Idiopathic pulmonary fibrosis(IPF)is a progressive pulmonary disease that leads to interstitial inflammation,lung damage,and eventually life-threatening complications.Among various pathologic factors,Smad4 is a pivota...Idiopathic pulmonary fibrosis(IPF)is a progressive pulmonary disease that leads to interstitial inflammation,lung damage,and eventually life-threatening complications.Among various pathologic factors,Smad4 is a pivotal molecule involved in the progression and exacerbation of IPF.It mediates nuclear transfer of Smad2/Smad3 complexes and initiates the transcription of fibrosis-promoting genes.Thus,the inhibition of Smad4 expression in pulmonary fibroblasts by small interfering RNAs(siRNAs)might be a promising therapeutic strategy for IPF.Herein,we engineered exosome membranes(EM)by cationic lipid(i.e.,DOTAP)to load siRNAs against Smad4(DOTAP/siSmad4@EM),and investigated their specific delivery to pulmonary fibroblasts for treating IPF in a mouse model via pulmonary administration.As reference nanoscaffolds,undecorated DOTAP/siSmad4 complexes(lipoplexes,consisting of cationic lipid DOTAP and siRNAs)and siSmad4-loaded lipid nanoparticles(DOTAP/siSmad4@lipo,consisting of lipoplexes fused with DPPC–Chol liposomes)were also prepared.The results showed that DOTAP/siSmad4@EM exhibited a higher cellular uptake and gene silencing efficacies in mouse pulmonary fibroblasts(viz.,MLg2908)as compared to the two reference nanoscaffolds.Furthermore,the outcomes of the in vivo experiments illustrated that DOTAP/siSmad4@EM could significantly down-regulate the Smad4 expression with augmented anti-fibrosis efficiency.Additionally,the DOTAP/siSmad4@EM conferred excellent biocompatibility with low cytokine levels in bronchoalveolar lavage fluid and proinflammatory responses in the pulmonary area.Taken together,the outcomes of our investigation imply that specific inhibition of Smad4 expression in pulmonary fibroblasts by pulmonary administrated DOTAP/siSmad4@EM is a promising therapeutic strategy for IPF,which could safely and effectively deliver siRNA drugs to the targeted site of action.展开更多
The sweet potato weevil(Cylas formicarius)is a critical pest producing enor-mous global losses in sweet potato crops.Traditional pest management approaches for sweet potato weevil,primarily using chemical pesticides,c...The sweet potato weevil(Cylas formicarius)is a critical pest producing enor-mous global losses in sweet potato crops.Traditional pest management approaches for sweet potato weevil,primarily using chemical pesticides,causes pollution,food safety is-sues,and harming natural enemies.While RNA interference(RNAi)is a promising envi-ronmentally friendly approach to pest control,its efficacy in controlling the sweet potato weevil has not been extensively studied.In this study,we selected a potential target for controlling C.formicarius,the Troponin I gene(wupA),which is essential for muscula-ture composition and crucial for fundamental life activities.We determined that wupA is abundantly expressed throughout all developmental stages of the sweet potato weevil.We evaluated the efficiency of double-stranded RNAs in silencing the wupA gene via microinjection and oral feeding of sweet potato weevil larvae at different ages.Our find-ings demonstrate that both approaches significantly reduced the expression of wupA and produced high mortality.Moreover,the 1st instar larvae administered dswupA exhibited significant growth inhibition.We assessed the toxicity of dswupA on the no-target insect silkworm and assessed its safety.Our study indicates that wupA knockdown can inhibit the growth and development of C.formicarius and offer a potential target gene for envi-ronmentally friendly control.展开更多
Autoreactive CD8^(+)T cells play a key role in type 1 diabetes(T1D),but the antigen spectrum that activates autoreactive CD8^(+)T cells remains unclear.Endoplasmic reticulum stress(ERS)has been implicated inβ-cell au...Autoreactive CD8^(+)T cells play a key role in type 1 diabetes(T1D),but the antigen spectrum that activates autoreactive CD8^(+)T cells remains unclear.Endoplasmic reticulum stress(ERS)has been implicated inβ-cell autoantigen generation.Here,we analyzed the major histocompatibility complex class I(MHC-I)-associated immunopeptidome(MIP)of isletβ-cells under steady and ERS conditions and found that ERS reshaped the MIP ofβ-cells and promoted the MHC-I presentation of a panel of conventional self-peptides.Among them,OTUB2_(58-66 ) showed immunodominance,and the corresponding autoreactive CD8^(+)T cells were diabetogenic in nonobese diabetic(NOD)mice.High glucose intake upregulated pancreatic OTUB2 expression and amplified the OTUB2_(58-66 )-specific CD8^(+)T-cell response in NOD mice.Repeated OTUB2_(58-66 )administration significantly reduced the incidence of T1D in NOD mice.Interestingly,peripheral blood mononuclear cells(PBMCs)from patients with T1D,but not from healthy controls,showed a positive IFN-γresponse to human OTUB2 peptides.This study provides not only a new explanation for the role of ERS in promotingβ-cell-targeted autoimmunity but also a potential target for the prevention and treatment of T1D.The data are available via ProteomeXchange with the identifier PXD041227.展开更多
