How to effectively transform the pro-oncogenic tumor microenvironments(TME)surrounding a tumor into an anti-tumoral never fails to attract people to study.Small interfering RNA(siRNA)is considered one of the most note...How to effectively transform the pro-oncogenic tumor microenvironments(TME)surrounding a tumor into an anti-tumoral never fails to attract people to study.Small interfering RNA(siRNA)is considered one of the most noteworthy research directions that can regulate gene expression following a process known as RNA interference(RNAi).The research about siRNA delivery targeting tumor cells and TME has been on the rise in recent years.Using siRNA drugs to silence critical proteins in TME was one of the most efficient solutions.However,the manufacture of a siRNA delivery system faces three major obstacles,i.e.,appropriate cargo protection,accurately targeted delivery,and site-specific cargo release.In the following review,we summarized the pharmacological actions of siRNA drugs in remolding TME.In addition,the delivery strategies of siRNA drugs and combination therapy with siRNA drugs to remodel TME are thoroughly discussed.In the meanwhile,the most recent advancements in the development of all clinically investigated and commercialized siRNA delivery technologies are also presented.Ultimately,we propose that nanoparticle drug delivery siRNA may be the future research focus of oncogene therapy.This summary offers a thorough analysis and roadmap for general readers working in the field.展开更多
Cancer stem cells(CSCs) are a small subset of cells in cancers that are thought to initiate tumorous transformation and promote metastasis, recurrence, and resistance to treatment. Growing evidence has revealed the ex...Cancer stem cells(CSCs) are a small subset of cells in cancers that are thought to initiate tumorous transformation and promote metastasis, recurrence, and resistance to treatment. Growing evidence has revealed the existence of CSCs in various types of cancers and suggested that CSCs differentiate into diverse lineage cells that contribute to tumor progression. We may be able to overcome the limitations of cancer treatment with a comprehensive understanding of the biological features and mechanisms underlying therapeutic resistance in CSCs. This review provides an overview of the properties, biomarkers, and mechanisms of resistance shown by CSCs. Recent findings on metabolic features, especially fatty acid metabolism and ferroptosis in CSCs, are highlighted, along with promising targeting strategies. Targeting CSCs is a potential treatment plan to conquer cancer and prevent resistance and relapse in cancer treatment.展开更多
Glaucoma is a leading cause of irreve rsible blindness wo rldwide,and previous studies have shown that,in addition to affecting the eyes,it also causes abnormalities in the brain.However,it is not yet clear how the pr...Glaucoma is a leading cause of irreve rsible blindness wo rldwide,and previous studies have shown that,in addition to affecting the eyes,it also causes abnormalities in the brain.However,it is not yet clear how the primary visual cortex(V1)is altered in glaucoma.This study used DBA/2J mice as a model for spontaneous secondary glaucoma.The aim of the study was to compare the electrophysiological and histomorphological chara cteristics of neurons in the V1between 9-month-old DBA/2J mice and age-matched C57BL/6J mice.We conducted single-unit recordings in the V1 of light-anesthetized mice to measure the visually induced responses,including single-unit spiking and gamma band oscillations.The morphology of layerⅡ/Ⅲneurons was determined by neuronal nuclear antigen staining and Nissl staining of brain tissue sections.Eighty-seven neurons from eight DBA/2J mice and eighty-one neurons from eight C57BL/6J mice were examined.Compared with the C57BL/6J group,V1 neurons in the DBA/2J group exhibited weaker visual tuning and impaired spatial summation.Moreove r,fewer neuro ns were observed in the V1 of DBA/2J mice compared with C57BL/6J mice.These findings suggest that DBA/2J mice have fewer neurons in the VI compared with C57BL/6J mice,and that these neurons have impaired visual tuning.Our findings provide a better understanding of the pathological changes that occur in V1 neuron function and morphology in the DBA/2J mouse model.This study might offer some innovative perspectives regarding the treatment of glaucoma.展开更多
High myopia(HM)is the primary cause of blindness,with the microstructural organization and composition of collagenous fibers in the cornea and sclera playing a crucial role in the biomechanical behavior of these tissu...High myopia(HM)is the primary cause of blindness,with the microstructural organization and composition of collagenous fibers in the cornea and sclera playing a crucial role in the biomechanical behavior of these tissues.In a previously reported myopic linkage region,MYP5(17q21-22),a potential candidate gene,LRRC46(c.C235T,p.Q79X),was identified in a large Han Chinese pedigree.LRRC46 is expressed in various eye tissues in humans and mice,including the retina,cornea,and sclera.In subsequent cell experiments,the mutation(c.C235T)decreased the expression of LRRC46 protein in human corneal epithelial cells(HCE-T).Further investigation revealed that Lrrc46^(-/-)mice(KO)exhibited a classical myopia phenotype.The thickness of the cornea and sclera in KO mice became thinner and more pronounced with age,the activity of limbal stem cells decreased,and microstructural changes were observed in the fibroblasts of the sclera and cornea.We performed RNA-seq on scleral and corneal tissues of KO and normal control wild-type(WT)mice,which indicated a significant downregulation of the collagen synthesis-related pathway(extracellular matrix,ECM)in KO mice.Subsequent in vitro studies further indicated that LRRC46,a member of the important LRR protein family,primarily affected the formation of collagens.This study suggested that LRRC46 is a novel candidate gene for HM,influencing collagen protein VⅢ(Col8a1)formation in the eye and gradually altering the biomechanical structure of the cornea and sclera,thereby promoting the occurrence and development of HM.展开更多
Phosphatidylserine(PS)is distributed asymmetrically in the plasma membrane of eukaryotic cells.Phosphatidylserine flippase(P4-ATPase)transports PS from the outer leaflet of the lipid bilayer to the inner leaflet of th...Phosphatidylserine(PS)is distributed asymmetrically in the plasma membrane of eukaryotic cells.Phosphatidylserine flippase(P4-ATPase)transports PS from the outer leaflet of the lipid bilayer to the inner leaflet of the membrane to maintain PS asymmetry.TheβsubunitTMEM30 Ais indispensable for transport and proper function of P4-ATPase.Previous studies have shown that the ATP11 A and TMEM30 A complex is the molecular switch for myotube formation.However,the role of Tmem30 a in skeletal muscle regeneration remains elusive.In the current study,Tmem30 a was highly expressed in the tibialis anterior(TA)muscles of dystrophin-null(mdx)mice and BaCl2-induced muscle injury model mice.We generated a satellite cell(SC)-specific Tmem30 a conditional knockout(cKO)mouse model to investigate the role of Tmem30 a in skeletal muscle regeneration.The regenerative ability of cKO mice was evaluated by analyzing the number and diameter of regenerated SCs after the TA muscles were injured by BaCl2-injection.Compared to the control mice,the cKO mice showed decreased Pax7+and MYH3+SCs,indicating diminished SC proliferation,and decreased expression of muscular regulatory factors(MYOD and MYOG),suggesting impaired myoblast proliferation in skeletal muscle regeneration.Taken together,these results demonstrate the essential role of Tmem30 a in skeletal muscle regeneration.展开更多
