Gene Therapy最新文献_第10页

Sildenafil increases AAV9 transduction after a systemic administration and enhances AAV9-dystrophin therapeutic effect in mdx mice 西地那非可增加全身给药后的 AAV9 转导，并增强 AAV9-dystrophin 对 mdx 小鼠的治疗效果。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-07-27 DOI: 10.1038/s41434-023-00411-3

Kaiyi Zhou, Meng Yuan, Jiabao Sun, Feixu Zhang, Xiaoying Zong, Zhanao Li, Dingyue Tang, Lichen Zhou, Jing Zheng, Xiao Xiao, Xia Wu

{"title":"Sildenafil increases AAV9 transduction after a systemic administration and enhances AAV9-dystrophin therapeutic effect in mdx mice","authors":"Kaiyi Zhou, Meng Yuan, Jiabao Sun, Feixu Zhang, Xiaoying Zong, Zhanao Li, Dingyue Tang, Lichen Zhou, Jing Zheng, Xiao Xiao, Xia Wu","doi":"10.1038/s41434-023-00411-3","DOIUrl":"10.1038/s41434-023-00411-3","url":null,"abstract":"Adeno-associated virus (AAV) vectors have been successfully used to deliver genes for treating rare diseases. However, the systemic administration of high AAV vector doses triggers several adverse effects, including immune response, the asymptomatic elevation of liver transaminase levels, and complement activation. Thus, improving AAV transduction and reducing AAV dosage for treatment is necessary. Recently, we found that a phosphodiesterase-5 inhibitor significantly promoted AAV9 transduction in vitro by regulating the caveolae and macropinocytosis pathways. When AAV9-Gaussian luciferase (AAV9-Gluc) and AAV9-green fluorescent protein (AAV9-GFP) were injected intravenously into mice pre-treated with sildenafil, the expressions of Gluc in the plasma and GFP in muscle tissues significantly increased (P < 0.05). Sildenafil also improved Evans blue permeation in tissues. Additionally, we found that sildenafil promoted Treg proliferation, inhibited B-cell activation, and decreased anti-AAV9 IgG levels (P < 0.05). Furthermore, sildenafil significantly promoted Duchenne muscular dystrophy gene therapy efficacy using AAV9 in mdx mice; it increased micro-dystrophin gene expression, forelimb grip strength, and time spent on the rotarod test, decreased serum creatine kinase levels, and ameliorated histopathology by improving muscle cell morphology and reducing fibrosis (P < 0.05). These results show that sildenafil significantly improved AAV transduction, suppressed the levels of anti-AAV9 IgG, and enhanced the efficacy of gene therapy.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10259421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Correction: Site-specific modifications to AAV8 capsid yields enhanced brain transduction in the neonatal MPS IIIB mouse 更正：对AAV8衣壳的位点特异性修饰增强了新生MPS IIIB小鼠的脑转导。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-07-27 DOI: 10.1038/s41434-023-00412-2

Janine A. Gilkes, Benjamin L. Judkins, Brontie N. Herrera, Ronald J. Mandel, Sanford L. Boye, Shannon E. Boye, Arun Srivastava, Coy D. Heldermon

引用次数: 0

Immunobiology of a rationally-designed AAV2 capsid following intravitreal delivery in mice 小鼠玻璃体内给药后合理设计的AAV2衣壳的免疫生物学。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-06-29 DOI: 10.1038/s41434-023-00409-x

Michael Whitehead, Andrew Sage, Tom Burgoyne, Andrew Osborne, Patrick Yu-Wai-Man, Keith R. Martin

