The methylation of DNA is a prevalent epigenetic modification that plays a crucial role in the pathological progression of ocular diseases.DNA methylation can regulate gene expression,thereby affecting cell function a...The methylation of DNA is a prevalent epigenetic modification that plays a crucial role in the pathological progression of ocular diseases.DNA methylation can regulate gene expression,thereby affecting cell function and signal transduction.Ophthalmic diseases are a kind of complex diseases,and their pathogenesis involves many factors such as genetic,environmental and individual differences.In addition,inflammation,oxidative stress and lipid metabolism,which abnormal DNA methylation is closely related to,are also considered to be major factors in eye diseases.The current understanding of DNA methylation in eye diseases is becoming more complex and comprehensive.In addition to the simple suppression of gene expression by hypermethylation,factors such as hypomethylation or demethylation,DNA methylation in non-promoter regions,interactions with other epigenetic modifications,and dynamic changes in DNA methylation must also be considered.Interestingly,although some genes are at abnormal methylation levels,their expression is not significantly changed,which indirectly reflects the complexity of gene regulation.This review aims to summarize and compare some relevant studies,and provide with new ideas and methods for the prevention and treatment of different eye diseases,such as glaucoma,retinoblastoma,and diabetic retinopathy.展开更多
As an essential transcriptional activator,PDX1 plays a crucial role in pancreatic development andβ-cell function.Mutations in the PDX1 gene may lead to type 4 maturityonset diabetes of the young(MODY4)and neonatal di...As an essential transcriptional activator,PDX1 plays a crucial role in pancreatic development andβ-cell function.Mutations in the PDX1 gene may lead to type 4 maturityonset diabetes of the young(MODY4)and neonatal diabetes mellitus.However,the precise mechanisms underlying MODY4 remain elusive due to the paucity of clinical samples and pronounced differences in pancreatic architecture and genomic composition between humans and existing animal models.In this study,three PDX1-mutant cynomolgus macaques were generated using CRISPR/Cas9 technology,all of which succumbed shortly postpartum,exhibiting pancreatic agenesis.Notably,one tri-allelic PDX1-mutant cynomolgus macaque(designated as M4)developed a pancreas,whereas the two monoallelic PDX1-mutant cynomolgus macaques displayed no anatomical evidence of pancreatic formation.RNA sequencing of the M4 pancreas revealed substantial molecular changes in both endocrine and exocrine functions,indicating developmental delay and PDX1haploinsufficiency.A marked change in m6A methylation was identified in the M4 pancreas,confirmed through cultured PDX1-mutantisletorganoids.Notably,overexpression of the m6A modulator METTL3 restored function in heterozygous PDX1-mutant islet organoids.This study highlights a novel role of m6A methylation modification in the progression of MODY4 and provides valuable molecular insights for preclinical research.展开更多
Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. ...Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. To explore the spatiotemporal expression of histone H3 lysine 4trimethylation（H3K4me3） and histone H3 lysine 27 trimethylation（H3K27me3） epigenetic marks and methylation or demethylation transferases in tooth organ development, we measured the expression of SET7, EZH2, KDM5 B and JMJD3 via immunohistochemistry and quantitative polymerase chain reaction（qP CR） analysis in the first molar of BALB/c mice embryos at E13.5, E15.5, E17.5, P0 and P3, respectively. We also measured the expression of H3K4me3 and H3K27me3 with immunofluorescence staining. During murine tooth germ development, methylation or demethylation transferases were expressed in a spatial–temporal manner. The bivalent modification characterized by H3K4me3 and H3K27me3 can be found during the tooth germ development, as shown by immunofluorescence. The expression of SET7, EZH2 as methylation transferases and KDM5 B and JMJD3 as demethylation transferases indicated accordingly with the expression of H3K4me3 and H3K27me3 respectively to some extent. The bivalent histone may play a critical role in tooth organ development via the regulation of cell differentiation.展开更多
Aims:Multiple genes and environmental factors are known to be involved in congenital heart disease(CHD),but epigenetic variation has received little attention.Monozygotic(MZ)twins with CHD provide a unique model for e...Aims:Multiple genes and environmental factors are known to be involved in congenital heart disease(CHD),but epigenetic variation has received little attention.Monozygotic(MZ)twins with CHD provide a unique model for exploring this phenomenon.In order to investigate the potential role of Deoxyribonucleic Acid(DNA)methyla-tion in CHD pathogenesis,the present study examined DNA methylation variation in MZ twins discordant for CHD,especially ventricular septal defect(VSD).Methods and Results:Using genome-wide DNA methylation profiles,we identified 4004 differentially methylated regions(DMRs)in 18 MZ twin pairs discordant for CHD,and 2826 genes were identified.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis revealed a list of CHD-associated pathways.To further investigate the role of DNA methylation in VSD,data from 7 pairs of MZ twins with VSD were analyzed.We identified 1614 DMRs corresponding to 1443 genes associated with arrhythmogenic right ventricular cardiomyopathy,cyclic guanosine monopho-sphate-protein kinase G(cGMP-PKG)signaling pathway by KEGG analysis,and cell-cell adhesion,calcium ion transmembrane transport by GO analysis.A proportion of DMR-associated genes were involved in calcium signaling pathways.The methylation changes of calcium signaling genes might be related to VSD pathogenesis.Conclusion:CHD is associated with differential DNA methylation in MZ twins.CHD may be etiologically linked to DNA methylation,and methylation of calcium signaling genes may be involved in the development of VSD.展开更多
