Molecular Therapy. Methods & Clinical Development最新文献_第7页

Good reasons for targeting SARS-CoV-2 by engineered extracellular vesicles. 利用工程细胞外囊泡靶向SARS-CoV-2的充分理由。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-03-09 eCollection Date: 2022-06-09 DOI: 10.1016/j.omtm.2022.02.003

Ario de Marco, Lucio Barile

引用次数: 0

Follistatin-like 1 promotes proliferation of matured human hypoxic iPSC-cardiomyocytes and is secreted by cardiac fibroblasts. 卵泡抑素样1促进成熟的人缺氧ipsc心肌细胞的增殖，并由心脏成纤维细胞分泌。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-02-23 eCollection Date: 2022-06-09 DOI: 10.1016/j.omtm.2022.02.005

Marijn C Peters, Sofia Di Martino, Thomas Boelens, Jiabin Qin, Alain van Mil, Pieter A Doevendans, Steven A J Chamuleau, Joost P G Sluijter, Klaus Neef

{"title":"Follistatin-like 1 promotes proliferation of matured human hypoxic iPSC-cardiomyocytes and is secreted by cardiac fibroblasts.","authors":"Marijn C Peters, Sofia Di Martino, Thomas Boelens, Jiabin Qin, Alain van Mil, Pieter A Doevendans, Steven A J Chamuleau, Joost P G Sluijter, Klaus Neef","doi":"10.1016/j.omtm.2022.02.005","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.02.005","url":null,"abstract":"The human heart has limited regenerative capacity. Therefore, patients often progress to heart failure after ischemic injury, despite advances in reperfusion therapies generally decreasing mortality. Depending on its glycosylation state, Follistatin-like 1 (FSTL1) has been shown to increase cardiomyocyte (CM) proliferation, decrease CM apoptosis, and prevent cardiac rupture in animal models of ischemic heart disease. To explore its therapeutic potential, we used a human in vitro model of cardiac ischemic injury with human induced pluripotent stem cell-derived CMs (iPSC-CMs) and assessed regenerative effects of two differently glycosylated variants of human FSTL1. Furthermore, we investigated the FSTL1-mediated interplay between human cardiac fibroblasts (cFBs) and iPSC-CMs in hypoxia. Both FSTL1 variants increased viability, while only hypo-glycosylated FSTL1 increased CM proliferation post-hypoxia. Human fetal cardiac fibroblasts (fcFBs) expressed and secreted FSTL1 under normoxic conditions, while FSTL1 secretion increased by iPSC-cFBs upon hypoxia but decreased in iPSC-CMs. Co-culture of iPSC-CMs and cFBs increased FSTL1 secretion compared with cFB mono-culture. Taken together, we confirm that FSTL1 induces iPSC-CM proliferation in a human cardiac in vitro hypoxia damage model. Furthermore, we show hypoxia-related FSTL1 secretion by human cFBs and indications for FSTL1-mediated intercellular communication between cardiac cell types in response to hypoxic conditions.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"3-16"},"PeriodicalIF":4.7,"publicationDate":"2022-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8917270/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40311605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

In vivo targeting of a variant causing vanishing white matter using CRISPR/Cas9. 利用CRISPR/Cas9在体内靶向一种导致白质消失的变体。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-02-23 eCollection Date: 2022-06-09 DOI: 10.1016/j.omtm.2022.02.006

Anne E J Hillen, Martina Hruzova, Tanja Rothgangl, Marjolein Breur, Marianna Bugiani, Marjo S van der Knaap, Gerald Schwank, Vivi M Heine

