated by phosphate-induced VSMCs calcification models and 5/6 nephrectomy-induced CKD mouse VC models. Afterward, Alizarin Red S staining and m6A dot blot analysis demonstrated METTL3 overexpression elevated m6A levels and encouraged calcification in VSMCs and mouse aortic rings, while METTL3 knockdown decreased m6A levels and inhibited calcium deposition in these experimental models. Furthermore, METTL3 promoted VC via the PTEN/AKT pathway, and MeRIP verified that METTL3 induced PTEN mRNA degradation by modifying it with m6A. In addition, molecular docking simulations and DARTS assays revealed that quercetin is a natural small-molecule inhibitor of METTL3. The current investigation demonstrated that quercetin mitigated VC by reducing METTL3-dependent m6A levels in vivo and in vitro. Conclusion In conclusion, this study unraveled the pathogenic mechanism of METTL3-mediated m6A modification in VC and provided new insights to establish METTL3 as a therapeutic target for VC.
研究领域:
分子生物学
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心血管疾病
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表观遗传学
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PubMed
高影响因子
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