Journal of Translational Medicine
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2024
·Vol. 22
·期 1
·1-19
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IF 6.1
Wang Runze,Xue Wei,Kan Feifei,Zhang Huiying,Wang Di,Wang Lei,Wang Jianwen
Background Diabetic retinopathy (DR) is the leading cause of blinding eye disease among working adults and is primarily attributed to the excessive proliferation of microvessels, which leads to vitreous hemorrhage and retinal traction, thereby significantly impairing patient vision. NSUN2-mediated RNA m 5 C methylation is implicated in various diseases, and in this investigation, we focused on elucidating the impact of NSUN2 on the regulation of the expression of the downstream gene MUC1, specifically through RNA m 5 C methylation, on the progression of DR. Method Utilizing Microarray analysis, we examined patient vitreous fluid to pinpoint potential therapeutic targets for DR. Differential expression of NSUN2 was validated through qRT-PCR, Western blot, and immunofluorescence in human tissue, animal tissue, and cell model of DR. The relationship between NSUN2 and DR was explored in vitro and in vivo through gene knockdown and overexpression. Various techniques, such as MeRIP-qPCR and dot blot, were applied to reveal the downstream targets and mechanism of action of NSUN2. Results The levels of both NSUN2 and RNA m 5 C methylation were significantly elevated in the DR model. Knockdown of NSUN2 mitigated DR lesion formation both in vitro and in vivo. Mechanistically, NSUN2 promoted MUC1 expression by binding to the RNA m 5 C reader ALYREF. Knockdown of ALYREF resulted in DR lesion alterations similar to those observed with NSUN2 knockdown. Moreover, MUC1 overexpression successfully reversed a series of DR alterations induced by NSUN2 silencing. Conclusions NSUN2 regulates the expression of MUC1 through ALYREF-mediated RNA m 5 C methylation, thereby regulating the progression of DR and providing a new option for the treatment of DR in the future.
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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高影响因子
ANALYTICA CHIMICA ACTA
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2025
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IF 6
Mimi Li,Lina Wang,Yue Hu,Yi Liu,Shuang Xu,Kexing Peng,Chenghong Li,Hui Huang,Lichao Fang,Lulu Li,Huamin Liu,Xiaolong Wang,Junsong Zheng
Sensitive and efficient detection of RNA methylation sites is considered an integral part of epigenetic assessment. Contrary to previous studies on the detection of RNA m5C or m6A methylation alone, this study innovatively designed a biosensor capable of detecting RNA m5C or m6A methylation at the same time. Substances with specific electrochemical reactions such as ferrocene (FC) or methylene blue (MB) were modified onto the hairpin probe. When the hairpin probe was activated under specific conditions, it triggered a dendritic nonlinear hybridization chain reaction (HCR), which resulted in signal amplification. Gold (Au) and iron tetraoxide (Fe 3 O 4 ) composite nanomaterials were employed as the linking materials: the carboxylated ends of Fe 3 O 4 were connected to an antibody that specifically recognizes the m5C and m6A methylation sites, while Au nanoparticle ends adhering to the carboxylated Fe 3 O 4 surface are connected to the HCR enhanced signal amplifier. The m5C antibody was linked to Fc-containing HCR amplification products by this material, and similarly, the m6A antibody was linked to MB-containing amplifiers. Thus, the dandelion complex, a multifunctional body with methyl recognition and signal amplification, was formed. The capture probe immobilized on the surface of the gold electrode by Au–S recognized the target RNA sequence by base complementary pairing. Upon addition of the dandelion complex, due to its multi-functionality, the amplified signals were carried to the electrode by the antigen-antibody recognition mechanism, generating a current signal. The positions and heights of the current signal peaks enabled rapid determination of target RNA methyl modification sites and their abundance. The detection limits of this biosensor for RNA m5C and m6A were 4.68 × 10 −16 M and 1.10 × 10 −15 M, respectively, and the linear range was from 10 −15 mol/L to 10 −8 mol/L. The sensor developed in this study has the advantages of cost-effectiveness, ease of fabrication, and fast response time, and has the potential to be promoted for RNA methylation detection.