Retinal angiogenesis is a critical process for normal retinal function.However,uncontrolled angiogenesis can lead to pathological neovascularization(NV),which is closely related to most irreversible blindness-causing ...Retinal angiogenesis is a critical process for normal retinal function.However,uncontrolled angiogenesis can lead to pathological neovascularization(NV),which is closely related to most irreversible blindness-causing retinal diseases.Understanding the molecular basis behind pathological NV is important for the treatment of related diseases.Twist-related protein 1(TWIST1)is a well-known transcription factor and principal inducer of epithelial-mesenchymal transition(EMT)in many human cancers.Our previous study showed that Twist1 expression is elevated in pathological retinal NV.To date,however,the role of TWIST1 in retinal pathological angiogenesis remains to be elucidated.To study the role of TWIST1 in pathological retinal NV and identify specific molecular targets for antagonizing pathological NV,we generated an inducible vascular endothelial cell(EC)-specific Twist1 transgenic mouse model(Tg-Twist1iEC+).Whole-mount retinas from Tg-Twist1iEC+mice showed retarded vascular progression and increased vascular density in the front end of the growing retinal vasculature,as well as aneurysm-like pathological retinal NV.Furthermore,overexpression of Twist1 in the ECs promoted cell proliferation but disturbed cell polarity,thus leading to uncontrolled retinal angiogenesis.TWIST1 promoted pathological NV by activating the Wnt/β-catenin signaling pathway and inducing the expression of NV formation-related genes,thereby acting as a‘valve’in the regulation of pathological angiogenesis.This study identified the critical role of TWIST1 in retinal pathological NV,thus providing a potential therapeutic target for pathological NV.展开更多
AIM：To investigate the effects of dexamethasone（DEX） and 1-（5-isoquinolinesulfonyl）-homopiperazine（HA1077） on actin cytoskeleton and β-catenin in cultured human trabecular meshwork（HTM） cells.METHODS： The H...AIM：To investigate the effects of dexamethasone（DEX） and 1-（5-isoquinolinesulfonyl）-homopiperazine（HA1077） on actin cytoskeleton and β-catenin in cultured human trabecular meshwork（HTM） cells.METHODS： The HTM cells were separated from human eyeball and cultured in vitro.They were divided into control group,DEX（1×10^-6mol/L） group,HA1077（3×10^-5mol/L）group,and DEX（1×10^-6mol/L） and HA1077（3×10^-5mol/L）group.Actin cytoskeleton and β-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses.RESULTS： In DEX group,there were reorganization of actin cytoskeleton and formation of cross linked actin networks（CLANs）,which were partially reversed in DEX and HA1077 group.DEX treatment also induced an increased expression of β-catenin,which was obviously reduced in DEX and HA1077 group.Meanwhile,the cultured HTM cells in HA1077 group had lower expression of β-catenin than that in the control group. CONCLUSION： Our results show that HA1077 can reverse the changes of actin organization and expression of β-catenin induced by DEX in cultured HTM cells,suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.展开更多
AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-compleme...AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.展开更多
Extensive effort has been devoted to the discovery,development,and validation of biomarkers for early disease diagnosis and prognosis as well as rapid evaluation of the response to therapeutic interventions.Genomic an...Extensive effort has been devoted to the discovery,development,and validation of biomarkers for early disease diagnosis and prognosis as well as rapid evaluation of the response to therapeutic interventions.Genomic and transcriptomic profiling are well-established means to identify disease-associated biomarkers.However,analysis of disease-associated peptidomes can also identify novel peptide biomarkers or signatures that provide sensitive and specific diagnostic and prognostic information for specific malignant,chronic,and infectious diseases.Growing evidence also suggests that peptidomic changes in liquid biopsies may more effectively detect changes in disease pathophysiology than other molecular methods.Knowledge gained from peptide-based diagnostic,therapeutic,and imaging approaches has led to promising new theranostic applications that can increase their bioavailability in target tissues at reduced doses to decrease side effects and improve treatment responses.However,despite major advances,multiple factors can still affect the utility of peptidomic data.This review summarizes several remaining challenges that affect peptide biomarker discovery and their use as diagnostics,with a focus on technological advances that can improve the detection,identification,and monitoring of peptide biomarkers for personalized medicine.展开更多
The endoplasmic reticulum(ER)membrane protein complex(EMC)is responsible for monitoring the biogenesis and synthetic quality of membrane proteins with tail-anchored or multiple transmembrane domains.The EMC subunit EM...The endoplasmic reticulum(ER)membrane protein complex(EMC)is responsible for monitoring the biogenesis and synthetic quality of membrane proteins with tail-anchored or multiple transmembrane domains.The EMC subunit EMC6 is one of the core members of EMC and forms an enclosed hydrophilic vestibule in cooperation with EMC3.Despite studies demonstrating that deletion of EMC3 led to rhodopsin mislocalization in rod photoreceptors of mice,the precise mechanism leading to the failure of rhodopsin trafficking remains unclear.Here,we generated the first rod photoreceptor-specific knockout of Emc6(RKO)and cone photoreceptor-specific knockout of Emc6(CKO)mouse models.Deficiency of Emc6 in rod photoreceptors led to progressive shortening of outer segments(OS),impaired visual function,mislocalization and reduced expression of rhodopsin,and increased gliosis in rod photoreceptors.In addition,CKO mice displayed the progressive death of cone photoreceptors and abnormal localization of cone opsin protein.Subsequently,proteomics analysis of the RKO mouse retina illustrated that several cilium-related proteins,particularly anoctamin-2(ANO2)and transmembrane protein 67(TMEM67),were significantly down-regulated prior to OS degeneration.Detrimental rod photoreceptor cilia and mislocalized membrane disc proteins were evident in RKO mice.Our data revealed that in addition to monitoring the synthesis of rhodopsin-dominated membrane disc proteins,EMC6 also impacted rod photoreceptors'ciliogenesis by regulating the synthesis of membrane proteins associated with cilia,contributing to the mislocalization of membrane disc proteins.展开更多
Dear Editor,Genome-wide association studies(GWASs)have reported a genetic association between certain populations and the severity of COVID-19.In severe patients,changes in the locus 3p21.31(including SLC6A20,LZTFL1,C...Dear Editor,Genome-wide association studies(GWASs)have reported a genetic association between certain populations and the severity of COVID-19.In severe patients,changes in the locus 3p21.31(including SLC6A20,LZTFL1,CCR9,FYCO1,CXCR6,and XCR1)are associated with COVID-19 hospitalization(Ellinghaus et al.,2020).Moreover,variations in 9p34.2 region(including genes related to the ABO blood grouping)can affect both susceptibility and severity of COVID-19;in particular,individuals with blood group A may have an increased risk of COVID-19 infection,while those with blood group O may have a protective effect(Ellinghaus et al.,2020;Shelton et al.,2021).展开更多
Blood vessel dysfunction causes several retinal diseases,including diabetic retinopathy,familial exudative vitreoretinopathy,macular degeneration and choroidal neovascularization in pathological myopia.Vascular endoth...Blood vessel dysfunction causes several retinal diseases,including diabetic retinopathy,familial exudative vitreoretinopathy,macular degeneration and choroidal neovascularization in pathological myopia.Vascular endothelial growth factor(VEGF)-neutralizing proteins provide benefits in most of those diseases,yet unsolved haemorrhage and frequent intraocular injections still bothered patients.Here,we identified endothelial CD146 as a new target for retinal diseases.CD146 expression was activated in two ocular pathological angiogenesis models,a laser-induced choroid neovascularization model and an oxygeninduced retinopathy model.The absence of CD146 impaired hypoxia-induced cell migration and angiogenesis both in cell lines and animal model.Preventive or therapeutic treatment with anti-CD146 antibody AA98 significantly inhibited hypoxia-induced aberrant retinal angiogenesis in two retinal disease models.Mechanistically,under hypoxia condition,CD146 was involved in the activation of NFκB,Erk and Akt signalling pathways,which are partially independent of VEGF.Consistently,anti-CD146therapy combined with anti-VEGF therapy showed enhanced impairment effect of hypoxia-induced angiogenesis in vitro and in vivo.Given the critical role of abnormal angiogenesis in retinal and choroidal diseases,our results provide novel insights into combinatorial therapy for neovascular fundus diseases.展开更多