{"title":"In vivo targeting of a variant causing vanishing white matter using CRISPR/Cas9.","authors":"Anne E J Hillen, Martina Hruzova, Tanja Rothgangl, Marjolein Breur, Marianna Bugiani, Marjo S van der Knaap, Gerald Schwank, Vivi M Heine","doi":"10.1016/j.omtm.2022.02.006","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.02.006","url":null,"abstract":"Vanishing white matter (VWM) is a leukodystrophy caused by recessive variants in subunits of eIF2B. At present, no curative treatment is available and patients often die at young age. Due to its monogenic nature, VWM is a promising candidate for the development of CRISPR/Cas9-mediated gene therapy. Here we tested a dual-AAV approach in VWM mice encoding CRISPR/Cas9 and a DNA donor template to correct a pathogenic variant in Eif2b5. We performed sequencing analysis to assess gene correction rates and examined effects on the VWM phenotype, including motor behavior. Sequence analysis demonstrated that over 90% of CRISPR/Cas9-induced edits at the targeted locus are insertion or deletion (indel) mutations, rather than precise corrections from the DNA donor template by homology-directed repair. Around half of the CRISPR/Cas9-treated animals died prematurely. VWM mice showed no improvement in motor skills, weight, or neurological scores at 7 months of age, and CRISPR/Cas9-treated controls displayed an induced VWM phenotype. In conclusion, CRISPR/Cas9-induced DNA double-strand breaks (DSBs) at the Eif2b5 locus did not lead to sufficient correction of the VWM variant. Moreover, indel formation in Eif2b5 induced an exacerbated VWM phenotype. Therefore, DSB-independent strategies like base- or prime editing might better suited for VWM correction.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"17-25"},"PeriodicalIF":4.7,"publicationDate":"2022-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8917273/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40311604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Engineering new metabolic pathways in isolated cells for the degradation of guanidinoacetic acid and simultaneous production of creatine. 在分离细胞中设计新的代谢途径以降解胍基乙酸并同时产生肌酸。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-02-22 eCollection Date: 2022-06-09 DOI: 10.1016/j.omtm.2022.02.007

Marzia Bianchi, Luigia Rossi, Francesca Pierigè, Pietro De Angeli, Mattia Paolo Aliano, Claudia Carducci, Emanuele Di Carlo, Tiziana Pascucci, Francesca Nardecchia, Vincenzo Leuzzi, Mauro Magnani

{"title":"Engineering new metabolic pathways in isolated cells for the degradation of guanidinoacetic acid and simultaneous production of creatine.","authors":"Marzia Bianchi, Luigia Rossi, Francesca Pierigè, Pietro De Angeli, Mattia Paolo Aliano, Claudia Carducci, Emanuele Di Carlo, Tiziana Pascucci, Francesca Nardecchia, Vincenzo Leuzzi, Mauro Magnani","doi":"10.1016/j.omtm.2022.02.007","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.02.007","url":null,"abstract":"Here we report, for the first time, the engineering of human red blood cells (RBCs) with an entire metabolic pathway as a potential strategy to treat patients with guanidinoacetate methyltransferase (GAMT) deficiency, capable of reducing the high toxic levels of guanidinoacetate acid (GAA) and restoring proper creatine levels in blood and tissues. We first produced a recombinant form of native human GAMT without any tags to encapsulate into RBCs. Due to the poor solubility and stability features of the recombinant enzyme, both bioinformatics studies and extensive optimization work were performed to select a mutant GAMT enzyme, where only four critical residues were replaced, as a lead candidate. However, GAMT-loaded RBCs were ineffective in GAA consumption and creatine production because of the limiting intra-erythrocytic S-adenosyl methionine (SAM) content unable to support GAMT activity. Therefore, a recombinant form of human methionine adenosyl transferase (MAT) was developed. RBCs co-entrapped with both GAMT and MAT enzymes performed, in vitro, as a competent cellular bioreactor to remove GAA and produce creatine, fueled by physiological concentrations of methionine and the ATP generated by glycolysis. Our results highlight that metabolic engineering of RBCs is possible and represents proof of concept for the design of novel therapeutic approaches.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"26-40"},"PeriodicalIF":4.7,"publicationDate":"2022-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8917272/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40311606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Erratum: Monitoring CAR T cell generation with a CD8-targeted lentiviral vector by single-cell transcriptomics. 勘误:通过单细胞转录组学监测cd8靶向慢病毒载体的CAR - T细胞生成。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-01-24 eCollection Date: 2022-03-10 DOI: 10.1016/j.omtm.2022.01.010

Filippos T Charitidis, Elham Adabi, Frederic B Thalheimer, Colin Clarke, Christian J Buchholz