研究领域:
分子生物学
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生物传感技术
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表观遗传学
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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
Zhe Du,Qingping Liu,Mengruo Wang,Yifu Gao,Qi Li,Yizhe Yang,Tianyu Lu,Lei Bao,Yaxian Pang,Haijun Wang,Yujie Niu,Rong Zhang
Accumulating evidence indicates that maternal exposure to carbon black nanoparticles (CBNPs) during gestation can induce multiple system abnormalities in offspring, whereas its potential mechanism in respiratory disease is still largely unknown. In order to explore the effect of maternal exposure to CBNPs on offspring’s lung and latent pathogenesis, we respectively established in vivo model of pregnant rats exposed to CBNPs and ex vivo model of lung epithelial cells treated with pups’ serum of pregnant rats exposed to CBNPs. After maternal exposure to CBNPs, epithelial-mesenchymal transition (EMT) and fibrosis levels increased as a result of DDRGK1-mediated reticulophagy upregulated in offspring’s lung. DDRGK1 as FAM134B’s cargo bound with FAM134B to mediate reticulophagy. Transcription factor “SP1” positively regulated DDRGK1 gene expression by binding to its promoter. Furthermore, the upregulation of NSUN2 elevated m 5 C methylation of SP1 mRNA and the protein level of SP1 subsequently increased through Ybx1 recognizing and stabilizing m 5 C-methylated SP1 mRNA, followed by the increased levels of reticulophagy and fibrosis in lung epithelial cells treated with offspring’s serum of matrix exposed to CBNPs during gestation. In conclusion, NSUN2/Ybx1/m 5 C-SP1 axis promoted DDRGK1-mediated reticulophagy, which played an important role in EMT-induced fibrosis in offspring’s lung tissue after maternal exposure to CBNPs during gestation.
研究领域:
呼吸系统疾病
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纳米毒理学
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分子生物学
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表观遗传学
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DOI
PubMed
高影响因子
Genes
·
2021
·Vol. 12
·期 10
·1488
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IF 4.096
Jizhong Zhao,Hongmei Hu,Sheng Wang,Li Wang,Rui Wang
In living organisms, RNA regulates gene expression, cell migration, differentiation, and cell death. 5-Methylcytosine is a post-transcriptional RNA modification in a wide range of RNA species, including messenger RNAs. The addition of m5C to RNA cytosines is enabled by theNSUNenzyme family, a critical RNA methyltransferase. In this study, natural lysines modified with special groups were synthesized. Through two rounds of positive screening and one round of negative screening, we evaluated and identified theMbPylRS-tRNACUAunnatural lysine substitution system, which specifically recognizes lysine with a defined group. Moreover, non-natural lysine substitution at C271 ofNSUN2active site and the subsequent fluorescent labeling was realized through the click reaction. Then, the function of theNSUN2mutant and its upregulatedCDK1gene as well as its effect on cell proliferation were evaluated. Efficient labeling and regulation ofNSUN2was achieved, laying the basis for further studies on the function and regulatory mechanism of upregulated genes.Keywords:RNA modification;m5C;NSUN enzyme;regulation;labeling
研究领域:
分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
BREAST CANCER RESEARCH AND TREATMENT
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2025
·Vol. 57
·期 4
·1064
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IF 3
Jie Hu,Qilong Liu,Bi Feng,Yanling Lu,Kai Chen
Purpose Esophageal cancer (EC) is a rapidly progressing malignancy characterized by a low survival rate and limited treatment success, largely due to late-stage detection, frequent recurrence, and a high propensity for metastasis, despite ongoing advances in therapeutic strategies. While oxaliplatin (L-OHP) is a potent chemotherapeutic agent that induces apoptosis in EC cells, its effectiveness is significantly hindered by the development of resistance. Materials and Methods The assessment of gene and protein expression was conducted through a combination of quantitative real-time polymerase chain reaction, Western blot, and immunohistochemical staining. Cell viability was assessed using the cell counting kit-8 assay. The interactions among ALYREF, TBL1XR1, KMT2E, and APOC1 were investigated through RNA immunoprecipitation, chromatin immunoprecipitation (ChIP), ChIP-reChIP, RNA pulldown, and dual-luciferase assays. An in vivo mouse model of EC was established. Results Expression levels of both APOC1 and ALYREF were elevated in L-OHP–resistant EC tissues and cell lines, and their silencing enhanced sensitivity to L-OHP. TBL1XR1 and KMT2E synergistically upregulated APOC1 expression. Moreover, ALYREF recognized the 5-methylcytosine (m5C) sites on TBL1XR1 and KMT2E mRNAs, stabilizing these transcripts and promoting APOC1 expression. The regulatory role of these interactions was further validated in vivo . Conclusion This study demonstrated that ALYREF interacted with the m5C sites on TBL1XR1 and KMT2E mRNAs, enhancing their stability and leading to increased transcription of APOC1, which in turn contributed to L-OHP resistance in EC. These findings suggest that targeting APOC1 could be a promising strategy for overcoming L-OHP resistance in EC.
