Molecular Therapy. Methods & Clinical Development最新文献_第4页

Therapeutic efficacy of rscAAVrh74.miniCMV.LIPA gene therapy in a mouse model of lysosomal acid lipase deficiency. rscAAVrh74.miniCMV的治疗效果。溶酶体酸性脂肪酶缺乏症小鼠模型的LIPA基因治疗。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-08-04 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.08.001

Patricia Lam, Anna Ashbrook, Deborah A Zygmunt, Cong Yan, Hong Du, Paul T Martin

{"title":"Therapeutic efficacy of rscAAVrh74.miniCMV.LIPA gene therapy in a mouse model of lysosomal acid lipase deficiency.","authors":"Patricia Lam, Anna Ashbrook, Deborah A Zygmunt, Cong Yan, Hong Du, Paul T Martin","doi":"10.1016/j.omtm.2022.08.001","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.08.001","url":null,"abstract":"Lysosomal acid lipase deficiency (LAL-D) presents as one of two rare autosomal recessive diseases: Wolman disease (WD), a severe disorder presenting in infancy characterized by absent or very low LAL activity, and cholesteryl ester storage disease (CESD), a less severe, later onset disease form. Recent clinical studies have shown efficacy of enzyme replacement therapy for both forms of LAL-D; however, no gene therapy approach has yet been developed for clinical use. Here, we show that rscAAVrh74.miniCMV.LIPA gene therapy can significantly improve disease symptoms in the Lipa -/- mouse model of LAL-D. Treatment dramatically lowered hepatosplenomegaly, liver and spleen triglyceride and cholesterol levels, and serum expression of markers of liver damage. Measures of liver inflammation and fibrosis were also reduced. Treatment of young adult mice was more effective than treatment of neonates, and enzyme activity was elevated in serum, consistent with possible bystander effects. These results demonstrate that adeno associated virus (AAV)-mediated LIPA gene-replacement therapy may be a viable option to treat patients with LAL-D, particularly patients with CESD.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"413-426"},"PeriodicalIF":4.7,"publicationDate":"2022-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9403906/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33467129","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

Immunogenicity assessment of AAV-based gene therapies: An IQ consortium industry white paper. 基于aav的基因疗法的免疫原性评估:IQ联盟行业白皮书。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-08-02 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.018

Tong-Yuan Yang, Manuela Braun, Wibke Lembke, Fraser McBlane, John Kamerud, Stephen DeWall, Edit Tarcsa, Xiaodong Fang, Lena Hofer, Uma Kavita, Vijay V Upreti, Swati Gupta, LiNa Loo, Alison J Johnson, Rakesh Kantilal Chandode, Kay-Gunnar Stubenrauch, Maya Vinzing, Cindy Q Xia, Vibha Jawa

{"title":"Immunogenicity assessment of AAV-based gene therapies: An IQ consortium industry white paper.","authors":"Tong-Yuan Yang, Manuela Braun, Wibke Lembke, Fraser McBlane, John Kamerud, Stephen DeWall, Edit Tarcsa, Xiaodong Fang, Lena Hofer, Uma Kavita, Vijay V Upreti, Swati Gupta, LiNa Loo, Alison J Johnson, Rakesh Kantilal Chandode, Kay-Gunnar Stubenrauch, Maya Vinzing, Cindy Q Xia, Vibha Jawa","doi":"10.1016/j.omtm.2022.07.018","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.018","url":null,"abstract":"Immunogenicity has imposed a challenge to efficacy and safety evaluation of adeno-associated virus (AAV) vector-based gene therapies. Mild to severe adverse events observed in clinical development have been implicated with host immune responses against AAV gene therapies, resulting in comprehensive evaluation of immunogenicity during nonclinical and clinical studies mandated by health authorities. Immunogenicity of AAV gene therapies is complex due to the number of risk factors associated with product components and pre-existing immunity in human subjects. Different clinical mitigation strategies have been employed to alleviate treatment-induced or -boosted immunogenicity in order to achieve desired efficacy, reduce toxicity, or treat more patients who are seropositive to AAV vectors. In this review, the immunogenicity risk assessment, manifestation of immunogenicity and its impact in nonclinical and clinical studies, and various clinical mitigation strategies are summarized. Last, we present bioanalytical strategies, methodologies, and assay validation applied to appropriately monitor immunogenicity in AAV gene therapy-treated subjects.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"471-494"},"PeriodicalIF":4.7,"publicationDate":"2022-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/dd/20/main.PMC9418752.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33461175","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: 11