microRNAs(miRNAs)are short single-stranded non-coding RNAs between 21 and 25 nt in length in eukaryotic organisms,which control post-transcriptional gene expression.Through complementary base pairing,miRNAs generally ...microRNAs(miRNAs)are short single-stranded non-coding RNAs between 21 and 25 nt in length in eukaryotic organisms,which control post-transcriptional gene expression.Through complementary base pairing,miRNAs generally bind to their target messenger RNAs and repress protein production by destabilizing the messenger RNA and translational silencing.They regulate almost all life activities,such as cell proliferation,differentiation,apoptosis,tumorigenesis,and host–pathogen interactions.Methylation modification is the most common RNA modification in eukaryotes.miRNA methylation exists in different types,mainly N6-methyladenosine,5-methylcytosine,and 7-methylguanine,which can change the expression level and biological mode of action of miRNAs and improve the activity of regulating gene expression in a very fine-tuned way with flexibility.In this review,we will summarize the recent findings concerning methylation modifications of miRNA,focusing on their biogenesis and the potential role of miRNA fate and functions.展开更多
Novel strategies for the simultaneous and portable detection of multiple analytes are highly favorable for clinical diagnosis and healthcare.Conventional colorimetric enzyme-linked immunosorbent assay(ELISA)is a widel...Novel strategies for the simultaneous and portable detection of multiple analytes are highly favorable for clinical diagnosis and healthcare.Conventional colorimetric enzyme-linked immunosorbent assay(ELISA)is a widely used laboratory technique for medical diagnostics,quality control,and research applications.However,nonspecific absorption of proteins may lead to a reduction of functional sites,resulting in high background and low sensitivity in ELISA.Herein,we report a simple method of functionalization of poly(methyl methacrylate)(PMMA)with polylysine to be used as the microfluidic microplate substrate for enhanced ELISA,enabling rapid,ultrasensitive,and multiplexed detection of infectious diseases.FTIR and fluorescence microscopy characterization confirmed high amine densities on polylysine-modified PMMA surface,resulting in high detection sensitivity of the colorimetric ELISA on the PMMA microdevice.The ultrasensitive polylysine-modified microplate can immobilize protein within 20 min and results of the assay can be viewed by the naked eye or scanned through a simple desktop scanner for quantitative analysis within 90 min.A sandwich-type immunoassay for the rapid and sensitive detection of immunoglobulin G(IgG),hepatitis B surface antigen(HBsAg),and hepatitis B core antigen(HBcAg)was demonstrated as a proof-of-concept for multiplexed detection.The limits of detection(LOD)of 200.0 pg/mL for IgG,180.0 pg/mL for HBsAg,and 300.0 pg/mL for HBcAg were achieved,without any specialized equipment like a microplate reader.The surface-modified microchip exhibited about 10-fold higher sensitivity than traditional microplates.This surface-modified microplate has tremendous potential as a point-ofcare multiplexed testing platform for many applications ranging from clinical diagnosis to environmental monitoring,particularly in resource-limited settings.展开更多
WntSa is a representative Wnt ligand that regulates multiple cellular functions through the Wnt5a nonclassical pathway.Although Wnt5a has been implicated in various pathological conditions,its role in cancer is ambigu...WntSa is a representative Wnt ligand that regulates multiple cellular functions through the Wnt5a nonclassical pathway.Although Wnt5a has been implicated in various pathological conditions,its role in cancer is ambiguous and might involve methyl modifications,distinct mRNA isofbrms,as well as different downstream pathways.Therefore,it is an essential factor in cancers'progression(invasion,migration,proliferation,and epithelial-mesenchymal transition),and a potential biomarker for prognosis and treatment.展开更多