引用次数: 3

Prednisolone reduces the interferon response to AAV in cynomolgus macaques and may increase liver gene expression. 强的松龙降低食蟹猴对AAV的干扰素反应，并可能增加肝脏基因表达。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-01-19 eCollection Date: 2022-03-10 DOI: 10.1016/j.omtm.2022.01.007

Lili Wang, Claude C Warzecha, Alexander Kistner, Jessica A Chichester, Peter Bell, Elizabeth L Buza, Zhenning He, M Betina Pampena, Julien Couthouis, Sunjay Sethi, Kathleen McKeever, Michael R Betts, Emil Kakkis, James M Wilson, Samuel Wadsworth, Barbara A Sullivan

{"title":"Prednisolone reduces the interferon response to AAV in cynomolgus macaques and may increase liver gene expression.","authors":"Lili Wang, Claude C Warzecha, Alexander Kistner, Jessica A Chichester, Peter Bell, Elizabeth L Buza, Zhenning He, M Betina Pampena, Julien Couthouis, Sunjay Sethi, Kathleen McKeever, Michael R Betts, Emil Kakkis, James M Wilson, Samuel Wadsworth, Barbara A Sullivan","doi":"10.1016/j.omtm.2022.01.007","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.01.007","url":null,"abstract":"Ornithine transcarbamylase deficiency is a rare X-linked genetic urea cycle disorder leading to episodes of acute hyperammonemia, adverse cognitive and neurological effects, hospitalizations, and in some cases death. DTX301, a non-replicating, recombinant self-complimentary adeno-associated virus vector serotype 8 (scAAV8)-encoding human ornithine transcarbamylase, is a promising gene therapy for ornithine transcarbamylase deficiency; however, the impact of sex and prophylactic immunosuppression on ornithine transcarbamylase gene therapy outcomes is not well characterized. This study sought to describe the impact of sex and immunosuppression in adult, sexually mature female and male cynomolgus macaques through day 140 after DTX301 administration. Four study groups (n = 3/group) were included: male non-immunosuppressed; male immunosuppressed; female non-immunosuppressed; and female immunosuppressed. DTX301 was well tolerated with and without immunosuppression; no notable differences were observed between female and male groups across outcome measures. Prednisolone-treated animals exhibited a trend toward greater vector genome and transgene expression, although the differences were not statistically significant. The hepatic interferon gene signature was significantly decreased in prednisolone-treated animals, and a significant inverse relationship was observed between interferon gene signature levels and hepatic vector DNA and transgene RNA. These observations were not sustained upon immunosuppression withdrawal. Further studies may determine whether the observed effect can be prolonged.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"292-305"},"PeriodicalIF":4.7,"publicationDate":"2022-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/40/fe/main.PMC8841522.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39656247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Rapid, accurate mapping of transgene integration in viable rhesus macaque embryos using enhanced-specificity tagmentation-assisted PCR. 利用增强特异性标记辅助PCR快速、准确地定位活恒河猴胚胎中的转基因整合。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-01-19 eCollection Date: 2022-03-10 DOI: 10.1016/j.omtm.2022.01.009

Junghyun Ryu, William Chan, Jochen M Wettengel, Carol B Hanna, Benjamin J Burwitz, Jon D Hennebold, Benjamin N Bimber