Autoreactive CD8^(+)T cells,which play an indispensable role inβcell destruction,represent an emerging target for the prevention of type 1 diabetes(T1D).Altered peptide ligands(APLs)can efficiently induce antigen-spe...Autoreactive CD8^(+)T cells,which play an indispensable role inβcell destruction,represent an emerging target for the prevention of type 1 diabetes(T1D).Altered peptide ligands(APLs)can efficiently induce antigen-specific T cells anergy,apoptosis or shifts in the immune response.Here,we found that HLA-A*0201-restricted CD8^(+)T cell responses against a primaryβ-cell autoantigen insulin epitope InsB15–14 were present in both NOD.β2m null.HHD NOD mice and T1D patients.We generated several APL candidates for InsB15–14 by residue substitution at the p6 position.Only H6F exhibited an inhibitory effect on mInsB1_(5–14)-specific CD8^(+)T cell responses in vitro.H6F treatment significantly reduced the T1D incidence,which was accompanied by diminished autoreactive CD8^(+)T cell responses to mInsB15-14,inhibited infiltration of CD8^(+)and CD4^(+)T cells in the pancreas and reduced pro-inflammatory cytokine production in pancreatic and splenic T cells in NOD.β2m^(null).HHD mice.Mechanistically,H6F treatment significantly augmented a tiny portion of CD8^(+)CD25^(+)Foxp3^(+)T cells in the spleen and especially in the pancreas.This subset exhibited typical Treg phenotypes and required peptide-specific restimulation to exert immunosuppressive activity.Therefore,this APL H6F may be a promising candidate with potential clinical application value for antigen-specific prevention of T1D.展开更多
基金supported by the Liaoning Pan Deng Xue Zhe Scholar(grant No.XLYC2002061)the National Natural Science Foundation of China(grant No.82173768)+7 种基金the Overseas Expertise Introduction Project for Discipline Innovation(“111 Project”)(grant No.D20029)M.Y.thanks Independent Research Fund Denmark for the financial support(Grant ID:10.46540/3105-00249B)L.W.acknowledges the financial supports of National Natural Science Foundation of China(grant No.82204316)China Postdoctoral Science Foundation(grant Nos.2021TQ0219 and 2022MD713776)H.B.thanks the financial support from National Natural Science Foundation of China,Govt.of China(grant No.82050410448)Fellowship of China Postdoctoral Science Foundation,Govt.of China(grant No.2021MD703857)D.C.acknowledges financial support from Nature Science Foundation of Liaoning Province(grant No.2022-MS-241)Ministry of Education Chunhui Program(2020).
文摘Idiopathic pulmonary fibrosis(IPF)is a progressive pulmonary disease that leads to interstitial inflammation,lung damage,and eventually life-threatening complications.Among various pathologic factors,Smad4 is a pivotal molecule involved in the progression and exacerbation of IPF.It mediates nuclear transfer of Smad2/Smad3 complexes and initiates the transcription of fibrosis-promoting genes.Thus,the inhibition of Smad4 expression in pulmonary fibroblasts by small interfering RNAs(siRNAs)might be a promising therapeutic strategy for IPF.Herein,we engineered exosome membranes(EM)by cationic lipid(i.e.,DOTAP)to load siRNAs against Smad4(DOTAP/siSmad4@EM),and investigated their specific delivery to pulmonary fibroblasts for treating IPF in a mouse model via pulmonary administration.As reference nanoscaffolds,undecorated DOTAP/siSmad4 complexes(lipoplexes,consisting of cationic lipid DOTAP and siRNAs)and siSmad4-loaded lipid nanoparticles(DOTAP/siSmad4@lipo,consisting of lipoplexes fused with DPPC–Chol liposomes)were also prepared.The results showed that DOTAP/siSmad4@EM exhibited a higher cellular uptake and gene silencing efficacies in mouse pulmonary fibroblasts(viz.,MLg2908)as compared to the two reference nanoscaffolds.Furthermore,the outcomes of the in vivo experiments illustrated that DOTAP/siSmad4@EM could significantly down-regulate the Smad4 expression with augmented anti-fibrosis efficiency.Additionally,the DOTAP/siSmad4@EM conferred excellent biocompatibility with low cytokine levels in bronchoalveolar lavage fluid and proinflammatory responses in the pulmonary area.Taken together,the outcomes of our investigation imply that specific inhibition of Smad4 expression in pulmonary fibroblasts by pulmonary administrated DOTAP/siSmad4@EM is a promising therapeutic strategy for IPF,which could safely and effectively deliver siRNA drugs to the targeted site of action.