Objective:Type 2 diabetes mellitus(T2DM)is characterized by insufficient insulin secretion and insulin resistance.Gypenosides(GPs)are the major bioactive saponins extracted from Gynostemma pentaphyllum.Previous studie...Objective:Type 2 diabetes mellitus(T2DM)is characterized by insufficient insulin secretion and insulin resistance.Gypenosides(GPs)are the major bioactive saponins extracted from Gynostemma pentaphyllum.Previous studies suggest that GPs have beneficial effects on T2DM,but the underlying mechanisms remain unclear.This study aims to investigate whether GPs exert therapeutic effects by influencing RNA N^(6)-methyladenosine(m6A)methylation modification,thereby regulating the downstream phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods:Expression levels and methylation changes of METTL3,METTL14,and FTO in T2DM were analyzed using public databases,and related pathways were explored via gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The main active components of GPs were screened from pharmacological databases,followed by compound-target network construction,enrichment analyses,and prediction of potential targets and pathways.A T2DM model was induced in Sprague-Dawley rats using a high-fat/high-sugar diet combined with low-dose streptozotocin.Rats were randomly divided into 4 groups:GPs-treated group(GPG,400 mg/kg),positive control group(PCG,metformin 100 mg/kg),normal saline control group(CONTROL),and T2DM model group(MODEL).Fasting blood glucose(FBG),oral glucose tolerance test(OGTT)-area under the glucose curve from 0 to 120 min(AUC0-120),fasting insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),serum inflammatory factors,and tissue pathology of pancreas and liver hematoxylin and eosin(HE)staining were assessed.Real-time fluorescence quantitative PCR and Western blotting were used to detect RNA and protein expression levels of METTL3,METTL14,FTO,PI3K,and AKT in pancreatic tissues.Molecular docking was applied to evaluate interactions between GPs’main components and METTL3,METTL14,and FTO to infer potential binding modes.Results:Bioinformatic analyses showed downregulation of METTL3/14 and upregulation of FTO in T2DM samples(all P<0.05),with enrichment in pathways related to insulin signaling,PI3K/AKT activation,oxidative stress response,and hormone secretion.Network pharmacology indicated that GPs components may act on targets involved in RNA modification and insulin-related pathways.In diabetic rats,GPs significantly reduced FBG,improved glucose tolerance,decreased HOMA-IR,and decreased the serum tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 levels compared with MODEL(all P<0.05).Pancreatic pathology showed alleviated islet injury and improved cell morphology.GPs treatment upregulated METTL3/14 mRNA and protein levels,and down-regulated FTO mRNA/protein levels in pancreatic tissue(all P<0.05).PI3K and AKT expression levels increased(both P<0.05),consistent with activation of downstream signals related to glucose uptake and improved insulin sensitivity.Metformin also improved metabolic indices but exhibited a different regulatory pattern on m6A-related enzymes compared with GPs.Molecular docking revealed stable interactions between core GPs saponin structures and methyltransferase-like 3(METTL3),methyltransferase-like 14(METTL14),or obesityassociated protein(FTO),suggesting that GPs may directly or indirectly modulate m6A regulatory proteins.Conclusion:GPs can effectively improve glucose-metabolism disorders,reduce inflammation,and protect pancreatic tissue in T2DM rats.The mechanisms may be associated with METTL3/14 up-regulation and FTO down-regulation,leading to enhanced m6A methylation and subsequent activation of the PI3K/AKT signaling pathway.These findings provide strong evidence for GPs regulation of epigenetic m6A RNA modification and insulin-related downstream pathways,and suggest that natural compounds targeting m6A regulation may be explored in the future for metabolic disease interventions.展开更多
In our previous studies, significant hypermethylation of the sirtuin 1（SIRT1） gene and demethylation of the b-amyloid precursor protein（APP） gene were found in patients with Alzheimer＇s disease（AD） compared wit...In our previous studies, significant hypermethylation of the sirtuin 1（SIRT1） gene and demethylation of the b-amyloid precursor protein（APP） gene were found in patients with Alzheimer＇s disease（AD） compared with the normal population. Moreover, the expression of SIRT1 was significantly decreased while that of APP was increased in AD patients. These results indicated a correlation of DNA methylation with gene expression levels in AD patients. To further investigate the epigenetic mechanism of gene modulation in AD, we used two epigenetic drugs, the DNA methylation inhibitor 5-aza-20-deoxycytidine（DAC） and the histone deacetylase inhibitor trichostatin A（TSA）, to treat human neuroblastoma SK-N-SH cells in the presence of amyloid b-peptide Ab25-35（Ab25-35）. We found that DAC and TSA had different effects on the expression trends of SIRT1 and APP in the cell model of amyloid toxicity. Although other genes, such as microtubule-associated protein s, presenilin 1, presenilin 2, and apolipoprotein E, were up-regulated after Ab25-35treatment, no significant differences were found after DAC and/or TSA treatment. These results support the evidence in AD patients and reveal a strong correlation of SIRT1/APP expression with DNA methylation and/or histone modification, which may help understand the pathogenesis of AD.展开更多
The global incidence of depression is progressively on the rise and tends to occur more in younger generations,however the pathogenesis of the disease is unclear.Meanwhile,epigenetics is a modification which produces ...The global incidence of depression is progressively on the rise and tends to occur more in younger generations,however the pathogenesis of the disease is unclear.Meanwhile,epigenetics is a modification which produces heritable alterations in the DNA sequence,which ultimately manifest in phenotypic differences.It has been suggested that the onset and development of depression can be tentatively explained by the combination of epigenetic and environmental factors.This paper reviews epigenetic changes in depression in the context of environmental factors,including DNA methylation modifications,histone modifications,and non-coding RNA regulation.An epigenetic-based therapeutic outlook was also proposed in this paper,which initially elucidates the epigenetic mechanisms underlying the pathogenesis of depressions and provides a theoretical basis for the treatment of depression.展开更多