{"title":"Rapid, accurate mapping of transgene integration in viable rhesus macaque embryos using enhanced-specificity tagmentation-assisted PCR.","authors":"Junghyun Ryu, William Chan, Jochen M Wettengel, Carol B Hanna, Benjamin J Burwitz, Jon D Hennebold, Benjamin N Bimber","doi":"10.1016/j.omtm.2022.01.009","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.01.009","url":null,"abstract":"Genome engineering is a powerful tool for in vitro research and the creation of novel model organisms and has growing clinical applications. Randomly integrating vectors, such as lentivirus- or transposase-based methods, are simple and easy to use but carry risks arising from insertional mutagenesis. Here we present enhanced-specificity tagmentation-assisted PCR (esTag-PCR), a rapid and accurate method for mapping transgene integration and copy number. Using stably transfected HepG2 cells, we demonstrate that esTag-PCR has higher integration site detection accuracy and efficiency than alternative tagmentation-based methods. Next, we performed esTag-PCR on rhesus macaque embryos derived from zygotes injected with piggyBac transposase and transposon/transgene plasmid. Using low-input trophectoderm biopsies, we demonstrate that esTag-PCR accurately maps integration events while preserving blastocyst viability. We used these high-resolution data to evaluate the performance of piggyBac-mediated editing of rhesus macaque embryos, demonstrating that increased concentration of transposon/transgene plasmid can increase the fraction of embryos with stable integration; however, the number of integrations per embryo also increases, which may be problematic for some applications. Collectively, esTag-PCR represents an important improvement to the detection of transgene integration, provides a method to validate and screen edited embryos before implantation, and represents an important advance in the creation of transgenic animal models.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"241-254"},"PeriodicalIF":4.7,"publicationDate":"2022-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/7a/f0/main.PMC8829455.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39960379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Delivery of non-viral naked DNA vectors to liver in small weaned pigs by hydrodynamic retrograde intrabiliary injection. 水动力逆行胆道内注射将非病毒裸DNA载体送入小断奶猪肝脏。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-01-19 eCollection Date: 2022-03-10 DOI: 10.1016/j.omtm.2022.01.006

Tatjana Chan, Hiu Man Grisch-Chan, Philipp Schmierer, Ulrike Subotic, Nicole Rimann, Tanja Scherer, Udo Hetzel, Matthias Bozza, Richard Harbottle, James A Williams, Barbara Steblaj, Simone K Ringer, Johannes Häberle, Xaver Sidler, Beat Thöny

{"title":"Delivery of non-viral naked DNA vectors to liver in small weaned pigs by hydrodynamic retrograde intrabiliary injection.","authors":"Tatjana Chan, Hiu Man Grisch-Chan, Philipp Schmierer, Ulrike Subotic, Nicole Rimann, Tanja Scherer, Udo Hetzel, Matthias Bozza, Richard Harbottle, James A Williams, Barbara Steblaj, Simone K Ringer, Johannes Häberle, Xaver Sidler, Beat Thöny","doi":"10.1016/j.omtm.2022.01.006","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.01.006","url":null,"abstract":"Hepatic gene therapy by delivering non-integrating therapeutic vectors in newborns remains challenging due to the risk of dilution and loss of efficacy in the growing liver. Previously we reported on hepatocyte transfection in piglets by intraportal injection of naked DNA vectors. Here, we established delivery of naked DNA vectors to target periportal hepatocytes in weaned pigs by hydrodynamic retrograde intrabiliary injection (HRII). The surgical procedure involved laparotomy and transient isolation of the liver. For vector delivery, a catheter was placed within the common bile duct by enterotomy. Under optimal conditions, no histological abnormalities were observed in liver tissue upon pressurized injections. The transfection of hepatocytes in all tested liver samples was observed with vectors expressing luciferase from a liver-specific promoter. However, vector copy number and luciferase expression were low compared to hydrodynamic intraportal injection. A 10-fold higher number of vector genomes and luciferase expression was observed in pigs using a non-integrating naked DNA vector with the potential for replication. In summary, the HRII application was less efficient (i.e., lower luciferase activity and vector copy numbers) than the intraportal delivery method but was significantly less distressful for the piglets and has the potential for injection (or re-injection) of vector DNA by endoscopic retrograde cholangiopancreatography.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"268-279"},"PeriodicalIF":4.7,"publicationDate":"2022-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/68/e6/main.PMC8829443.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39656246","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Differential T cell immune responses to deamidated adeno-associated virus vector. 不同T细胞对脱酰胺腺相关病毒载体的免疫反应。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-01-18 eCollection Date: 2022-03-10 DOI: 10.1016/j.omtm.2022.01.005

So Jin Bing, Sune Justesen, Wells W Wu, Abdul Mohin Sajib, Stephanee Warrington, Alan Baer, Stephan Thorgrimsen, Rong-Fong Shen, Ronit Mazor