{"title":"Immunobiology of a rationally-designed AAV2 capsid following intravitreal delivery in mice","authors":"Michael Whitehead, Andrew Sage, Tom Burgoyne, Andrew Osborne, Patrick Yu-Wai-Man, Keith R. Martin","doi":"10.1038/s41434-023-00409-x","DOIUrl":"10.1038/s41434-023-00409-x","url":null,"abstract":"Adeno-associated virus serotype 2 (AAV2) is a viral vector that can be used to deliver therapeutic genes to diseased cells in the retina. One strategy for altering AAV2 vectors involves the mutation of phosphodegron residues, which are thought to be phosphorylated/ubiquitinated in the cytosol, facilitating degradation of the vector and the inhibition of transduction. As such, mutation of phosphodegron residues have been correlated with increased transduction of target cells, however, an assessment of the immunobiology of wild-type and phosphodegron mutant AAV2 vectors following intravitreal (IVT) delivery to immunocompetent animals is lacking in the current literature. In this study, we show that IVT of a triple phosphodegron mutant AAV2 capsid is associated with higher levels of humoral immune activation, infiltration of CD4 and CD8 T-cells into the retina, generation of splenic germinal centre reactions, activation of conventional dendritic cell subsets, and elevated retinal gliosis compared to wild-type AAV2 capsids. However, we did not detect significant changes in electroretinography arising after vector administration. We also demonstrate that the triple AAV2 mutant capsid is less susceptible to neutralisation by soluble heparan sulphate and anti-AAV2 neutralising antibodies, highlighting a possible utility for the vector in terms of circumventing pre-existing humoral immunity. In summary, the present study highlights novel aspects of rationally-designed vector immunobiology, which may be relevant to their application in preclinical and clinical settings.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10506909/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10352016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhanced expression of the human Survival motor neuron 1 gene from a codon-optimised cDNA transgene in vitro and in vivo 通过密码子优化的 cDNA 转基因增强人类生存运动神经元 1 基因在体外和体内的表达。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-06-15 DOI: 10.1038/s41434-023-00406-0

Neda A. M. Nafchi, Ellie M. Chilcott, Sharon Brown, Heidi R. Fuller, Melissa Bowerman, Rafael J. Yáñez-Muñoz

{"title":"Enhanced expression of the human Survival motor neuron 1 gene from a codon-optimised cDNA transgene in vitro and in vivo","authors":"Neda A. M. Nafchi, Ellie M. Chilcott, Sharon Brown, Heidi R. Fuller, Melissa Bowerman, Rafael J. Yáñez-Muñoz","doi":"10.1038/s41434-023-00406-0","DOIUrl":"10.1038/s41434-023-00406-0","url":null,"abstract":"Spinal muscular atrophy (SMA) is a neuromuscular disease particularly characterised by degeneration of ventral motor neurons. Survival motor neuron (SMN) 1 gene mutations cause SMA, and gene addition strategies to replace the faulty SMN1 copy are a therapeutic option. We have developed a novel, codon-optimised hSMN1 transgene and produced integration-proficient and integration-deficient lentiviral vectors with cytomegalovirus (CMV), human synapsin (hSYN) or human phosphoglycerate kinase (hPGK) promoters to determine the optimal expression cassette configuration. Integrating, CMV-driven and codon-optimised hSMN1 lentiviral vectors resulted in the highest production of functional SMN protein in vitro. Integration-deficient lentiviral vectors also led to significant expression of the optimised transgene and are expected to be safer than integrating vectors. Lentiviral delivery in culture led to activation of the DNA damage response, in particular elevating levels of phosphorylated ataxia telangiectasia mutated (pATM) and γH2AX, but the optimised hSMN1 transgene showed some protective effects. Neonatal delivery of adeno-associated viral vector (AAV9) vector encoding the optimised transgene to the Smn2B/− mouse model of SMA resulted in a significant increase of SMN protein levels in liver and spinal cord. This work shows the potential of a novel codon-optimised hSMN1 transgene as a therapeutic strategy for SMA.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s41434-023-00406-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9639096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tolerability and tropism of recombinant adeno-associated virus vectors in the African green monkey (Chlorocebus sabaeus) anterior chamber 重组腺相关病毒载体在非洲绿猴（Chlorocebus sabaeus）前房中的耐受性和嗜性。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-05-24 DOI: 10.1038/s41434-023-00407-z

Kristina J. Chern, Kimicia Z. Issac, Zendorf D. Gumbs, Merissa E. O’Connor, Matthew S. Lawrence, Daniel M. Lipinski