Background:Fibroblast growth factor 10(FGF10)is implicated in the growth and development of the eye.Four singles nucleotide polymorphisms(SNPs)in the FGF10 gene(including rs1384449,rs339501,rs12517396 and rs10462070)w...Background:Fibroblast growth factor 10(FGF10)is implicated in the growth and development of the eye.Four singles nucleotide polymorphisms(SNPs)in the FGF10 gene(including rs1384449,rs339501,rs12517396 and rs10462070)were found to be associated with extreme myopia(EM,refractive error≤−10.0 diopters)in Japanese and Chinese Taiwan population.This case-control association study was conducted to explore the relationship between these four SNPs and high myopia in a western Chinese population.Methods:A total of 869 high myopia patients(HM,including 485 EM patients)and 899 healthy controls were recruited.These four SNPs were genotyped using the ABI SNaPshot method.Five genetic models(allelic,homozygous,heterozygous,dominant,and recessive)were applied to further evaluate the possible correlation between the SNPs and high myopia.The linkage-disequilibrium block(LD)structure was tested by Haploview Software.Results:In our study,no statistically significant differences were found between HM/EM patients and controls after Bonferroni multiple-correction(P>0.05)in the allele frequencies of these four SNPs in the FGF10 gene.We further found that rs12517396AA and rs10462070GG carriers showed a decreased risk of HM/EM compared with rs12517396AC+CC and rs10462070GA+AA carriers(P=0.045,OR=0.366;P=0.021,OR=0.131;P=0.03,OR=0.341;P=0.015,OR=0.122;respectively).Additionally,rs12517396AA and rs10462070GG carriers showed the same decreased risk of HM/EM compared with rs12517396CC and rs10462070AA carriers(P=0.048,OR=0.370;P=0.023,OR=0.133;P=0.032,OR=0.346;P=0.017,OR=0.126).However,these significant associations between rs12517396/rs10462070 and HM/EM disappeared after Bonferroni multiple-correction(P>0.05).Conclusion:Our findings indicate that rs12517396 and rs10462070 had marginal association with HM and EM.The other two common polymorphisms in FGF10 unlikely have significant effects in the genetic predisposition to HM/EM in western Chinese population.Further replication studies are needed to validate our findings in both animal models and human genetic epidemiologic studies.展开更多
Vogt–Koyanagi–Harada(VKH)disease is a leading cause of blindness in young and middle-aged people.However,the etiology of VKH disease remains unclear.Here,we performed the first trio-based whole-exome sequencing stud...Vogt–Koyanagi–Harada(VKH)disease is a leading cause of blindness in young and middle-aged people.However,the etiology of VKH disease remains unclear.Here,we performed the first trio-based whole-exome sequencing study,which enrolled 25 VKH patients and 50 controls,followed by a study of 2081 VKH patients from a Han Chinese population to uncover detrimental mutations.A total of 15 de novo mutations in VKH patients were identified,with one of the most important being the membrane palmitoylated protein 2(MPP2)p.K315N(MPP2-N315)mutation.The MPP2-N315 mutation was highly deleterious according to bioinformatic predictions.Additionally,this mutation appears rare,being absent from the 1000 Genome Project and Genome Aggregation Database,and it is highly conserved in 10 species,including humans and mice.Subsequent studies showed that pathological phenotypes and retinal vascular leakage were aggravated in MPP2-N315 mutation knock-in or MPP2-N315 adeno-associated virus-treated mice with experimental autoimmune uveitis(EAU).In vitro,we used clustered regularly interspaced short palindromic repeats(CRISPR‒Cas9)gene editing technology to delete intrinsic MPP2 before overexpressing wild-type MPP2 or MPP2-N315.Levels of cytokines,such as IL-1β,IL-17E,and vascular endothelial growth factor A,were increased,and barrier function was destroyed in the MPP2-N315 mutant ARPE19 cells.Mechanistically,the MPP2-N315 mutation had a stronger ability to directly bind to ANXA2 than MPP2-K315,as shown by LC‒MS/MS and Co-IP,and resulted in activation of the ERK3/IL-17E pathway.Overall,our results demonstrated that the MPP2-K315N mutation may increase susceptibility to VKH disease.展开更多
Mutations in serologically defined colon cancer autoantigen protein 8(SDCCAG8)were first identified in retinal ciliopathy families a decade ago with unknown function.To investigate the pathogenesis of SDCCAG8-associat...Mutations in serologically defined colon cancer autoantigen protein 8(SDCCAG8)were first identified in retinal ciliopathy families a decade ago with unknown function.To investigate the pathogenesis of SDCCAG8-associated retinal ciliopathies in vivo,we employed CRISPR/Cas9-mediated homology-directed recombination(HDR)to generate two knock-in mouse models,Sdccag8^(Y236X/Y236X) and Sdccag8^(E451GfsX467/E451GfsX467),which carry truncating mutations of the mouse Sdccag8,corresponding to mutations that cause Bardet-Biedl syndrome(BBS)and Senior-L?ken syndrome(SLS)(c.696T>G p.Y232X and c.1339-1340ins G p.E447GfsX463)in humans,respectively.The two mutant Sdccag8 knock-in mice faithfully recapitulated human SDCCAG8-associated BBS phenotypes such as rod-cone dystrophy,cystic renal disorder,polydactyly,infertility,and growth retardation,with varied age of onset and severity depending on the hypomorphic strength of the Sdccag8 mutations.To the best of our knowledge,these knock-in mouse lines are the first BBS mouse models to present with the polydactyly phenotype.Major phototransduction protein mislocalization was also observed outside the outer segment after initiation of photoreceptor degeneration.Impaired cilia were observed in the mutant photoreceptors,renal epithelial cells,and mouse embryonic fibroblasts derived from the knock-in mouse embryos,suggesting that SDCCAG8 plays an essential role in ciliogenesis,and cilium defects are a primary driving force of SDCCAG8-associated retinal ciliopathies.展开更多
The retinal pigment epithelium(RPE)and choroid are located behind the human retina and have multiple functions in the human visual system.Knowledge of the RPE and choroid cells and their gene expression profiles are f...The retinal pigment epithelium(RPE)and choroid are located behind the human retina and have multiple functions in the human visual system.Knowledge of the RPE and choroid cells and their gene expression profiles are fundamental for understanding retinal disease mechanisms and therapeutic strategies.Here,we sequenced the RNA of about 0.3 million single cells from human RPE and choroids across two regions and seven ages,revealing regional and age differences within the human RPE and choroid.Cell–cell interactions highlight the broad connectivity networks between the RPE and different choroid cell types.Moreover,the transcription factors and their target genes change during aging.The coding of somatic variations increases during aging in the human RPE and choroid at the single-cell level.Moreover,we identified ELN as a candidate for improving RPE degeneration and choroidal structure during aging.The mapping of the molecular architecture of the human RPE and choroid improves our understanding of the human vision support system and offers potential insights into the intervention targets for retinal diseases.展开更多