{"title":"Differential T cell immune responses to deamidated adeno-associated virus vector.","authors":"So Jin Bing, Sune Justesen, Wells W Wu, Abdul Mohin Sajib, Stephanee Warrington, Alan Baer, Stephan Thorgrimsen, Rong-Fong Shen, Ronit Mazor","doi":"10.1016/j.omtm.2022.01.005","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.01.005","url":null,"abstract":"Despite the high safety profile demonstrated in clinical trials, the immunogenicity of adeno-associated virus (AAV)-mediated gene therapy remains a major hurdle. Specifically, T-cell-mediated immune responses to AAV vectors are related to loss of efficacy and potential liver toxicities. As post-translational modifications in T cell epitopes have the potential to affect immune reactions, the cellular immune responses to peptides derived from spontaneously deamidated AAV were investigated. Here, we report that highly deamidated sites in AAV9 contain CD4 T cell epitopes with a Th1 cytokine pattern in multiple human donors with diverse human leukocyte antigen (HLA) backgrounds. Furthermore, some peripheral blood mononuclear cell (PBMC) samples demonstrated differential T cell activation to deamidated or non-deamidated epitopes. Also, in vitro and in silico HLA binding assays showed differential binding to the deamidated or non-deamidated peptides in some HLA alleles. This study provides critical attributes to vector-immune-mediated responses, as AAV deamidation can impact the immunogenicity, safety, and efficacy of AAV-mediated gene therapy in some patients.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"255-267"},"PeriodicalIF":4.7,"publicationDate":"2022-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/d7/7a/main.PMC8829777.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39960380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Delivery of nVEGFi using AAV8 for the treatment of neovascular age-related macular degeneration. 使用AAV8递送nVEGFi用于治疗新生血管性年龄相关性黄斑变性。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-01-10 eCollection Date: 2022-03-10 DOI: 10.1016/j.omtm.2022.01.002

Kaiqin She, Jing Su, Qingnan Wang, Yi Liu, Xiaomei Zhong, Xiu Jin, Qinyu Zhao, Jianlu Xiao, Ruiting Li, Hongxin Deng, Fang Lu, Yang Yang, Yuquan Wei

{"title":"Delivery of nVEGFi using AAV8 for the treatment of neovascular age-related macular degeneration.","authors":"Kaiqin She, Jing Su, Qingnan Wang, Yi Liu, Xiaomei Zhong, Xiu Jin, Qinyu Zhao, Jianlu Xiao, Ruiting Li, Hongxin Deng, Fang Lu, Yang Yang, Yuquan Wei","doi":"10.1016/j.omtm.2022.01.002","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.01.002","url":null,"abstract":"Inhibition of vascular endothelial growth factor (VEGF) is the standard therapy for neovascular age-related macular degeneration (nAMD). However, anti-VEGF agents used in the clinic require repeated injections, causing adverse effects. Gene therapy could provide sustained anti-VEGF levels after a single injection, thereby drastically decreasing the treatment burden and improving visual outcomes. In this study, we developed a novel VEGF Trap, nVEGFi, containing domains 1 and 2 of VEGFR1 and domain 3 of VEGFR2 fused to the Fc portion of human IgG. The nVEGFi had a higher expression level than aflibercept under the same expression cassettes of adeno-associated virus (AAV)8 in vitro and in vivo. nVEGFi was found to be noninferior to aflibercept in binding and blocking VEGF in vitro. AAV8-mediated expression of nVEGFi was maintained for at least 12 weeks by subretinal delivery in C57BL/6J mice. In a mouse laser-induced choroidal neovascularization (CNV) model, 4 × 108 genome copies of AAV8-nVEGFi exhibited a significantly increased reduction in the CNV area compared with AAV8-aflibercept (78.1% vs. 63.9%, p < 0.05), while causing no structural or functional changes to the retina. In conclusion, this preclinical study showed that subretinal injection of AAV8-nVEGFi was long lasting, well tolerated, and effective for nAMD treatment, supporting future translation to the clinic.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"210-221"},"PeriodicalIF":4.7,"publicationDate":"2022-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/34/06/main.PMC8800040.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39905118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4