Objective To explore the molecular mechanism by which curcumin affects renal interstitial fibrosis(RIF)progression by regulating ADAM metallopeptidase with thrombospondin type 1 motif 18(ADAMTS18)methylation.Methods N...Objective To explore the molecular mechanism by which curcumin affects renal interstitial fibrosis(RIF)progression by regulating ADAM metallopeptidase with thrombospondin type 1 motif 18(ADAMTS18)methylation.Methods NRK-49F cells RIF model were induced with transforming growth factorβ1(TGF-β1).Effects of different concentrations of curcumin(0,10,20,and 30μmol/L)on cell proliferation,cell cycle,cell apoptosis as well as cyclin D1 expression were analyzed by cell counting kit-8,flow cytometry and Western blot,respectively.ADAMTS18 methylation levels were determined by methylation-specific polymerase chain reaction.ADAMTS18,fibronectin(FN),type I collagen(Col-I)and alpha-smooth muscle actin(α-SMA)mRNA and protein expressions were analyzed by real-time PCR(RT-PCR)and Western blot,respectively.Meanwhile,cells were treated with 50 mmol/L 5-aza-2′-deoxycytidine(5-aza-dC,demethylation agent)for 72 h.Effect of curcumin on extracellular matrix(ECM)deposition was evaluated by immunochemical staining and Western blot.NRK-49F cells were transfected with ADAMTS18 small interfering RNA and grouped into a normal control,ADAMTS18-knock-out(KO),and ADAMTS18-KO+30μmol/L curcumin groups,and whether curcumin can reverse the effect of ADAMTS18 knockdown on RIF was evaluated.Results Compared with the control group,TGF-β1 significantly inhibited the proliferation of NRK-49F cells,blocked the G1/G0 phase,promoted cell apoptosis and inhibited cyclin D1 expression(P<0.01).Among the different concentrations of curcumin,30μmol/L curcumin significantly reversed these processes(P<0.01).Immunochemical staining and Western blot results showed that curcumin significantly inhibited the deposition of FN,Col-I andα-SMA(P<0.01).Curcumin and 5-zaz-dC had synergistic effects,promoting ADAMTS18 expression,removing ADAMTS18 methylation,and reducing ECM deposition.ADAMTS18 knockdown promoted ECM accumulation,and curcumin reversed this process(P<0.01).Conclusion TGF-β1-induced fibrosis in NRK-49F cells.Curcumin promoted ADAMTS18 expression,reduced ECM accumulation,and alleviated RIF progression by inhibiting ADAMTS18 methylation.展开更多
Post-transcriptional modifications,including histone modifications and DNA methylation,alter the chromatin landscape to regulate gene expression,thus control various cellular processes in plants.EARLY FLOWERING IN SHO...Post-transcriptional modifications,including histone modifications and DNA methylation,alter the chromatin landscape to regulate gene expression,thus control various cellular processes in plants.EARLY FLOWERING IN SHORT DAYS(EFS)is the major contributor for H3K36 methylation in Arabidopsis and is important for plant development.Here,we find that EFS is expressed in different stages of embryo morphogenesis,and the efs mutant produces larger embryo that results in enlarged seeds.Further analysis reveals that an imprinted gene MOP9.5 is hypomethylated at the promoter region and its expression is derepressed in efs mutant.MOP9.5 promoter is marked by various epigenetic modifications,and we find that following the increase of H3K36me3,H3K27me3 and H3K9me2 levels are reduced in efs mutant.This data indicates an antagonistic regulation between H3K36me3 and DNA methylation,and/or H3K27me3 at MOP9.5.Our results further show that both maternal and paternal EFS alleles are responsible for the seed size regulation,which unraveled a novel function of EFS in plant development.展开更多
文摘The methylation of DNA is a prevalent epigenetic modification that plays a crucial role in the pathological progression of ocular diseases.DNA methylation can regulate gene expression,thereby affecting cell function and signal transduction.Ophthalmic diseases are a kind of complex diseases,and their pathogenesis involves many factors such as genetic,environmental and individual differences.In addition,inflammation,oxidative stress and lipid metabolism,which abnormal DNA methylation is closely related to,are also considered to be major factors in eye diseases.The current understanding of DNA methylation in eye diseases is becoming more complex and comprehensive.In addition to the simple suppression of gene expression by hypermethylation,factors such as hypomethylation or demethylation,DNA methylation in non-promoter regions,interactions with other epigenetic modifications,and dynamic changes in DNA methylation must also be considered.Interestingly,although some genes are at abnormal methylation levels,their expression is not significantly changed,which indirectly reflects the complexity of gene regulation.This review aims to summarize and compare some relevant studies,and provide with new ideas and methods for the prevention and treatment of different eye diseases,such as glaucoma,retinoblastoma,and diabetic retinopathy.