{"title":"Tolerability and tropism of recombinant adeno-associated virus vectors in the African green monkey (Chlorocebus sabaeus) anterior chamber","authors":"Kristina J. Chern, Kimicia Z. Issac, Zendorf D. Gumbs, Merissa E. O’Connor, Matthew S. Lawrence, Daniel M. Lipinski","doi":"10.1038/s41434-023-00407-z","DOIUrl":"10.1038/s41434-023-00407-z","url":null,"abstract":"While many studies have investigated the use of recombinant adeno-associated vectors (rAAV) in the posterior chamber for treatment of inherited retinal diseases, fewer studies have looked at rAAV’s ability to transduce cells within the anterior chamber. This study focuses on evaluating the tropism and tolerability of three rAAV serotypes—rAAV2/6, rAAV2/9, and rAAV2/2[MAX]—expressing a green fluorescent protein (GFP) reporter following intracameral injection in the non-human primate (NHP) African green monkey (Chlorocebus sabaeus) model. Injection of high dose (1 × 1012 vg/eye) rAAV vector resulted in transient inflammation characterized by aqueous flare and cellular infiltrate that resolved without intervention in all serotypes. Post-mortem histology revealed widespread expression of GFP in cells of the trabecular meshwork and iris in high dose rAAV2/6, rAAV2/9, and particularly rAAV2/2[MAX] eyes, indicating that rAAV vectors of these serotypes have broad tropism for cells of the anterior chamber and may facilitate the treatment of blinding disorders, such as glaucoma.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10663143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

RapaCaspase-9-based suicide gene applied to the safety of IL-1RAP CAR-T cells 基于RapaCaspase-9的自杀基因应用于IL-1RAP CAR-T细胞的安全性。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-05-12 DOI: 10.1038/s41434-023-00404-2

Lucie Bouquet, Elodie Bôle-Richard, Walid Warda, Mathieu Neto Da Rocha, Rim Trad, Clémentine Nicod, Rafik Haderbache, Delphine Genin, Christophe Ferrand, Marina Deschamps

{"title":"RapaCaspase-9-based suicide gene applied to the safety of IL-1RAP CAR-T cells","authors":"Lucie Bouquet, Elodie Bôle-Richard, Walid Warda, Mathieu Neto Da Rocha, Rim Trad, Clémentine Nicod, Rafik Haderbache, Delphine Genin, Christophe Ferrand, Marina Deschamps","doi":"10.1038/s41434-023-00404-2","DOIUrl":"10.1038/s41434-023-00404-2","url":null,"abstract":"Even if adoptive cell transfer (ACT) has already shown great clinical efficiency in different types of disease, such as cancer, some adverse events consistently occur, and suicide genes are an interesting system to manage these events. Our team developed a new medical drug candidate, a chimeric antigen receptor (CAR) targeting interleukin-1 receptor accessory protein (IL-1RAP), which needs to be evaluated in clinical trials with a clinically applicable suicide gene system. To prevent side effects and ensure the safety of our candidate, we devised two constructs carrying an inducible suicide gene, RapaCasp9-G or RapaCasp9-A, containing a single-nucleotide polymorphism (rs1052576) affecting the efficiency of endogenous caspase 9. These suicide genes are activated by rapamycin and based on the fusion of human caspase 9 with a modified human FK-binding protein, allowing conditional dimerization. RapaCasp9-G- and RapaCasp9-A-expressing gene-modified T cells (GMTCs) were produced from healthy donors (HDs) and acute myeloid leukemia (AML) donors. The RapaCasp9-G suicide gene demonstrated better efficiency, and we showed its in vitro functionality in different clinically relevant culture conditions. Moreover, as rapamycin is not pharmacologically inert, we also demonstrated its safe use as part of our therapy.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10506905/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10297871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Repeat or single-dose lentiviral vector administration to mouse lungs? It’s all about the timing 小鼠肺部重复或单剂量慢病毒载体给药？这一切都与时间有关。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-05-10 DOI: 10.1038/s41434-023-00403-3