Endoplasmic reticulum(ER)membrane protein complex(EMC)is required for the co-translational insertion of newly synthesized multi-transmembrane proteins.Compromised EMC function in different cell types has been implicat...Endoplasmic reticulum(ER)membrane protein complex(EMC)is required for the co-translational insertion of newly synthesized multi-transmembrane proteins.Compromised EMC function in different cell types has been implicated in multiple diseases.Using inducible genetic mouse models,we revealed defects in retinal vascularization upon endothelial cell(EC)specific deletion of Emc1,the largest subunit of EMC.Loss of Emc1 in ECs led to reduced vascular progression and vascular density,diminished tip cell sprouts,and vascular leakage.We then performed an unbiased transcriptomic analysis on human retinal microvascular endothelial cells(HRECs)and revealed a pivotal role of EMC1 in theβ-catenin signaling pathway.Further in-vitro and in-vivo experiments proved that loss of EMC1 led to compromisedβ-catenin signaling activity through reduced expression of Wnt receptor FZD4,which could be restored by lithium chloride(LiCl)treatment.Driven by these findings,we screened genomic DNA samples from familial exudative vitreoretinopathy(FEVR)patients and identified one heterozygous variant in EMC1 that co-segregated with FEVR phenotype in the family.In-vitro expression experiments revealed that this variant allele failed to facilitate the expression of FZD4 on the plasma membrane and activate theβ-catenin signaling pathway,which might be a main cause of FEVR.In conclusion,our findings reveal that variants in EMC1 gene cause compromisedβ-catenin signaling activity,which may be associated with the pathogenesis of FEVR.展开更多
AIM:To systematically understand the genetic association between methylenetetrahydrofolate reductase(MTHFR)C677 T polymorphism and primary open angle glaucoma(POAG).METHODS:A comprehensive literature search in Google ...AIM:To systematically understand the genetic association between methylenetetrahydrofolate reductase(MTHFR)C677 T polymorphism and primary open angle glaucoma(POAG).METHODS:A comprehensive literature search in Google Scholar,Pub Med,Science Citation Index,Foreign Medical Literature Retrieval Service,Chinese National Knowledge Infrastructure and Wanfang Databases was performed to collect all eligible studies up to August 2019.Study selection,data abstraction and study quality evaluation were performed by two independent investigators.Odds ratios(ORs)with 95%confidence intervals(CIs)were calculated to assess the association.RESULTS:Eighteen case-control studies including 2156 cases and 2201 controls were identified.There was no significant difference in the terms of MTHFR C677 T polymorphism and POAG in the Caucasian population(for T vs C OR=1.11,95%CI:0.88 to 1.39;for TT vs CC OR=1.01,95%CI:0.76 to 1.36;for TT+TC vs CC OR=1.15,95%CI:0.84 to 1.58 and for TT vs TC+CC OR=1.02,95%CI:0.78 to 1.33).However,a significant effect was revealed in the Asian population(for T vs C OR=1.34,95%CI:1.12 to 1.59;for TT+TC vs CC OR=1.41,95%CI:1.14 to 1.76).CONCLUSION:Based on 18 eligible studies,we provide a correlation between MTHFR C677 T polymorphism and POAG among the Asians subgroup indicating that the T allele or TT+TC genotype may play a critical role in POAG development in Asians.展开更多
Critical coronavirus disease 2019(COVID-19)is associated with high mortality and potential genetic factors have been reported to be involved in the development of critical COVID-19.We performed a genome-wide associati...Critical coronavirus disease 2019(COVID-19)is associated with high mortality and potential genetic factors have been reported to be involved in the development of critical COVID-19.We performed a genome-wide association study to identify the genetic factors responsible for developing critical COVID-19.632 critical patients with COVID-19 and 3021 healthy controls from the Chinese population were recruited.First,we identified a genome-wide significant difference of IL-6 rs2069837(p=9.73×10^(−15),OR=0.41)between 437 critical patients with COVID-19 and 2551 normal controls in the discovery cohort.展开更多
Inherited retinal dystrophies (IRDs) are major causes of visual impairment and irreversible blindness worldwide, while the precise molecular and genetic mechanisms are still elusive. N6-methyladenosine (m^(6)A) modifi...Inherited retinal dystrophies (IRDs) are major causes of visual impairment and irreversible blindness worldwide, while the precise molecular and genetic mechanisms are still elusive. N6-methyladenosine (m^(6)A) modification is the most prevalent internal modification in eukaryotic mRNA. YTH domain containing 2 (YTHDC2), an m^(6)A reader protein, has recently been identified as a key player in germline development and human cancer. However, its contribution to retinal function remains unknown. Here, we explore the role of YTHDC2 in the visual function of retinal rod photoreceptors by generating rod-specific Ythdc2 knockout mice. Results show that Ythdc2 deficiency in rods causes diminished scotopic ERG responses and progressive retinal degeneration. Multi-omics analysis further identifies Ppef2 and Pde6b as the potential targets of YTHDC2 in the retina. Specifically, via its YTH domain, YTHDC2 recognizes and binds m^(6)A-modified Ppef2 mRNA at the coding sequence and Pde6b mRNA at the 5′-UTR, resulting in enhanced translation efficiency without affecting mRNA levels. Compromised translation efficiency of Ppef2 and Pde6b after YTHDC2 depletion ultimately leads to decreased protein levels in the retina, impaired retinal function, and progressive rod death. Collectively, our finding highlights the importance of YTHDC2 in visual function and photoreceptor survival, which provides an unreported elucidation of IRD pathogenesis via epitranscriptomics.展开更多
基金supported by the Natural Science Foundation of China(U2230123,81870683,82121003 and 82201234)CAMS Innovation Fund for Medical Sciences(2019-I2M-5-032)the Department of Science and Technology of Sichuan Province,China(22ZYZYTS0159,2022YFS0606,2023YFS0125,2023YFS0131,2023NSFSC0033,and 22ZYZYTS0151)
文摘How to effectively transform the pro-oncogenic tumor microenvironments(TME)surrounding a tumor into an anti-tumoral never fails to attract people to study.Small interfering RNA(siRNA)is considered one of the most noteworthy research directions that can regulate gene expression following a process known as RNA interference(RNAi).The research about siRNA delivery targeting tumor cells and TME has been on the rise in recent years.Using siRNA drugs to silence critical proteins in TME was one of the most efficient solutions.However,the manufacture of a siRNA delivery system faces three major obstacles,i.e.,appropriate cargo protection,accurately targeted delivery,and site-specific cargo release.In the following review,we summarized the pharmacological actions of siRNA drugs in remolding TME.In addition,the delivery strategies of siRNA drugs and combination therapy with siRNA drugs to remodel TME are thoroughly discussed.In the meanwhile,the most recent advancements in the development of all clinically investigated and commercialized siRNA delivery technologies are also presented.Ultimately,we propose that nanoparticle drug delivery siRNA may be the future research focus of oncogene therapy.This summary offers a thorough analysis and roadmap for general readers working in the field.
基金supported by the National Key Research and Development Program of China (Grant No.2023YFC3402100)the National Natural Science Foundation of China (Grant No.92259102)。
文摘Cancer stem cells(CSCs) are a small subset of cells in cancers that are thought to initiate tumorous transformation and promote metastasis, recurrence, and resistance to treatment. Growing evidence has revealed the existence of CSCs in various types of cancers and suggested that CSCs differentiate into diverse lineage cells that contribute to tumor progression. We may be able to overcome the limitations of cancer treatment with a comprehensive understanding of the biological features and mechanisms underlying therapeutic resistance in CSCs. This review provides an overview of the properties, biomarkers, and mechanisms of resistance shown by CSCs. Recent findings on metabolic features, especially fatty acid metabolism and ferroptosis in CSCs, are highlighted, along with promising targeting strategies. Targeting CSCs is a potential treatment plan to conquer cancer and prevent resistance and relapse in cancer treatment.
