Retinitis pigmentosa(RP)is a form of inherited retinal degenerative diseases that ultimately involves the macula,which is present in primates but not in the rodents.Therefore,creating nonhuman primate(NHP)models of RP...Retinitis pigmentosa(RP)is a form of inherited retinal degenerative diseases that ultimately involves the macula,which is present in primates but not in the rodents.Therefore,creating nonhuman primate(NHP)models of RP is of critical importance to study its mechanism of pathogenesis and to evaluate potential therapeutic options in the future.Here we applied adeno-associated virus(AAV)-delivered CRISPR/SaCas9 technology to knockout the RHO gene in the retinae of the adult rhesus macaque(Macaca mulatta)to investigate the hypothesis whether non-germline mutation of the RHO gene is sufficient to recapitulate RP.Through a series of studies,we were able to demonstrate successful somatic editing of the RHO gene and reduced RHO protein expression.More importantly,the mutant macaque retinae displayed clinical RP phenotypes,including photoreceptor degeneration,retinal thinning,abnormal rod subcellular structures,and reduced photoresponse.Therefore,we suggest somatic editing of the RHO gene is able to phenocopy RP,and the reduced time span in generating NHP mutant accelerates RP research and expands the utility of NHP model for human disease study.展开更多
Antiangiogenesis gene therapy based on adeno-associated virus(AAV)vectors represents a promising advancement in the treatment of neovascular age-related macular degeneration(nAMD),providing an alternative to antibody-...Antiangiogenesis gene therapy based on adeno-associated virus(AAV)vectors represents a promising advancement in the treatment of neovascular age-related macular degeneration(nAMD),providing an alternative to antibody-based therapies.However,the development of a safe and effective AAV vector capable of precisely targeting neovascularization and choroidal leakage remains a critical unmet need.In the present study,we engineered a novel intravitreally administered AAV vector with retinal-pigment-epithelium(RPE)-specific tropism.This vector demonstrated robust and localized gene expression in RPE cells while maintaining a favorable safety profile.The RPE-tropic AAV vector delivered a dual-acting antibody against vascular endothelial growth factor(VEGF)and angiopoietin-2(ANG-2),exhibiting strong therapeutic efficacy and tolerability in both rodent and nonhuman primate choroidal neovascularization models.Based on the promising preclinical data,a single-center,single-arm,investigator-initiated trial(ChiCTR2400085329)was conducted to assess its safety and efficacy in patients with nAMD.The RPE-tropic AAV vector expressing anti-VEGF-A and anti-ANG-2 effectively alleviated disease progression and was well tolerated in the clinical setting.These findings highlight the potential of this engineered AAV-RPE capsid as a versatile platform for gene therapy,not only for nAMD but also for other ocular diseases involving RPE cells.展开更多
基金the National Key Research and Development Program of China(2020YFA0112200,2016YFA0400900,and 2018YFA0801403)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16020603,XDB39000000,and XDB32060200)+3 种基金the National Natural Science Foundation of China(81925009,81790644,61890953,31322024,81371066,91432104,81900855,31900712,and 31800901)Guangdong Provincial Key Research and Development Program(2019B030335001 and 2018B030338001)Anhui Provincial Natural Science Foundation(1808085MH289 and 1908085MC66)the Fundamental Research Funds for the Central Universities(WK2070000174 and WK2090050048)。
文摘Retinitis pigmentosa(RP)is a form of inherited retinal degenerative diseases that ultimately involves the macula,which is present in primates but not in the rodents.Therefore,creating nonhuman primate(NHP)models of RP is of critical importance to study its mechanism of pathogenesis and to evaluate potential therapeutic options in the future.Here we applied adeno-associated virus(AAV)-delivered CRISPR/SaCas9 technology to knockout the RHO gene in the retinae of the adult rhesus macaque(Macaca mulatta)to investigate the hypothesis whether non-germline mutation of the RHO gene is sufficient to recapitulate RP.Through a series of studies,we were able to demonstrate successful somatic editing of the RHO gene and reduced RHO protein expression.More importantly,the mutant macaque retinae displayed clinical RP phenotypes,including photoreceptor degeneration,retinal thinning,abnormal rod subcellular structures,and reduced photoresponse.Therefore,we suggest somatic editing of the RHO gene is able to phenocopy RP,and the reduced time span in generating NHP mutant accelerates RP research and expands the utility of NHP model for human disease study.
基金supported by the National Key Basic Research Program of China(grant no.2020YFA0112200)the National Natural Science Foundation of China(grant nos.81925009 and 82021001)+3 种基金the CAS Project for Young Scientists in Basic Research(grant no.YSBR-013)the National Key R&D program of China(grant no.2023YFC3403400)the Innovative Research Team of High-Level Local Universities in Shanghai(grant no.SHSMU-ZDCX20211100)the Anhui Provincial Key Research and Development Project(grant no.2023S07020009).
文摘Antiangiogenesis gene therapy based on adeno-associated virus(AAV)vectors represents a promising advancement in the treatment of neovascular age-related macular degeneration(nAMD),providing an alternative to antibody-based therapies.However,the development of a safe and effective AAV vector capable of precisely targeting neovascularization and choroidal leakage remains a critical unmet need.In the present study,we engineered a novel intravitreally administered AAV vector with retinal-pigment-epithelium(RPE)-specific tropism.This vector demonstrated robust and localized gene expression in RPE cells while maintaining a favorable safety profile.The RPE-tropic AAV vector delivered a dual-acting antibody against vascular endothelial growth factor(VEGF)and angiopoietin-2(ANG-2),exhibiting strong therapeutic efficacy and tolerability in both rodent and nonhuman primate choroidal neovascularization models.Based on the promising preclinical data,a single-center,single-arm,investigator-initiated trial(ChiCTR2400085329)was conducted to assess its safety and efficacy in patients with nAMD.The RPE-tropic AAV vector expressing anti-VEGF-A and anti-ANG-2 effectively alleviated disease progression and was well tolerated in the clinical setting.These findings highlight the potential of this engineered AAV-RPE capsid as a versatile platform for gene therapy,not only for nAMD but also for other ocular diseases involving RPE cells.