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Cancer & Metabolism
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2025
·Vol. 13
·期 1
·1-17
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IF 5.3
Li Hao,Gong Tingyue,Zhao Yongheng,Luo Yang,Tang Shuibin,Wang Tingfeng,Lin Haiping,Zhong Ming
Purpose Cancer cells rely on serine biosynthesis for growth, but its regulation in colorectal cancer (CRC) remains not well understood. This study identifies the m 5 C methyltransferase NSUN2 (NOP2/Sun domain family, member 2) as a key regulator of serine biosynthesis, revealing a novel mechanism driving CRC progression. Methods The expression and prognostic value of NSUN2 were evaluated using bioinformatics analyses and immunohistochemistry (IHC) assays. The effects of NSUN2 on cellular serine biosynthesis, intracellular reactive oxygen species (ROS) levels, and apoptosis levels were analyzed both in vitro and in vivo. Additionally, RNA sequencing, Methylated RNA Immunoprecipitation sequencing (MeRIP-seq), RNA immunoprecipitation (RIP), and RNA stability assays were utilized to screen and validate the association between NSUN2 and phosphoglycerate dehydrogenase (PHGDH). Results NSUN2 was found to be highly expressed in CRC and associated with poor patient survival. PHGDH, a direct downstream target of NSUN2, plays a crucial role in NSUN2-mediated serine biosynthesis. Furthermore, inhibition of NSUN2 significantly reduced the intracellular NADH/NAD + and NADPH/NADP + ratios, leading to an increase in ROS levels and apoptosis levels, thereby inhibiting CRC progression. Additionally, NSUN2 enhances PHGDH expression and mRNA stability by binding to the “reader” protein m 5 C-Aly/REF export factor (ALYREF). Conclusions This study identified a novel NSUN2/ALYREF/m 5 C-PHGDH axis might be promising therapeutic targets for CRC.
研究领域:
肿瘤学
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分子生物学
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癌症研究
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RNA生物学
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代谢调控
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PubMed
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FREE RADICAL BIOLOGY AND MEDICINE
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2025
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IF 8.2
Yunpeng Zhu,Lingjiang Li,Yunshan Guo,Renji Wang,Yiguang Hao,Xin He
The formation of neutrophil extracellular traps (NETs) at the injury site contributes to secondary damage in spinal cord injury (SCI). This study investigated the function of EDIL3 in spinal cord ischemia-reperfusion injury (SCIRI) and its connection to NET formation. We established a mouse SCIRI model, which revealed significant neutrophil infiltration and NET formation within the injured spinal cord at 48 hours post-reperfusion. Therapeutic disruption of NETs with DNase1 or EDIL3 overexpression ameliorated multiple aspects of SCIRI pathology, including tissue damage, blood-spinal cord barrier (BSCB) disruption, neuroinflammation, neuronal death, and motor deficits. Endothelial cell-specific knockout of EDIL3, however, intensified these injuries. Mechanistically, EDIL3 inhibited PMA-induced NET formation in vitro by attenuating NF-κB activation and protected the endothelial barrier from NET-mediated damage in co-culture assays. Moreover, NSUN2 promoted m5C methylation of EDIL3 mRNA, facilitating its YBX1-dependent stabilization. Crucially, the exacerbation of SCIRI following NSUN2 knockdown was abrogated by EDIL3 upregulation. Collectively, the NSUN2-EDIL3 axis represents a key endogenous pathway that limits BSCB disruption and neuronal death in SCIRI by restraining NET formation.
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Biomarker Research
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2026
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IF 14.6
Ruixin Wu,Licheng Zhang,Peter JihCheng Wong,Chunming Sun,Yuan Feng,Xiaoyu Wang,Minjie Fu,Jinsen Zhang,Fufang Qiu,Xi Chen,Zhen Fan,Minfeng Shu,Wei Hua
IDH mutant gliomas, driven by the oncometabolite 2-hydroxyglutarate (2-HG), are associated with profound neurological morbidity and premature mortality. To address the unmet therapeutic needs, we investigated the mechanistic interplay between MTHFD2-driven one-carbon metabolism and ferroptosis susceptibility in these tumors. Our findings revealed that MTHFD2 upregulation, mediated through loss of m5C modification in chromatin associated RNAs (caRNAs)-establishes a metabolic vulnerability to ferroptosis. Crucially, we identified TAF15 as a pivotal RNA-binding protein that orchestrates the spatial recruitment of TET2 by bridging NSUN5-mediated RNA m5C methylation. Therapeutically, combinatorial targeting of MTHFD2 with its selective inhibitor and the hypomethylating agent decitabine induced ferroptosis in patient-derived IDH-mut glioma organoids, demonstrating potent ferroptosis activation. This work delineates an RNA epitranscriptomic-metabolism axis in glioma pathogenesis and provides a translational roadmap for exploiting metabolic dependencies in IDH-driven malignancies. CLINICAL TRIAL NUMBER: Not applicable.