基金supported by the STI 2030-Major Projects 2022ZD0208500(to DY)the National Natural Science Foundation of China,Nos.82072011(to YX),82121003(to DY),82271120(to YS)+2 种基金Sichuan Science and Technology Program,No.2022ZYD0066(to YS)a grant from Chinese Academy of Medical Science,No.2019-12M-5-032(to YS)the Fundamental Research Funds for the Central Universities,No.ZYGX2021YGLH219(to KC)。
文摘Glaucoma is a leading cause of irreve rsible blindness wo rldwide,and previous studies have shown that,in addition to affecting the eyes,it also causes abnormalities in the brain.However,it is not yet clear how the primary visual cortex(V1)is altered in glaucoma.This study used DBA/2J mice as a model for spontaneous secondary glaucoma.The aim of the study was to compare the electrophysiological and histomorphological chara cteristics of neurons in the V1between 9-month-old DBA/2J mice and age-matched C57BL/6J mice.We conducted single-unit recordings in the V1 of light-anesthetized mice to measure the visually induced responses,including single-unit spiking and gamma band oscillations.The morphology of layerⅡ/Ⅲneurons was determined by neuronal nuclear antigen staining and Nissl staining of brain tissue sections.Eighty-seven neurons from eight DBA/2J mice and eighty-one neurons from eight C57BL/6J mice were examined.Compared with the C57BL/6J group,V1 neurons in the DBA/2J group exhibited weaker visual tuning and impaired spatial summation.Moreove r,fewer neuro ns were observed in the V1 of DBA/2J mice compared with C57BL/6J mice.These findings suggest that DBA/2J mice have fewer neurons in the VI compared with C57BL/6J mice,and that these neurons have impaired visual tuning.Our findings provide a better understanding of the pathological changes that occur in V1 neuron function and morphology in the DBA/2J mouse model.This study might offer some innovative perspectives regarding the treatment of glaucoma.
基金supported by the National Natural Science Foundation of China(82330030,82271120,82121003,82201234)the CAMS Innovation Fund for Medical Sciences(2019-12 M-5-032,2021LY06)Sichuan Science and Technology Program(2021YFS0369,2022ZYD0131,24YSZH0012,23ZYZYTS0271,TB2023093,2023ZY0059)。
文摘High myopia(HM)is the primary cause of blindness,with the microstructural organization and composition of collagenous fibers in the cornea and sclera playing a crucial role in the biomechanical behavior of these tissues.In a previously reported myopic linkage region,MYP5(17q21-22),a potential candidate gene,LRRC46(c.C235T,p.Q79X),was identified in a large Han Chinese pedigree.LRRC46 is expressed in various eye tissues in humans and mice,including the retina,cornea,and sclera.In subsequent cell experiments,the mutation(c.C235T)decreased the expression of LRRC46 protein in human corneal epithelial cells(HCE-T).Further investigation revealed that Lrrc46^(-/-)mice(KO)exhibited a classical myopia phenotype.The thickness of the cornea and sclera in KO mice became thinner and more pronounced with age,the activity of limbal stem cells decreased,and microstructural changes were observed in the fibroblasts of the sclera and cornea.We performed RNA-seq on scleral and corneal tissues of KO and normal control wild-type(WT)mice,which indicated a significant downregulation of the collagen synthesis-related pathway(extracellular matrix,ECM)in KO mice.Subsequent in vitro studies further indicated that LRRC46,a member of the important LRR protein family,primarily affected the formation of collagens.This study suggested that LRRC46 is a novel candidate gene for HM,influencing collagen protein VⅢ(Col8a1)formation in the eye and gradually altering the biomechanical structure of the cornea and sclera,thereby promoting the occurrence and development of HM.
基金supported by the National Natural Science Foundation of China(81770950,81970841)Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(2019-12M-5-032)Department of Science and Technology of Sichuan Province(21ZDYF4279,2020JDZH0026,2021JDZH0022)。
文摘Phosphatidylserine(PS)is distributed asymmetrically in the plasma membrane of eukaryotic cells.Phosphatidylserine flippase(P4-ATPase)transports PS from the outer leaflet of the lipid bilayer to the inner leaflet of the membrane to maintain PS asymmetry.TheβsubunitTMEM30 Ais indispensable for transport and proper function of P4-ATPase.Previous studies have shown that the ATP11 A and TMEM30 A complex is the molecular switch for myotube formation.However,the role of Tmem30 a in skeletal muscle regeneration remains elusive.In the current study,Tmem30 a was highly expressed in the tibialis anterior(TA)muscles of dystrophin-null(mdx)mice and BaCl2-induced muscle injury model mice.We generated a satellite cell(SC)-specific Tmem30 a conditional knockout(cKO)mouse model to investigate the role of Tmem30 a in skeletal muscle regeneration.The regenerative ability of cKO mice was evaluated by analyzing the number and diameter of regenerated SCs after the TA muscles were injured by BaCl2-injection.Compared to the control mice,the cKO mice showed decreased Pax7+and MYH3+SCs,indicating diminished SC proliferation,and decreased expression of muscular regulatory factors(MYOD and MYOG),suggesting impaired myoblast proliferation in skeletal muscle regeneration.Taken together,these results demonstrate the essential role of Tmem30 a in skeletal muscle regeneration.
基金supported by the National Natural Science Foundation of China(82071009,81700841)the Grant from Chinese Academy of Medical Sciences(2019-I2M-5-032)。
文摘Retinal angiogenesis is a critical process for normal retinal function.However,uncontrolled angiogenesis can lead to pathological neovascularization(NV),which is closely related to most irreversible blindness-causing retinal diseases.Understanding the molecular basis behind pathological NV is important for the treatment of related diseases.Twist-related protein 1(TWIST1)is a well-known transcription factor and principal inducer of epithelial-mesenchymal transition(EMT)in many human cancers.Our previous study showed that Twist1 expression is elevated in pathological retinal NV.To date,however,the role of TWIST1 in retinal pathological angiogenesis remains to be elucidated.To study the role of TWIST1 in pathological retinal NV and identify specific molecular targets for antagonizing pathological NV,we generated an inducible vascular endothelial cell(EC)-specific Twist1 transgenic mouse model(Tg-Twist1iEC+).Whole-mount retinas from Tg-Twist1iEC+mice showed retarded vascular progression and increased vascular density in the front end of the growing retinal vasculature,as well as aneurysm-like pathological retinal NV.Furthermore,overexpression of Twist1 in the ECs promoted cell proliferation but disturbed cell polarity,thus leading to uncontrolled retinal angiogenesis.TWIST1 promoted pathological NV by activating the Wnt/β-catenin signaling pathway and inducing the expression of NV formation-related genes,thereby acting as a‘valve’in the regulation of pathological angiogenesis.This study identified the critical role of TWIST1 in retinal pathological NV,thus providing a potential therapeutic target for pathological NV.