基金supported from the National Key R&D Program of China(2022YFE0116500)the Shen zhen Science and Technology Program(Grant No.KQTD20180411143628272)and Agricultural Science and Technology Innovation Program(ASTIP).
文摘The sweet potato weevil(Cylas formicarius)is a critical pest producing enor-mous global losses in sweet potato crops.Traditional pest management approaches for sweet potato weevil,primarily using chemical pesticides,causes pollution,food safety is-sues,and harming natural enemies.While RNA interference(RNAi)is a promising envi-ronmentally friendly approach to pest control,its efficacy in controlling the sweet potato weevil has not been extensively studied.In this study,we selected a potential target for controlling C.formicarius,the Troponin I gene(wupA),which is essential for muscula-ture composition and crucial for fundamental life activities.We determined that wupA is abundantly expressed throughout all developmental stages of the sweet potato weevil.We evaluated the efficiency of double-stranded RNAs in silencing the wupA gene via microinjection and oral feeding of sweet potato weevil larvae at different ages.Our find-ings demonstrate that both approaches significantly reduced the expression of wupA and produced high mortality.Moreover,the 1st instar larvae administered dswupA exhibited significant growth inhibition.We assessed the toxicity of dswupA on the no-target insect silkworm and assessed its safety.Our study indicates that wupA knockdown can inhibit the growth and development of C.formicarius and offer a potential target gene for envi-ronmentally friendly control.
基金National Natural Science Foundation of China(No.82071825 and No.81871301)National Key Research and Development Program(No.2016YFA0502204)Shandong Provincial Natural Science Fund(ZR2023MH201).
文摘Autoreactive CD8^(+)T cells play a key role in type 1 diabetes(T1D),but the antigen spectrum that activates autoreactive CD8^(+)T cells remains unclear.Endoplasmic reticulum stress(ERS)has been implicated inβ-cell autoantigen generation.Here,we analyzed the major histocompatibility complex class I(MHC-I)-associated immunopeptidome(MIP)of isletβ-cells under steady and ERS conditions and found that ERS reshaped the MIP ofβ-cells and promoted the MHC-I presentation of a panel of conventional self-peptides.Among them,OTUB2_(58-66 ) showed immunodominance,and the corresponding autoreactive CD8^(+)T cells were diabetogenic in nonobese diabetic(NOD)mice.High glucose intake upregulated pancreatic OTUB2 expression and amplified the OTUB2_(58-66 )-specific CD8^(+)T-cell response in NOD mice.Repeated OTUB2_(58-66 )administration significantly reduced the incidence of T1D in NOD mice.Interestingly,peripheral blood mononuclear cells(PBMCs)from patients with T1D,but not from healthy controls,showed a positive IFN-γresponse to human OTUB2 peptides.This study provides not only a new explanation for the role of ERS in promotingβ-cell-targeted autoimmunity but also a potential target for the prevention and treatment of T1D.The data are available via ProteomeXchange with the identifier PXD041227.
基金supported by the National Natural Science Foundation of China(No.31570931 and No.31771002)the National Key Project for Research&Development of China(Grant no.2016YFA0502204).
文摘Autoreactive CD8^(+)T cells,which play an indispensable role inβcell destruction,represent an emerging target for the prevention of type 1 diabetes(T1D).Altered peptide ligands(APLs)can efficiently induce antigen-specific T cells anergy,apoptosis or shifts in the immune response.Here,we found that HLA-A*0201-restricted CD8^(+)T cell responses against a primaryβ-cell autoantigen insulin epitope InsB15–14 were present in both NOD.β2m null.HHD NOD mice and T1D patients.We generated several APL candidates for InsB15–14 by residue substitution at the p6 position.Only H6F exhibited an inhibitory effect on mInsB1_(5–14)-specific CD8^(+)T cell responses in vitro.H6F treatment significantly reduced the T1D incidence,which was accompanied by diminished autoreactive CD8^(+)T cell responses to mInsB15-14,inhibited infiltration of CD8^(+)and CD4^(+)T cells in the pancreas and reduced pro-inflammatory cytokine production in pancreatic and splenic T cells in NOD.β2m^(null).HHD mice.Mechanistically,H6F treatment significantly augmented a tiny portion of CD8^(+)CD25^(+)Foxp3^(+)T cells in the spleen and especially in the pancreas.This subset exhibited typical Treg phenotypes and required peptide-specific restimulation to exert immunosuppressive activity.Therefore,this APL H6F may be a promising candidate with potential clinical application value for antigen-specific prevention of T1D.