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高影响因子
CELLULAR AND MOLECULAR LIFE SCIENCES
·
2026
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IF 6.5
Siqi Nian,Lulu Zhao,Xiaokang He,Ping Xia,Yanqing Liu,Baotong Hua,Jun Li,Hongxing Zhang
Background: Patients with heart failure (HF) frequently develop atrial fibrillation, but the precise molecular mechanisms are unclear, leading to limited therapeutic strategies. Methods: Isoproterenol hydrochloride (ISO)-induced mouse HF model and ISO-treated HL-1 cardiomyocyte model were employed. A combination of qRT-PCR, Western blot, ChIP, dual-luciferase reporter assay, MeRIP-qPCR, RIP-qPCR, actinomycin D assay, and mitochondrial function assessments were used to systematically investigate the regulatory mechanism of the ALYREF/MZF1/HTRA1 axis and its role in cardiac remodeling and atrial fibrillation susceptibility. Results: HTRA1 expression was elevated in HF mice and human atrial fibrillation data. In cellular models, HTRA1 overexpression induced mitochondrial damage, cellular hypertrophy, and inflammation via the HIF-1α pathway. The transcription factor MZF1 directly bound to and activated the HTRA1 promoter. NSUN2-mediated m5C modification of MZF1 mRNA was required for its recognition and stabilization by the m5C reader protein ALYREF, thereby positively regulating the MZF1/HTRA1 axis. In the ISO-induced HF mouse model, knockdown of ALYREF effectively reduced MZF1 and HTRA1 expression, alleviated myocardial hypertrophy, fibrosis, cardiac dysfunction, and atrial fibrillation inducibility. Conclusion: This study elucidates a novel signaling axis wherein NSUN2-mediated m5C modification of MZF1 mRNA enables its stabilization by ALYREF, leading to transcriptional upregulation of HTRA1. This axis drives cardiomyocyte mitochondrial dysfunction, hypertrophy, and inflammation in the context of HF, ultimately increasing susceptibility to atrial fibrillation.
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高影响因子
Scientific Reports
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2025
·Vol. 15
·期 1
·1-11
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IF 3.9
Xu Guobin,Wang Junlong,Gu Na,Yang Tiansong
Pyroptosis is a pathogenenic mechanism of polycystic ovary syndrome (PCOS). N5-methylcytosine (m5C) methylation is a RNA modification that affects the progression of diseases. However, its role in PCOS remains not understood. This study aimed to investigate the role of NSUN7 in PCOS, particularly its impact on pyroptosis. KGN cells were treated with LPS to establish a cell model, and pyroptosis was evaluated using enzyme-linked immunosorbent assay, flow cytometry, and western blotting. The effect of NSUN7 on the m5C modification of NLRP3 was assessed using methylated RNA immunoprecipitation, dual-luciferase reporter assay, and actinomycin D treatment experiment. A PCOS rat model was generated, hormones were detected using ELISA, and pyroptosis was evaluated in the ovaries and primary granulosa cells. The results showed that NSUN7 expression was reduced in patients with PCOS and LPS-treated KGN cells. Silencing of NSUN7 inhibited LPS-induced cell pyroptosis. Moreover, NSUN7 knockdown inhibited m5C methylation of NLRP3 and reduced NLRP3 mRNA stability. Overexpression of NLRP3 reversed the pyroptosis inhibition induced by NSUN7 knockdown. Furthermore, interfering with NSUN7 alleviated hormone abnormalities and suppressed granulosa cell pyroptosis in PCOS rats. In conclusion, silencing of NSUN7 attenuates PCOS progression by inhibiting pyroptosis of granulosa cells by suppressing m5C methylation of NLRP3.
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高影响因子
iScience
·
2024
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IF 4.6
Hui Han,Yucong Sun,Wei Wei,Zixin Huang,Maosheng Cheng,Hongshen Qiu,Juan Wang,Siyi Zheng,Lianlian Liu,Qiang Zhang,Canfeng Zhang,Jieyi Ma,Siyao Guo,Zhaoyu Wang,Zhenpeng Li,Xu Jiang,Shuibin Lin,Qianwen Liu,Shuishen Zhang
Summary Emerging studies have demonstrated the link between RNA modifications and various cancers, while the predictive value and functional mechanisms of RNA modification-related genes (RMGs) in esophageal squamous cell carcinoma (ESCC) remain unclear. Here we established a prognostic signature for ESCC based on five RMGs. The analysis of ESCC clinical samples further verified the prognostic power of the prognostic signature. Moreover, we found that the knockdown of NSUN6 promotes ESCC progression in vitro and in vivo , whereas the overexpression of NSUN6 inhibits the malignant phenotype of ESCC cells. Mechanically, NSUN6 mediated tRNA m 5 C modifications selectively enhance the translation efficiency of CDH1 mRNA in a codon dependent manner. Rescue assays revealed that E-cadherin is an essential downstream target that mediates NSUN6’s function in the regulation of ESCC progression. These findings offer additional insights into the link between ESCC and RMGs, as well as provide potential strategies for ESCC management and therapy.
研究领域:
肿瘤学
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分子生物学
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基因组学
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DOI
PubMed
高影响因子
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