基金Supported by the Natural Science Foundation of China (No.81300768 No.81371048+4 种基金 No.81670853)Science and Technology Department of Sichuan Province of China (No.2015HH0031)Health and Family Planning Commission of Sichuan Province of China (No.100539 No. 090505 No.090514)
文摘AIM：To investigate the effects of dexamethasone（DEX） and 1-（5-isoquinolinesulfonyl）-homopiperazine（HA1077） on actin cytoskeleton and β-catenin in cultured human trabecular meshwork（HTM） cells.METHODS： The HTM cells were separated from human eyeball and cultured in vitro.They were divided into control group,DEX（1×10^-6mol/L） group,HA1077（3×10^-5mol/L）group,and DEX（1×10^-6mol/L） and HA1077（3×10^-5mol/L）group.Actin cytoskeleton and β-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses.RESULTS： In DEX group,there were reorganization of actin cytoskeleton and formation of cross linked actin networks（CLANs）,which were partially reversed in DEX and HA1077 group.DEX treatment also induced an increased expression of β-catenin,which was obviously reduced in DEX and HA1077 group.Meanwhile,the cultured HTM cells in HA1077 group had lower expression of β-catenin than that in the control group. CONCLUSION： Our results show that HA1077 can reverse the changes of actin organization and expression of β-catenin induced by DEX in cultured HTM cells,suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.
基金Supported by the National Natural Science Foundation of China(No.81900829,No.82070963)the Xiamen Medical and Health Guiding Project Fund Project(No.3502Z20214ZD1214)+1 种基金the Guangdong Basic and Applied Basic Research Foundation(No.2019A1515011234)the Science and Technology Innovation Committee of Shenzhen(No.JCYJ20210324125614039)。
文摘AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.
基金The work was primarily supported by research funding provided by the National Cancer Institute(U01CA252965)Eunice Kennedy Shriver National Institute of Child Health and Human Development(R01HD090927 and R01HD103511)+2 种基金National Institute of Allergy and Infectious Diseases(R01AI144168)U.S.Department of Defense(W8IXWH1910926)National Institute of Neurological Disorders and Stroke(R21NS130542).
文摘Extensive effort has been devoted to the discovery,development,and validation of biomarkers for early disease diagnosis and prognosis as well as rapid evaluation of the response to therapeutic interventions.Genomic and transcriptomic profiling are well-established means to identify disease-associated biomarkers.However,analysis of disease-associated peptidomes can also identify novel peptide biomarkers or signatures that provide sensitive and specific diagnostic and prognostic information for specific malignant,chronic,and infectious diseases.Growing evidence also suggests that peptidomic changes in liquid biopsies may more effectively detect changes in disease pathophysiology than other molecular methods.Knowledge gained from peptide-based diagnostic,therapeutic,and imaging approaches has led to promising new theranostic applications that can increase their bioavailability in target tissues at reduced doses to decrease side effects and improve treatment responses.However,despite major advances,multiple factors can still affect the utility of peptidomic data.This review summarizes several remaining challenges that affect peptide biomarker discovery and their use as diagnostics,with a focus on technological advances that can improve the detection,identification,and monitoring of peptide biomarkers for personalized medicine.
基金supported by The National Natural Science Foundation of China(No.82121003,81970841,82101160)the program of Science and Technology International Cooperation Project of Qinghai province(China)(No.2022-HZ-814)+2 种基金the CAMS Innovation Fund for Medical Sciences(No.2019-12M-5-032)Sichuan Intellectual Property Office(China)(No.2022-ZS-0070)the Department of Chengdu Science and Technology(Sichuan,China)(No.2021-YF05-01316-SN).
文摘The endoplasmic reticulum(ER)membrane protein complex(EMC)is responsible for monitoring the biogenesis and synthetic quality of membrane proteins with tail-anchored or multiple transmembrane domains.The EMC subunit EMC6 is one of the core members of EMC and forms an enclosed hydrophilic vestibule in cooperation with EMC3.Despite studies demonstrating that deletion of EMC3 led to rhodopsin mislocalization in rod photoreceptors of mice,the precise mechanism leading to the failure of rhodopsin trafficking remains unclear.Here,we generated the first rod photoreceptor-specific knockout of Emc6(RKO)and cone photoreceptor-specific knockout of Emc6(CKO)mouse models.Deficiency of Emc6 in rod photoreceptors led to progressive shortening of outer segments(OS),impaired visual function,mislocalization and reduced expression of rhodopsin,and increased gliosis in rod photoreceptors.In addition,CKO mice displayed the progressive death of cone photoreceptors and abnormal localization of cone opsin protein.Subsequently,proteomics analysis of the RKO mouse retina illustrated that several cilium-related proteins,particularly anoctamin-2(ANO2)and transmembrane protein 67(TMEM67),were significantly down-regulated prior to OS degeneration.Detrimental rod photoreceptor cilia and mislocalized membrane disc proteins were evident in RKO mice.Our data revealed that in addition to monitoring the synthesis of rhodopsin-dominated membrane disc proteins,EMC6 also impacted rod photoreceptors'ciliogenesis by regulating the synthesis of membrane proteins associated with cilia,contributing to the mislocalization of membrane disc proteins.
基金supported by the National Natural Science Foundation of China(81790643,82121003,and 82271105)the Sichuan Science and Technology Program(2019YFS0368,2021YFS0404,2021YFS0369,2021YFS0388,2021ZYD0082,2021YFS0033,and 2022NSFSC1352)+2 种基金the CAMS Innovation Fund for Medical Sciences(2019-12M-5-032)Sichuan Provincial People's Hospital(2021QN13)2021 open project of Sichuan Provincial Key Laboratory for Human Disease Genetics Research(2021kflx008).
文摘Dear Editor,Genome-wide association studies(GWASs)have reported a genetic association between certain populations and the severity of COVID-19.In severe patients,changes in the locus 3p21.31(including SLC6A20,LZTFL1,CCR9,FYCO1,CXCR6,and XCR1)are associated with COVID-19 hospitalization(Ellinghaus et al.,2020).Moreover,variations in 9p34.2 region(including genes related to the ABO blood grouping)can affect both susceptibility and severity of COVID-19;in particular,individuals with blood group A may have an increased risk of COVID-19 infection,while those with blood group O may have a protective effect(Ellinghaus et al.,2020;Shelton et al.,2021).
基金supported by the CAMS Innovation Fund for Medical Sciences (2019-12M-5-032)the National Natural Science Foundation of China (81790643,82121003 to ZY,and 31770793)the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2018122)。
文摘Blood vessel dysfunction causes several retinal diseases,including diabetic retinopathy,familial exudative vitreoretinopathy,macular degeneration and choroidal neovascularization in pathological myopia.Vascular endothelial growth factor(VEGF)-neutralizing proteins provide benefits in most of those diseases,yet unsolved haemorrhage and frequent intraocular injections still bothered patients.Here,we identified endothelial CD146 as a new target for retinal diseases.CD146 expression was activated in two ocular pathological angiogenesis models,a laser-induced choroid neovascularization model and an oxygeninduced retinopathy model.The absence of CD146 impaired hypoxia-induced cell migration and angiogenesis both in cell lines and animal model.Preventive or therapeutic treatment with anti-CD146 antibody AA98 significantly inhibited hypoxia-induced aberrant retinal angiogenesis in two retinal disease models.Mechanistically,under hypoxia condition,CD146 was involved in the activation of NFκB,Erk and Akt signalling pathways,which are partially independent of VEGF.Consistently,anti-CD146therapy combined with anti-VEGF therapy showed enhanced impairment effect of hypoxia-induced angiogenesis in vitro and in vivo.Given the critical role of abnormal angiogenesis in retinal and choroidal diseases,our results provide novel insights into combinatorial therapy for neovascular fundus diseases.