Preclinical safety and efficacy of lentiviral-mediated gene therapy for leukocyte adhesion deficiency type I. 慢病毒介导的基因治疗I型白细胞粘附缺陷的临床前安全性和有效性。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-08-01 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.015

Cristina Mesa-Núñez, Carlos Damián, María Fernández-García, Begoña Díez, Gayatri Rao, Jonathan D Schwartz, Ken M Law, Julián Sevilla, Paula Río, Rosa Yáñez, Juan A Bueren, Elena Almarza

{"title":"Preclinical safety and efficacy of lentiviral-mediated gene therapy for leukocyte adhesion deficiency type I.","authors":"Cristina Mesa-Núñez, Carlos Damián, María Fernández-García, Begoña Díez, Gayatri Rao, Jonathan D Schwartz, Ken M Law, Julián Sevilla, Paula Río, Rosa Yáñez, Juan A Bueren, Elena Almarza","doi":"10.1016/j.omtm.2022.07.015","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.015","url":null,"abstract":"Leukocyte adhesion deficiency type I (LAD-I) is a primary immunodeficiency caused by mutations in the ITGB2 gene, which encodes for the CD18 subunit of β2-integrins. Deficient expression of β2-integrins results in impaired neutrophil migration in response to bacterial and fungal infections. Using a lentiviral vector (LV) that mediates a preferential myeloid expression of human CD18 (Chim.hCD18-LV), we first demonstrated that gene therapy efficiently corrected the phenotype of mice with severe LAD-I. Next, we investigated if the ectopic hCD18 expression modified the phenotypic characteristics of human healthy donor hematopoietic stem cells and their progeny. Significantly, transduction of healthy CD34+ cells with the Chim.hCD18-LV did not modify the membrane expression of CD18 nor the adhesion of physiological ligands to transduced cells. Additionally, we observed that the repopulating properties of healthy CD34+ cells were preserved following transduction with the Chim.hCD18-LV, and that a safe polyclonal repopulation pattern was observed in transplanted immunodeficient NOD scid gamma (NSG) mice. In a final set of experiments, we demonstrated that transduction of CD34+ cells from a severe LAD-I patient with the Chim.hCD18-LV restores the expression of β2-integrins in these cells. These results offer additional preclinical safety and efficacy evidence supporting the gene therapy of patients with severe LAD-I.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"459-470"},"PeriodicalIF":4.7,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/64/32/main.PMC9418989.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33460696","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

Targeting the lung epithelium after intravenous delivery by directed evolution of underexplored sites on the AAV capsid. 通过AAV衣壳上未开发部位的定向进化靶向静脉给药后的肺上皮。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-08-01 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.010

David Goertsen, Nick Goeden, Nicholas C Flytzanis, Viviana Gradinaru

引用次数: 7

Efficient production of inhibitor-free foamy virus glycoprotein-containing retroviral vectors by proteoglycan-deficient packaging cells. 缺乏蛋白聚糖的包装细胞高效生产无抑制剂泡沫病毒糖蛋白逆转录病毒载体。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-08-01 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.004

Clara Marie Munz, Henriette Kreher, Alexander Erdbeer, Stefanie Richter, Dana Westphal, Buqing Yi, Rayk Behrendt, Nicole Stanke, Fabian Lindel, Dirk Lindemann