基金supported by the National Key R&D Program of China(2018YFA0801404,2023YFC3403400)National Natural Science Foundation of China(81941006,32371190,32370878)+2 种基金Guangdong Special Support Program(2019BT02Y276,2024A1515012868)Double First-Class Discipline Promotion Project(2023B10564003)Program for Scientific Research Start-up Funds of Guangdong Ocean University(060302052408)。
文摘As an essential transcriptional activator,PDX1 plays a crucial role in pancreatic development andβ-cell function.Mutations in the PDX1 gene may lead to type 4 maturityonset diabetes of the young(MODY4)and neonatal diabetes mellitus.However,the precise mechanisms underlying MODY4 remain elusive due to the paucity of clinical samples and pronounced differences in pancreatic architecture and genomic composition between humans and existing animal models.In this study,three PDX1-mutant cynomolgus macaques were generated using CRISPR/Cas9 technology,all of which succumbed shortly postpartum,exhibiting pancreatic agenesis.Notably,one tri-allelic PDX1-mutant cynomolgus macaque(designated as M4)developed a pancreas,whereas the two monoallelic PDX1-mutant cynomolgus macaques displayed no anatomical evidence of pancreatic formation.RNA sequencing of the M4 pancreas revealed substantial molecular changes in both endocrine and exocrine functions,indicating developmental delay and PDX1haploinsufficiency.A marked change in m6A methylation was identified in the M4 pancreas,confirmed through cultured PDX1-mutantisletorganoids.Notably,overexpression of the m6A modulator METTL3 restored function in heterozygous PDX1-mutant islet organoids.This study highlights a novel role of m6A methylation modification in the progression of MODY4 and provides valuable molecular insights for preclinical research.
基金supported by National Science Foundation of China (Grant No. 81371136) to Xue-Dong ZhouNational Science Foundation of China (Grant No. 81200760 and 81470711) to Li-Wei Zheng
文摘Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. To explore the spatiotemporal expression of histone H3 lysine 4trimethylation（H3K4me3） and histone H3 lysine 27 trimethylation（H3K27me3） epigenetic marks and methylation or demethylation transferases in tooth organ development, we measured the expression of SET7, EZH2, KDM5 B and JMJD3 via immunohistochemistry and quantitative polymerase chain reaction（qP CR） analysis in the first molar of BALB/c mice embryos at E13.5, E15.5, E17.5, P0 and P3, respectively. We also measured the expression of H3K4me3 and H3K27me3 with immunofluorescence staining. During murine tooth germ development, methylation or demethylation transferases were expressed in a spatial–temporal manner. The bivalent modification characterized by H3K4me3 and H3K27me3 can be found during the tooth germ development, as shown by immunofluorescence. The expression of SET7, EZH2 as methylation transferases and KDM5 B and JMJD3 as demethylation transferases indicated accordingly with the expression of H3K4me3 and H3K27me3 respectively to some extent. The bivalent histone may play a critical role in tooth organ development via the regulation of cell differentiation.
基金China’s National Natural Science Foundation provided funding for this study(81900222)Guangzhou Science and Technology Program(SL2022A04J01269,202201020646)Guangzhou Health Science and Technology Program(20211A010026).
文摘Aims:Multiple genes and environmental factors are known to be involved in congenital heart disease(CHD),but epigenetic variation has received little attention.Monozygotic(MZ)twins with CHD provide a unique model for exploring this phenomenon.In order to investigate the potential role of Deoxyribonucleic Acid(DNA)methyla-tion in CHD pathogenesis,the present study examined DNA methylation variation in MZ twins discordant for CHD,especially ventricular septal defect(VSD).Methods and Results:Using genome-wide DNA methylation profiles,we identified 4004 differentially methylated regions(DMRs)in 18 MZ twin pairs discordant for CHD,and 2826 genes were identified.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis revealed a list of CHD-associated pathways.To further investigate the role of DNA methylation in VSD,data from 7 pairs of MZ twins with VSD were analyzed.We identified 1614 DMRs corresponding to 1443 genes associated with arrhythmogenic right ventricular cardiomyopathy,cyclic guanosine monopho-sphate-protein kinase G(cGMP-PKG)signaling pathway by KEGG analysis,and cell-cell adhesion,calcium ion transmembrane transport by GO analysis.A proportion of DMR-associated genes were involved in calcium signaling pathways.The methylation changes of calcium signaling genes might be related to VSD pathogenesis.Conclusion:CHD is associated with differential DNA methylation in MZ twins.CHD may be etiologically linked to DNA methylation,and methylation of calcium signaling genes may be involved in the development of VSD.