基金supported by the Top-Notch Young Talents Program of China(to Y.S.)Natural Science Foundation of China(81570888 and 81870683 to Y.S.+2 种基金81400401 to Y.Y.)National Key Scientific Research Program(2017YFC0907302 to Y.S.)Department of Science and Technology of Sichuan Province,China(2015SZ0242 and 2016JQ0026 to Y.S.).
文摘Background:Fibroblast growth factor 10(FGF10)is implicated in the growth and development of the eye.Four singles nucleotide polymorphisms(SNPs)in the FGF10 gene(including rs1384449,rs339501,rs12517396 and rs10462070)were found to be associated with extreme myopia(EM,refractive error≤−10.0 diopters)in Japanese and Chinese Taiwan population.This case-control association study was conducted to explore the relationship between these four SNPs and high myopia in a western Chinese population.Methods:A total of 869 high myopia patients(HM,including 485 EM patients)and 899 healthy controls were recruited.These four SNPs were genotyped using the ABI SNaPshot method.Five genetic models(allelic,homozygous,heterozygous,dominant,and recessive)were applied to further evaluate the possible correlation between the SNPs and high myopia.The linkage-disequilibrium block(LD)structure was tested by Haploview Software.Results:In our study,no statistically significant differences were found between HM/EM patients and controls after Bonferroni multiple-correction(P>0.05)in the allele frequencies of these four SNPs in the FGF10 gene.We further found that rs12517396AA and rs10462070GG carriers showed a decreased risk of HM/EM compared with rs12517396AC+CC and rs10462070GA+AA carriers(P=0.045,OR=0.366;P=0.021,OR=0.131;P=0.03,OR=0.341;P=0.015,OR=0.122;respectively).Additionally,rs12517396AA and rs10462070GG carriers showed the same decreased risk of HM/EM compared with rs12517396CC and rs10462070AA carriers(P=0.048,OR=0.370;P=0.023,OR=0.133;P=0.032,OR=0.346;P=0.017,OR=0.126).However,these significant associations between rs12517396/rs10462070 and HM/EM disappeared after Bonferroni multiple-correction(P>0.05).Conclusion:Our findings indicate that rs12517396 and rs10462070 had marginal association with HM and EM.The other two common polymorphisms in FGF10 unlikely have significant effects in the genetic predisposition to HM/EM in western Chinese population.Further replication studies are needed to validate our findings in both animal models and human genetic epidemiologic studies.
基金We thank the families for participation in this study,and we thank Novogene Technology Co.,Ltd.,for the WES sequencing and analysis.This work was supported by the National Natural Science Foundation Project of China(82070951,82271078)the National Natural Science Foundation Key Program(81930023)+3 种基金The Innovative Research Group Project of Chongqing Education Commission(CXQT19015)the Innovation Supporting Plan of Overseas Study of Chongqing(cx2018010)the National Key Clinical Specialties Construction Program of China,the Chongqing Branch of the National Clinical Research Center for Ocular Diseases,the Chongqing Key Laboratory of Ophthalmology(CSTC,2008CA5003)the Program for Youth Innovation in Future Medicine,Chongqing Medical University(w0047).
文摘Vogt–Koyanagi–Harada(VKH)disease is a leading cause of blindness in young and middle-aged people.However,the etiology of VKH disease remains unclear.Here,we performed the first trio-based whole-exome sequencing study,which enrolled 25 VKH patients and 50 controls,followed by a study of 2081 VKH patients from a Han Chinese population to uncover detrimental mutations.A total of 15 de novo mutations in VKH patients were identified,with one of the most important being the membrane palmitoylated protein 2(MPP2)p.K315N(MPP2-N315)mutation.The MPP2-N315 mutation was highly deleterious according to bioinformatic predictions.Additionally,this mutation appears rare,being absent from the 1000 Genome Project and Genome Aggregation Database,and it is highly conserved in 10 species,including humans and mice.Subsequent studies showed that pathological phenotypes and retinal vascular leakage were aggravated in MPP2-N315 mutation knock-in or MPP2-N315 adeno-associated virus-treated mice with experimental autoimmune uveitis(EAU).In vitro,we used clustered regularly interspaced short palindromic repeats(CRISPR‒Cas9)gene editing technology to delete intrinsic MPP2 before overexpressing wild-type MPP2 or MPP2-N315.Levels of cytokines,such as IL-1β,IL-17E,and vascular endothelial growth factor A,were increased,and barrier function was destroyed in the MPP2-N315 mutant ARPE19 cells.Mechanistically,the MPP2-N315 mutation had a stronger ability to directly bind to ANXA2 than MPP2-K315,as shown by LC‒MS/MS and Co-IP,and resulted in activation of the ERK3/IL-17E pathway.Overall,our results demonstrated that the MPP2-K315N mutation may increase susceptibility to VKH disease.
基金supported by the Natural Science Foundation of China(81670893,82121003)Science and Technology Department of Sichuan Province(2021JDZH0031)Chinese Academy of Medical Sciences(2019-I2M-5-032)。
文摘Mutations in serologically defined colon cancer autoantigen protein 8(SDCCAG8)were first identified in retinal ciliopathy families a decade ago with unknown function.To investigate the pathogenesis of SDCCAG8-associated retinal ciliopathies in vivo,we employed CRISPR/Cas9-mediated homology-directed recombination(HDR)to generate two knock-in mouse models,Sdccag8^(Y236X/Y236X) and Sdccag8^(E451GfsX467/E451GfsX467),which carry truncating mutations of the mouse Sdccag8,corresponding to mutations that cause Bardet-Biedl syndrome(BBS)and Senior-L?ken syndrome(SLS)(c.696T>G p.Y232X and c.1339-1340ins G p.E447GfsX463)in humans,respectively.The two mutant Sdccag8 knock-in mice faithfully recapitulated human SDCCAG8-associated BBS phenotypes such as rod-cone dystrophy,cystic renal disorder,polydactyly,infertility,and growth retardation,with varied age of onset and severity depending on the hypomorphic strength of the Sdccag8 mutations.To the best of our knowledge,these knock-in mouse lines are the first BBS mouse models to present with the polydactyly phenotype.Major phototransduction protein mislocalization was also observed outside the outer segment after initiation of photoreceptor degeneration.Impaired cilia were observed in the mutant photoreceptors,renal epithelial cells,and mouse embryonic fibroblasts derived from the knock-in mouse embryos,suggesting that SDCCAG8 plays an essential role in ciliogenesis,and cilium defects are a primary driving force of SDCCAG8-associated retinal ciliopathies.
基金supported by the National Natural Science Foundation of China(No.81790643,81970839,82271105,and 82121003)the Sichuan Science and Technology Program(China)(No.2021YFS0033,2021YFS0404,2021YFS0369,and 2020ZYD037)the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(China)(No.2019-I2M-5-032).
文摘The retinal pigment epithelium(RPE)and choroid are located behind the human retina and have multiple functions in the human visual system.Knowledge of the RPE and choroid cells and their gene expression profiles are fundamental for understanding retinal disease mechanisms and therapeutic strategies.Here,we sequenced the RNA of about 0.3 million single cells from human RPE and choroids across two regions and seven ages,revealing regional and age differences within the human RPE and choroid.Cell–cell interactions highlight the broad connectivity networks between the RPE and different choroid cell types.Moreover,the transcription factors and their target genes change during aging.The coding of somatic variations increases during aging in the human RPE and choroid at the single-cell level.Moreover,we identified ELN as a candidate for improving RPE degeneration and choroidal structure during aging.The mapping of the molecular architecture of the human RPE and choroid improves our understanding of the human vision support system and offers potential insights into the intervention targets for retinal diseases.