{"title":"Efficient production of inhibitor-free foamy virus glycoprotein-containing retroviral vectors by proteoglycan-deficient packaging cells.","authors":"Clara Marie Munz, Henriette Kreher, Alexander Erdbeer, Stefanie Richter, Dana Westphal, Buqing Yi, Rayk Behrendt, Nicole Stanke, Fabian Lindel, Dirk Lindemann","doi":"10.1016/j.omtm.2022.07.004","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.004","url":null,"abstract":"Foamy viruses (FVs) or heterologous retroviruses pseudotyped with FV glycoprotein enable transduction of a great variety of target tissues of disparate species. Specific cellular entry receptors responsible for this exceptionally broad tropism await their identification. Though, ubiquitously expressed heparan sulfate proteoglycan (HS-PG) is known to serve as an attachment factor of FV envelope (Env)-containing virus particles, greatly enhancing target cell permissiveness. Production of high-titer, FV Env-containing retroviral vectors is strongly dependent on the use of cationic polymer-based transfection reagents like polyethyleneimine (PEI). We identified packaging cell-surface HS-PG expression to be responsible for this requirement. Efficient release of FV Env-containing virus particles necessitates neutralization of HS-PG binding sites by PEI. Remarkably, remnants of PEI in FV Env-containing vector supernatants, which are not easily removable, negatively impact target cell transduction, in particular those of myeloid and lymphoid origin. To overcome this limitation for production of FV Env-containing retrovirus supernatants, we generated 293T-based packaging cell lines devoid of HS-PG by genome engineering. This enabled, for the first, time production of inhibitor-free, high-titer FV Env-containing virus supernatants by non-cationic polymer-mediated transfection. Depending on the type of virus, produced titers were 2- to 10-fold higher compared with those obtained by PEI transfection.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"394-412"},"PeriodicalIF":4.7,"publicationDate":"2022-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ab/77/main.PMC9388887.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33444210","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: 0

Structural basis of receptor usage by the engineered capsid AAV-PHP.eB. 工程衣壳AAV-PHP.eB受体使用的结构基础。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-07-31 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.011

Seongmin Jang, Hao K Shen, Xiaozhe Ding, Timothy F Miles, Viviana Gradinaru

{"title":"Structural basis of receptor usage by the engineered capsid AAV-PHP.eB.","authors":"Seongmin Jang, Hao K Shen, Xiaozhe Ding, Timothy F Miles, Viviana Gradinaru","doi":"10.1016/j.omtm.2022.07.011","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.011","url":null,"abstract":"Adeno-associated virus serotype 9 (AAV9) is a promising gene therapy vector for treating neurodegenerative diseases due to its ability to penetrate the blood-brain barrier. PHP.eB was engineered from AAV9 by insertion of a 7-amino acid peptide and point mutation of neighboring residues, thereby enhancing potency in the central nervous system. Here, we report a 2.24-Å resolution cryo-electron microscopy structure of PHP.eB, revealing conformational differences from other 7-mer insertion capsid variants. In PHP.eB, the 7-mer loop adopts a bent conformation, mediated by an interaction between engineered lysine and aspartate residues. Further, we identify PKD2 as the main AAV receptor (AAVR) domain recognizing both AAV9 and PHP.eB and find that the PHP.eB 7-mer partially destabilizes this interaction. Analysis of previously reported AAV structures together with our pull-down data demonstrate that the 7-mer topology determined by the lysine-aspartate interaction dictates AAVR binding strength. Our results suggest that PHP.eB's altered tropism may arise from both an additional interaction with LY6A and weakening of its AAVR interaction. Changing the insertion length, but not sequence, modifies PKD2 binding affinity, suggesting that a steric clash impedes AAVR binding. This research suggests improved library designs for future AAV selections to identify non-LY6A-dependent vectors and modulate AAVR interaction strength.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"343-354"},"PeriodicalIF":4.7,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/22/e8/main.PMC9382559.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33444208","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: 7

Pre-existing anti-drug antibodies in Fabry disease show less affinity for pegunigalsidase alfa. 法布里病中预先存在的抗药物抗体对pegunigalsidase alfa的亲和力较低。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-07-31 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.009