基金supported by the Natural Science Foundation of Jilin Province,China(No.20230101147JC)the National Natural Science Foundation of China(No.31970154)the China Postdoctoral Science Foundation(No.2022TQ0119).
文摘microRNAs(miRNAs)are short single-stranded non-coding RNAs between 21 and 25 nt in length in eukaryotic organisms,which control post-transcriptional gene expression.Through complementary base pairing,miRNAs generally bind to their target messenger RNAs and repress protein production by destabilizing the messenger RNA and translational silencing.They regulate almost all life activities,such as cell proliferation,differentiation,apoptosis,tumorigenesis,and host–pathogen interactions.Methylation modification is the most common RNA modification in eukaryotes.miRNA methylation exists in different types,mainly N6-methyladenosine,5-methylcytosine,and 7-methylguanine,which can change the expression level and biological mode of action of miRNAs and improve the activity of regulating gene expression in a very fine-tuned way with flexibility.In this review,we will summarize the recent findings concerning methylation modifications of miRNA,focusing on their biogenesis and the potential role of miRNA fate and functions.
基金support from NIH/NIAID(R41AI162477)the U.S.NSF(IIP2122712 and CHE2216473)+7 种基金the Cancer Prevention and Research Institute of Texas(CPRIT,RP210165)the TTUHSC-UTEP Joint Seed Grant,and the AAFS Foundation Research Lucas grantthe prior financial support to our research from the NIH/NIAID(R21AI107415)the NIH/NIGMS(SC2GM105584)the NIH/NIMHD RCMI Pilot grant(5G12MD007593-22)the NIH Building Scholar Summer Sabbatical Award,the NSF(IIP1953841,IIP2052347,and DMR1827745)the DOT(CARTEEH)the Philadelphia Foundation,the Medical Center of the Americas Foundation(MCA)。
文摘Novel strategies for the simultaneous and portable detection of multiple analytes are highly favorable for clinical diagnosis and healthcare.Conventional colorimetric enzyme-linked immunosorbent assay(ELISA)is a widely used laboratory technique for medical diagnostics,quality control,and research applications.However,nonspecific absorption of proteins may lead to a reduction of functional sites,resulting in high background and low sensitivity in ELISA.Herein,we report a simple method of functionalization of poly(methyl methacrylate)(PMMA)with polylysine to be used as the microfluidic microplate substrate for enhanced ELISA,enabling rapid,ultrasensitive,and multiplexed detection of infectious diseases.FTIR and fluorescence microscopy characterization confirmed high amine densities on polylysine-modified PMMA surface,resulting in high detection sensitivity of the colorimetric ELISA on the PMMA microdevice.The ultrasensitive polylysine-modified microplate can immobilize protein within 20 min and results of the assay can be viewed by the naked eye or scanned through a simple desktop scanner for quantitative analysis within 90 min.A sandwich-type immunoassay for the rapid and sensitive detection of immunoglobulin G(IgG),hepatitis B surface antigen(HBsAg),and hepatitis B core antigen(HBcAg)was demonstrated as a proof-of-concept for multiplexed detection.The limits of detection(LOD)of 200.0 pg/mL for IgG,180.0 pg/mL for HBsAg,and 300.0 pg/mL for HBcAg were achieved,without any specialized equipment like a microplate reader.The surface-modified microchip exhibited about 10-fold higher sensitivity than traditional microplates.This surface-modified microplate has tremendous potential as a point-ofcare multiplexed testing platform for many applications ranging from clinical diagnosis to environmental monitoring,particularly in resource-limited settings.
基金Fund supported by the Gansu Health and Family Planning Commission Funding(GSWSKY2018-34).
文摘WntSa is a representative Wnt ligand that regulates multiple cellular functions through the Wnt5a nonclassical pathway.Although Wnt5a has been implicated in various pathological conditions,its role in cancer is ambiguous and might involve methyl modifications,distinct mRNA isofbrms,as well as different downstream pathways.Therefore,it is an essential factor in cancers'progression(invasion,migration,proliferation,and epithelial-mesenchymal transition),and a potential biomarker for prognosis and treatment.