Martin Donnelley, Patricia Cmielewski, Emma Knight, Chantelle Carpentieri, Alexandra McCarron, Nathan Rout-Pitt, David Parsons, Nigel Farrow

{"title":"Repeat or single-dose lentiviral vector administration to mouse lungs? It’s all about the timing","authors":"Martin Donnelley, Patricia Cmielewski, Emma Knight, Chantelle Carpentieri, Alexandra McCarron, Nathan Rout-Pitt, David Parsons, Nigel Farrow","doi":"10.1038/s41434-023-00403-3","DOIUrl":"10.1038/s41434-023-00403-3","url":null,"abstract":"Lentiviral vectors are attractive delivery vehicles for cystic fibrosis gene therapy owing to their low immunogenicity and ability to integrate into the host cell genome, thereby producing long-term, stable gene expression. Nonetheless, repeat dosing may be required to increase initial expression levels, and/or boost levels when they wane. The primary aim of this study was to determine if repeat dosing of a VSV-G pseudotyped LV vector delivered into mouse lungs is more effective than a single dose. C57Bl/6 mouse lungs were conditioned with lysophosphatidylcholine, followed one-hour later by a LV vector carrying the luciferase reporter gene, using six different short-term (≤1 wk) and long-term (>1 wk) dosing schedules. Luciferase expression was quantified using bioluminescence imaging over 12 months. Most dosing schedules produced detectable bioluminescence over the 12-month period, but the shorter intervals (≤1 wk) produced higher levels of flux than the longest interval (five doses at least 1-month apart). Ex vivo lung analysis at 12 months showed that the estimated mean flux for the group that received two doses 1-week apart was significantly greater than the single dose group and the two groups that received doses over a period greater than 1-week. These results suggest that early consecutive multiple doses are more effective at improving gene expression in mouse lungs at 12 months, than longer repeat dosing intervals.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10506910/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10349807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Adeno-associated virus vectors and neurotoxicity-lessons from preclinical and human studies. 腺相关病毒载体和神经毒性的临床前和人体研究经验教训。

IF 4.6 3区医学

Gene Therapy Pub Date : 2023-05-10 DOI: 10.1038/s41434-023-00405-1

Daniel Stone, Martine Aubert, Keith R Jerome

{"title":"Adeno-associated virus vectors and neurotoxicity-lessons from preclinical and human studies.","authors":"Daniel Stone, Martine Aubert, Keith R Jerome","doi":"10.1038/s41434-023-00405-1","DOIUrl":"10.1038/s41434-023-00405-1","url":null,"abstract":"<p><p>Over 15 years after hepatotoxicity was first observed following administration of an adeno-associated virus (AAV) vector during a hemophilia B clinical trial, recent reports of treatment-associated neurotoxicity in animals and humans have brought the potential impact of AAV-associated toxicity back to prominence. In both pre-clinical studies and clinical trials, systemic AAV administration has been associated with neurotoxicity in peripheral nerve ganglia and spinal cord. Neurological signs have also been seen following direct AAV injection into the brain, both in non-human primates and in a clinical trial for late infantile Batten disease. Neurotoxic events appear variable across species, and preclinical animal studies do not fully predict clinical observations. Accumulating data suggest that AAV-associated neurotoxicity may be underdiagnosed and may differ between species in terms of frequency and/or severity. In this review, we discuss the different animal models that have been used to demonstrate AAV-associated neurotoxicity, its potential causes and consequences, and potential approaches to blunt AAV-associated neurotoxicity.</p>","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":4.6,"publicationDate":"2023-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11247785/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9524917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Review of cardiac safety in onasemnogene abeparvovec gene replacement therapy: translation from preclinical to clinical findings onaseminogene abeparvovec基因替代疗法的心脏安全性综述：从临床前到临床结果的转化。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-04-25 DOI: 10.1038/s41434-023-00401-5