Malte Lenders, Solvey Pollmann, Melina Terlinden, Eva Brand

{"title":"Pre-existing anti-drug antibodies in Fabry disease show less affinity for pegunigalsidase alfa.","authors":"Malte Lenders, Solvey Pollmann, Melina Terlinden, Eva Brand","doi":"10.1016/j.omtm.2022.07.009","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.009","url":null,"abstract":"We analyzed the cross-reactivity of anti-drug antibodies (ADAs) against agalsidase-alfa and -beta from 49 patients with Fabry disease (FD) against the novel PEGylated enzyme pegunigalsidase-alfa (PRX-102). The affinity of purified anti-AGAL antibodies from pooled patient sera was significantly lower for PRX-102 compared to agalsidase-alfa and -beta (both p < 0.05). Pull-down experiments revealed the presence of masked epitopes on PRX-102, possibly due to PEGylation. ADA titers in serum (μg/mL) and corresponding inhibitory capacities against agalsidase-alfa and -beta were measured in male patients with FD, showing strong correlations (r2 = 0.9978 and 0.4930, both p < 0.001). Affinities of ADAs of individual patients against PRX-102 (Kd: 3.55 ± 2.72 μmol) were significantly lower compared to agalsidase alfa (Kd: 1.99 ± 1.26 μmol) and -beta (Kd: 2.18 ± 1.51 μmol) (both p < 0.0001). Cross-ELISAs supported the presence of masked epitopes on PRX-102. Importantly, inhibition measurements also revealed a 30% reduction in inhibitory capacity of pre-existing ADAs towards PRX-102. Enzyme-uptake experiments in AGAL-deficient EA.hy926 cells demonstrated less effects of ADAs on cellular PRX-102 uptake compared with agalsidase beta. We conclude that due to the reduced affinity of pre-existing ADAs against agalsidase-alfa or -beta, ADA-affected patients might benefit from a therapy switch to PRX-102, which is currently evaluated in clinical trials.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"323-330"},"PeriodicalIF":4.7,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/04/b6/main.PMC9379515.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40628721","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: 8

Single-dose AAV vector gene immunotherapy to treat food allergy. 单剂量AAV载体基因免疫疗法治疗食物过敏。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-07-16 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.008

Miguel Gonzalez-Visiedo, Xin Li, Maite Munoz-Melero, Michael D Kulis, Henry Daniell, David M Markusic

{"title":"Single-dose AAV vector gene immunotherapy to treat food allergy.","authors":"Miguel Gonzalez-Visiedo, Xin Li, Maite Munoz-Melero, Michael D Kulis, Henry Daniell, David M Markusic","doi":"10.1016/j.omtm.2022.07.008","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.008","url":null,"abstract":"Immunotherapies for patients with food allergy have shown some success in limiting allergic responses. However, these approaches require lengthy protocols with repeated allergen dosing and patients can relapse following discontinuation of treatment. The purpose of this study was to test if a single dose of an adeno-associated virus (AAV) vector can safely prevent and treat egg allergy in a mouse model. AAV vectors expressing ovalbumin (OVA) under an ubiquitous or liver-specific promoter were injected prior to or after epicutaneous sensitization with OVA. Mice treated with either AAV8-OVA vector were completely protected from allergy sensitization. These animals had a significant reduction in anaphylaxis mediated by a reduction in OVA-specific IgE titers. In mice with established OVA allergy, allergic responses were mitigated only in mice treated with an AAV8-OVA vector expressing OVA from an ubiquitous promoter. In conclusion, an AAV vector with a liver-specific promoter was more effective for allergy prevention, but higher OVA levels were necessary for reducing symptoms in preexisting allergy. Overall, our AAV gene immunotherapy resulted in an expansion of OVA-specific FoxP3+ CD4+ T cells, an increase in the regulatory cytokine IL-10, and a reduction in the IgE promoting cytokine IL-13.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"309-322"},"PeriodicalIF":4.7,"publicationDate":"2022-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/7d/8e/main.PMC9361215.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40628722","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: 0

mRNA-based therapy proves superior to the standard of care for treating hereditary tyrosinemia 1 in a mouse model. 在小鼠模型中，基于mrna的治疗证明优于治疗遗传性酪氨酸血症1的标准护理。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-07-15 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.006

Maximiliano L Cacicedo, Christine Weinl-Tenbruck, Daniel Frank, Sebastian Wirsching, Beate K Straub, Jana Hauke, Jürgen G Okun, Nigel Horscroft, Julia B Hennermann, Fred Zepp, Frédéric Chevessier-Tünnesen, Stephan Gehring