基金supported by the National Natural Science Foundation of China(No.82101153,82000913,81970841,82121003,and 82071009)the Sichuan Science and Technology Program,China(No.2022YFS0598,2021YFS0386,2021YFS0369,and 2021JDGD0036)+4 种基金the CAMS Innovation Fund for Medical Sciences,China(No.2019-12M-5-032)the Department of Science and Technology of Qinghai Province,China(No.2022-HZ-814)the fund for Sichuan Provincial People's Hospital,China(No.2021QN01)the Department of Chengdu Science and Technology,China(No.2021-YF05-01316-SN)the Huanhua Outstanding Scholar Program for Sichuan Provincial People's Hospital(China)to Xianjun Zhu.The funders had no role in the study design,data collection,analysis,or preparation of the manuscript.
文摘Endoplasmic reticulum(ER)membrane protein complex(EMC)is required for the co-translational insertion of newly synthesized multi-transmembrane proteins.Compromised EMC function in different cell types has been implicated in multiple diseases.Using inducible genetic mouse models,we revealed defects in retinal vascularization upon endothelial cell(EC)specific deletion of Emc1,the largest subunit of EMC.Loss of Emc1 in ECs led to reduced vascular progression and vascular density,diminished tip cell sprouts,and vascular leakage.We then performed an unbiased transcriptomic analysis on human retinal microvascular endothelial cells(HRECs)and revealed a pivotal role of EMC1 in theβ-catenin signaling pathway.Further in-vitro and in-vivo experiments proved that loss of EMC1 led to compromisedβ-catenin signaling activity through reduced expression of Wnt receptor FZD4,which could be restored by lithium chloride(LiCl)treatment.Driven by these findings,we screened genomic DNA samples from familial exudative vitreoretinopathy(FEVR)patients and identified one heterozygous variant in EMC1 that co-segregated with FEVR phenotype in the family.In-vitro expression experiments revealed that this variant allele failed to facilitate the expression of FZD4 on the plasma membrane and activate theβ-catenin signaling pathway,which might be a main cause of FEVR.In conclusion,our findings reveal that variants in EMC1 gene cause compromisedβ-catenin signaling activity,which may be associated with the pathogenesis of FEVR.
基金National Key Research and Development Plan(No.2017YFC0113901)the Department of Science and Technology of Sichuan Province,China(No.2017JZ0039)+1 种基金the Science Research Project for Cadres’Health Care of Sichuan Province(No.2017-205)Science and Technology Department of Sichuan Province(No.2019JDJQ0031,No.2020ZYD035)。
文摘AIM:To systematically understand the genetic association between methylenetetrahydrofolate reductase(MTHFR)C677 T polymorphism and primary open angle glaucoma(POAG).METHODS:A comprehensive literature search in Google Scholar,Pub Med,Science Citation Index,Foreign Medical Literature Retrieval Service,Chinese National Knowledge Infrastructure and Wanfang Databases was performed to collect all eligible studies up to August 2019.Study selection,data abstraction and study quality evaluation were performed by two independent investigators.Odds ratios(ORs)with 95%confidence intervals(CIs)were calculated to assess the association.RESULTS:Eighteen case-control studies including 2156 cases and 2201 controls were identified.There was no significant difference in the terms of MTHFR C677 T polymorphism and POAG in the Caucasian population(for T vs C OR=1.11,95%CI:0.88 to 1.39;for TT vs CC OR=1.01,95%CI:0.76 to 1.36;for TT+TC vs CC OR=1.15,95%CI:0.84 to 1.58 and for TT vs TC+CC OR=1.02,95%CI:0.78 to 1.33).However,a significant effect was revealed in the Asian population(for T vs C OR=1.34,95%CI:1.12 to 1.59;for TT+TC vs CC OR=1.41,95%CI:1.14 to 1.76).CONCLUSION:Based on 18 eligible studies,we provide a correlation between MTHFR C677 T polymorphism and POAG among the Asians subgroup indicating that the T allele or TT+TC genotype may play a critical role in POAG development in Asians.
基金We thank all the patients and their families for participating in this study.This research project was supported by the National Precision Medicine Project(2016YFC0905200)the National Natural Science Foundation of China(81790643,81970839,81871722,82121003 and 82072373)+2 种基金the Sichuan Science and Technology Program(2020YFS0014,2020YFS0558,2020ZYD035,2020ZYD037,2021ZYD0082,2019JDJQ0031,2021YFS0369,2021JDGD0036,2021YFS0033,2021YFS0404,2021YFS0388,2022JDTD0024 and 2020JDTD0028)the Grant from Chinese Academy of Medical Sciences(No.2019-I2M5-032)Sichuan Provincial Health Commission(20ZDCX002).
文摘Critical coronavirus disease 2019(COVID-19)is associated with high mortality and potential genetic factors have been reported to be involved in the development of critical COVID-19.We performed a genome-wide association study to identify the genetic factors responsible for developing critical COVID-19.632 critical patients with COVID-19 and 3021 healthy controls from the Chinese population were recruited.First,we identified a genome-wide significant difference of IL-6 rs2069837(p=9.73×10^(−15),OR=0.41)between 437 critical patients with COVID-19 and 2551 normal controls in the discovery cohort.
基金supported by the National Natural Science Foundation of China(81970841,82101160,82121003)the Department of Science and Technology of Sichuan Province(2023ZYD0172,2023YFS0161)+3 种基金the program of Science and Technology International Cooperation Project of Qinghai province(China)(No.2022-HZ-814)Sichuan Intellectual Property Office(China)(No.2022-ZS-0070)the CAMS Innovation Fund for Medical Sciences(2019-12M-5-032)Open Project of Henan Provincial Key Laboratory of Ophthalmology and Visual Science(20KFKT02).
文摘Inherited retinal dystrophies (IRDs) are major causes of visual impairment and irreversible blindness worldwide, while the precise molecular and genetic mechanisms are still elusive. N6-methyladenosine (m^(6)A) modification is the most prevalent internal modification in eukaryotic mRNA. YTH domain containing 2 (YTHDC2), an m^(6)A reader protein, has recently been identified as a key player in germline development and human cancer. However, its contribution to retinal function remains unknown. Here, we explore the role of YTHDC2 in the visual function of retinal rod photoreceptors by generating rod-specific Ythdc2 knockout mice. Results show that Ythdc2 deficiency in rods causes diminished scotopic ERG responses and progressive retinal degeneration. Multi-omics analysis further identifies Ppef2 and Pde6b as the potential targets of YTHDC2 in the retina. Specifically, via its YTH domain, YTHDC2 recognizes and binds m^(6)A-modified Ppef2 mRNA at the coding sequence and Pde6b mRNA at the 5′-UTR, resulting in enhanced translation efficiency without affecting mRNA levels. Compromised translation efficiency of Ppef2 and Pde6b after YTHDC2 depletion ultimately leads to decreased protein levels in the retina, impaired retinal function, and progressive rod death. Collectively, our finding highlights the importance of YTHDC2 in visual function and photoreceptor survival, which provides an unreported elucidation of IRD pathogenesis via epitranscriptomics.