Deepa H. Chand, Rui Sun, Karim A. Diab, Damien Kenny, Francis Fonyuy Tukov

{"title":"Review of cardiac safety in onasemnogene abeparvovec gene replacement therapy: translation from preclinical to clinical findings","authors":"Deepa H. Chand, Rui Sun, Karim A. Diab, Damien Kenny, Francis Fonyuy Tukov","doi":"10.1038/s41434-023-00401-5","DOIUrl":"10.1038/s41434-023-00401-5","url":null,"abstract":"Human gene replacement therapies such as onasemnogene abeparvovec (OA) use recombinant adeno-associated virus (rAAV) vectors to treat monogenic disorders. The heart and liver are known target organs of toxicity in animals; with cardiac and hepatic monitoring recommended in humans after OA dosing. This manuscript provides a comprehensive description of cardiac data from preclinical studies and clinical sources including clinical trials, managed access programs and the post-marketing setting following intravenous OA administration through 23 May 2022. Single dose mouse GLP-Toxicology studies revealed dose-dependent cardiac findings including thrombi, myocardial inflammation and degeneration/regeneration, which were associated with early mortality (4-7 weeks) in the high dose groups. No such findings were documented in non-human primates (NHP) after 6 weeks or 6 months post-dose. No electrocardiogram or echocardiogram abnormalities were noted in NHP or humans. After OA dosing, some patients developed isolated elevations in troponin without associated signs/symptoms; the reported cardiac adverse events in patients were considered of secondary etiology (e.g. respiratory dysfunction or sepsis leading to cardiac events). Clinical data indicate cardiac toxicity observed in mice does not translate to humans. Cardiac abnormalities have been associated with SMA. Healthcare professionals should use medical judgment when evaluating the etiology and assessment of cardiac events post OA dosing so as to consider all possibilities and manage the patient accordingly.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10125853/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10299141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

rAAV-PHP.B escapes the mouse eye and causes lethality whereas rAAV9 can transduce aniridic corneal limbal stem cells without lethality rAAV-PHP.B逃离小鼠眼睛并引起致命性，而rAAV9可以转导无虹膜角膜缘干细胞而没有致命性。

IF 5.1 3区医学

Gene Therapy Pub Date : 2023-04-19 DOI: 10.1038/s41434-023-00400-6

Seyedeh Zeinab Mirjalili Mohanna, Andrea J. Korecki, Elizabeth M. Simpson

{"title":"rAAV-PHP.B escapes the mouse eye and causes lethality whereas rAAV9 can transduce aniridic corneal limbal stem cells without lethality","authors":"Seyedeh Zeinab Mirjalili Mohanna, Andrea J. Korecki, Elizabeth M. Simpson","doi":"10.1038/s41434-023-00400-6","DOIUrl":"10.1038/s41434-023-00400-6","url":null,"abstract":"Recently safety concerns have been raised in connection with high doses of recombinant adeno-associated viruses (rAAV). Therefore, we undertook a series of experiments to test viral capsid (rAAV9 and rAAV-PHP.B), dose, and route of administration (intrastromal, intravitreal, and intravenous) focused on aniridia, a congenital blindness that currently has no cure. The success of gene therapy for aniridia may depend on the presence of functional limbal stem cells (LSCs) in the damaged aniridic corneas and whether rAAV can transduce them. Both these concerns were unknown, and thus were also addressed by our studies. For the first time, we report ataxia and lethality after intravitreal or intrastromal rAAV-PHP.B virus injections. We demonstrated virus escape from the eye and transduction of non-ocular tissues by rAAV9 and rAAV-PHP.B capsids. We have also shown that intrastromal and intravitreal delivery of rAAV9 can transduce functional LSCs, as well as all four PAX6-expressing retinal cell types in aniridic eye, respectively. Overall, lack of adverse events and successful transduction of LSCs and retinal cells makes it clear that rAAV9 is the capsid of choice for future aniridia gene therapy. Our finding of rAAV lethality after intraocular injections will be impactful for other researchers developing rAAV-based gene therapies.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":null,"pages":null},"PeriodicalIF":5.1,"publicationDate":"2023-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10506911/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10661836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0