{"title":"mRNA-based therapy proves superior to the standard of care for treating hereditary tyrosinemia 1 in a mouse model.","authors":"Maximiliano L Cacicedo, Christine Weinl-Tenbruck, Daniel Frank, Sebastian Wirsching, Beate K Straub, Jana Hauke, Jürgen G Okun, Nigel Horscroft, Julia B Hennermann, Fred Zepp, Frédéric Chevessier-Tünnesen, Stephan Gehring","doi":"10.1016/j.omtm.2022.07.006","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.006","url":null,"abstract":"Hereditary tyrosinemia type 1 is an inborn error of amino acid metabolism characterized by deficiency of fumarylacetoacetate hydrolase (FAH). Only limited treatment options (e.g., oral nitisinone) are available. Patients must adhere to a strict diet and face a life-long risk of complications, including liver cancer and progressive neurocognitive decline. There is a tremendous need for innovative therapies that standardize metabolite levels and promise normal development. Here, we describe an mRNA-based therapeutic approach that rescues Fah-deficient mice, a well-established tyrosinemia model. Repeated intravenous or intramuscular administration of lipid nanoparticle-formulated human FAH mRNA resulted in FAH protein synthesis in deficient mouse livers, stabilized body weight, normalized pathologic increases in metabolites after nitisinone withdrawal, and prevented early death. Dose reduction and extended injection intervals proved therapeutically effective. These results provide proof of concept for an mRNA-based therapeutic approach to treating hereditary tyrosinemia type 1 that is superior to the standard of care.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"294-308"},"PeriodicalIF":4.7,"publicationDate":"2022-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/4c/12/main.PMC9357842.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40616547","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

Systemic delivery of an AAV9 exon-skipping vector significantly improves or prevents features of Duchenne muscular dystrophy in the Dup2 mouse. 系统递送AAV9外显子跳跃载体可显著改善或防止Dup2小鼠杜氏肌营养不良的特征。

IF 4.7

Molecular Therapy. Methods & Clinical Development Pub Date : 2022-07-11 eCollection Date: 2022-09-08 DOI: 10.1016/j.omtm.2022.07.005

Nicolas Wein, Tatyana A Vetter, Adeline Vulin, Tabatha R Simmons, Emma C Frair, Adrienne J Bradley, Liubov V Gushchina, Camila F Almeida, Nianyuan Huang, Daniel Lesman, Dhanarajan Rajakumar, Robert B Weiss, Kevin M Flanigan

{"title":"Systemic delivery of an AAV9 exon-skipping vector significantly improves or prevents features of Duchenne muscular dystrophy in the Dup2 mouse.","authors":"Nicolas Wein, Tatyana A Vetter, Adeline Vulin, Tabatha R Simmons, Emma C Frair, Adrienne J Bradley, Liubov V Gushchina, Camila F Almeida, Nianyuan Huang, Daniel Lesman, Dhanarajan Rajakumar, Robert B Weiss, Kevin M Flanigan","doi":"10.1016/j.omtm.2022.07.005","DOIUrl":"https://doi.org/10.1016/j.omtm.2022.07.005","url":null,"abstract":"Duchenne muscular dystrophy (DMD) is typically caused by mutations that disrupt the DMD reading frame, but nonsense mutations in the 5' part of the gene induce utilization of an internal ribosomal entry site (IRES) in exon 5, driving expression of a highly functional N-truncated dystrophin. We have developed an AAV9 vector expressing U7 small nuclear RNAs targeting DMD exon 2 and have tested it in a mouse containing a duplication of exon 2, in which skipping of both exon 2 copies induces IRES-driven expression, and skipping of one copy leads to wild-type dystrophin expression. One-time intravascular injection either at postnatal days 0-1 or at 2 months results in efficient exon skipping and dystrophin expression, and significant protection from functional and pathologic deficits. Immunofluorescence quantification showed 33%-53% average dystrophin intensity and 55%-79% average dystrophin-positive fibers in mice treated in adulthood, with partial amelioration of DMD pathology and correction of DMD-associated alterations in gene expression. In mice treated neonatally, dystrophin immunofluorescence reached 49%-85% of normal intensity and 76%-99% dystrophin-positive fibers, with near-complete correction of dystrophic pathology, and these beneficial effects persisted for at least 6 months. Our results demonstrate the robustness, durability, and safety of exon 2 skipping using scAAV9.U7snRNA.ACCA, supporting its clinical use.","PeriodicalId":517056,"journal":{"name":"Molecular Therapy. Methods & Clinical Development","volume":" ","pages":"279-293"},"PeriodicalIF":4.7,"publicationDate":"2022-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/5c/6b/main.PMC9356240.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40696974","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: 13