基金supported by College Students’Innovative Entrepreneurial Training Plan Program of Guizhou Province,China(S202210660105,S202310660055)。
文摘Objective:Type 2 diabetes mellitus(T2DM)is characterized by insufficient insulin secretion and insulin resistance.Gypenosides(GPs)are the major bioactive saponins extracted from Gynostemma pentaphyllum.Previous studies suggest that GPs have beneficial effects on T2DM,but the underlying mechanisms remain unclear.This study aims to investigate whether GPs exert therapeutic effects by influencing RNA N^(6)-methyladenosine(m6A)methylation modification,thereby regulating the downstream phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods:Expression levels and methylation changes of METTL3,METTL14,and FTO in T2DM were analyzed using public databases,and related pathways were explored via gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The main active components of GPs were screened from pharmacological databases,followed by compound-target network construction,enrichment analyses,and prediction of potential targets and pathways.A T2DM model was induced in Sprague-Dawley rats using a high-fat/high-sugar diet combined with low-dose streptozotocin.Rats were randomly divided into 4 groups:GPs-treated group(GPG,400 mg/kg),positive control group(PCG,metformin 100 mg/kg),normal saline control group(CONTROL),and T2DM model group(MODEL).Fasting blood glucose(FBG),oral glucose tolerance test(OGTT)-area under the glucose curve from 0 to 120 min(AUC0-120),fasting insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),serum inflammatory factors,and tissue pathology of pancreas and liver hematoxylin and eosin(HE)staining were assessed.Real-time fluorescence quantitative PCR and Western blotting were used to detect RNA and protein expression levels of METTL3,METTL14,FTO,PI3K,and AKT in pancreatic tissues.Molecular docking was applied to evaluate interactions between GPs’main components and METTL3,METTL14,and FTO to infer potential binding modes.Results:Bioinformatic analyses showed downregulation of METTL3/14 and upregulation of FTO in T2DM samples(all P<0.05),with enrichment in pathways related to insulin signaling,PI3K/AKT activation,oxidative stress response,and hormone secretion.Network pharmacology indicated that GPs components may act on targets involved in RNA modification and insulin-related pathways.In diabetic rats,GPs significantly reduced FBG,improved glucose tolerance,decreased HOMA-IR,and decreased the serum tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 levels compared with MODEL(all P<0.05).Pancreatic pathology showed alleviated islet injury and improved cell morphology.GPs treatment upregulated METTL3/14 mRNA and protein levels,and down-regulated FTO mRNA/protein levels in pancreatic tissue(all P<0.05).PI3K and AKT expression levels increased(both P<0.05),consistent with activation of downstream signals related to glucose uptake and improved insulin sensitivity.Metformin also improved metabolic indices but exhibited a different regulatory pattern on m6A-related enzymes compared with GPs.Molecular docking revealed stable interactions between core GPs saponin structures and methyltransferase-like 3(METTL3),methyltransferase-like 14(METTL14),or obesityassociated protein(FTO),suggesting that GPs may directly or indirectly modulate m6A regulatory proteins.Conclusion:GPs can effectively improve glucose-metabolism disorders,reduce inflammation,and protect pancreatic tissue in T2DM rats.The mechanisms may be associated with METTL3/14 up-regulation and FTO down-regulation,leading to enhanced m6A methylation and subsequent activation of the PI3K/AKT signaling pathway.These findings provide strong evidence for GPs regulation of epigenetic m6A RNA modification and insulin-related downstream pathways,and suggest that natural compounds targeting m6A regulation may be explored in the future for metabolic disease interventions.
基金supported by the National Basic Research Development Program of China (2006cb500700)the National Natural Science Foundation of China (30470904 and 31401627)+3 种基金Guangdong Provincial Natural Science Foundation of China (2010B031600070 and 2015A030313066)Science and Technology Social Development Project of Guangdong Province (2008B030301320 and 2012B031800053)a Guangdong Provincial Science and Technology Project (2013B051000009)a Guangzhou Science and Technology Plan Application Basic Research Project (2012J410076)
文摘In our previous studies, significant hypermethylation of the sirtuin 1（SIRT1） gene and demethylation of the b-amyloid precursor protein（APP） gene were found in patients with Alzheimer＇s disease（AD） compared with the normal population. Moreover, the expression of SIRT1 was significantly decreased while that of APP was increased in AD patients. These results indicated a correlation of DNA methylation with gene expression levels in AD patients. To further investigate the epigenetic mechanism of gene modulation in AD, we used two epigenetic drugs, the DNA methylation inhibitor 5-aza-20-deoxycytidine（DAC） and the histone deacetylase inhibitor trichostatin A（TSA）, to treat human neuroblastoma SK-N-SH cells in the presence of amyloid b-peptide Ab25-35（Ab25-35）. We found that DAC and TSA had different effects on the expression trends of SIRT1 and APP in the cell model of amyloid toxicity. Although other genes, such as microtubule-associated protein s, presenilin 1, presenilin 2, and apolipoprotein E, were up-regulated after Ab25-35treatment, no significant differences were found after DAC and/or TSA treatment. These results support the evidence in AD patients and reveal a strong correlation of SIRT1/APP expression with DNA methylation and/or histone modification, which may help understand the pathogenesis of AD.
文摘The global incidence of depression is progressively on the rise and tends to occur more in younger generations,however the pathogenesis of the disease is unclear.Meanwhile,epigenetics is a modification which produces heritable alterations in the DNA sequence,which ultimately manifest in phenotypic differences.It has been suggested that the onset and development of depression can be tentatively explained by the combination of epigenetic and environmental factors.This paper reviews epigenetic changes in depression in the context of environmental factors,including DNA methylation modifications,histone modifications,and non-coding RNA regulation.An epigenetic-based therapeutic outlook was also proposed in this paper,which initially elucidates the epigenetic mechanisms underlying the pathogenesis of depressions and provides a theoretical basis for the treatment of depression.
基金Supported by National Natural Science Foundation of China(No.82200740)。
文摘Objective To explore the molecular mechanism by which curcumin affects renal interstitial fibrosis(RIF)progression by regulating ADAM metallopeptidase with thrombospondin type 1 motif 18(ADAMTS18)methylation.Methods NRK-49F cells RIF model were induced with transforming growth factorβ1(TGF-β1).Effects of different concentrations of curcumin(0,10,20,and 30μmol/L)on cell proliferation,cell cycle,cell apoptosis as well as cyclin D1 expression were analyzed by cell counting kit-8,flow cytometry and Western blot,respectively.ADAMTS18 methylation levels were determined by methylation-specific polymerase chain reaction.ADAMTS18,fibronectin(FN),type I collagen(Col-I)and alpha-smooth muscle actin(α-SMA)mRNA and protein expressions were analyzed by real-time PCR(RT-PCR)and Western blot,respectively.Meanwhile,cells were treated with 50 mmol/L 5-aza-2′-deoxycytidine(5-aza-dC,demethylation agent)for 72 h.Effect of curcumin on extracellular matrix(ECM)deposition was evaluated by immunochemical staining and Western blot.NRK-49F cells were transfected with ADAMTS18 small interfering RNA and grouped into a normal control,ADAMTS18-knock-out(KO),and ADAMTS18-KO+30μmol/L curcumin groups,and whether curcumin can reverse the effect of ADAMTS18 knockdown on RIF was evaluated.Results Compared with the control group,TGF-β1 significantly inhibited the proliferation of NRK-49F cells,blocked the G1/G0 phase,promoted cell apoptosis and inhibited cyclin D1 expression(P<0.01).Among the different concentrations of curcumin,30μmol/L curcumin significantly reversed these processes(P<0.01).Immunochemical staining and Western blot results showed that curcumin significantly inhibited the deposition of FN,Col-I andα-SMA(P<0.01).Curcumin and 5-zaz-dC had synergistic effects,promoting ADAMTS18 expression,removing ADAMTS18 methylation,and reducing ECM deposition.ADAMTS18 knockdown promoted ECM accumulation,and curcumin reversed this process(P<0.01).Conclusion TGF-β1-induced fibrosis in NRK-49F cells.Curcumin promoted ADAMTS18 expression,reduced ECM accumulation,and alleviated RIF progression by inhibiting ADAMTS18 methylation.
基金supported by National Key R&D Program (2016YFA0500800)the National Natural Science Foundation of China (31571322)+2 种基金Tsinghua-Peking Joint Center for Life Sciences1000 Young Talent Program of ChinaS.Shafiq and Wei Xu are supported by the postdoctoral fellowships from Tsinghua-Peking Joint Center for Life Sciences
文摘Post-transcriptional modifications,including histone modifications and DNA methylation,alter the chromatin landscape to regulate gene expression,thus control various cellular processes in plants.EARLY FLOWERING IN SHORT DAYS(EFS)is the major contributor for H3K36 methylation in Arabidopsis and is important for plant development.Here,we find that EFS is expressed in different stages of embryo morphogenesis,and the efs mutant produces larger embryo that results in enlarged seeds.Further analysis reveals that an imprinted gene MOP9.5 is hypomethylated at the promoter region and its expression is derepressed in efs mutant.MOP9.5 promoter is marked by various epigenetic modifications,and we find that following the increase of H3K36me3,H3K27me3 and H3K9me2 levels are reduced in efs mutant.This data indicates an antagonistic regulation between H3K36me3 and DNA methylation,and/or H3K27me3 at MOP9.5.Our results further show that both maternal and paternal EFS alleles are responsible for the seed size regulation,which unraveled a novel function of EFS in plant development.
