{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T17:31:41Z","timestamp":1790098301066,"version":"4.0.1"},"reference-count":29,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,3,5]],"date-time":"2024-03-05T00:00:00Z","timestamp":1709596800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The efficacy of adeno-associated virus (AAV)-based gene therapy is dependent on effective viral transduction, which might be inhibited by preexisting immunity to AAV acquired from infection or maternal delivery. Anti-AAV neutralizing Abs (NAbs) titer is usually measured by in vitro assay and used for patient enroll; however, this assay could not evaluate NAbs\u2019 impacts on AAV pharmacology and potential harm in vivo. Here, we infused a mouse anti-AAV9 monoclonal antibody into Balb\/C mice 2 h before receiving 1.2 \u00d7 1014 or 3 \u00d7 1013 vg\/kg of rAAV9-coGAA by tail vein, a drug for our ongoing clinical trials for Pompe disease. The pharmacokinetics, pharmacodynamics, and cellular responses combined with in vitro NAb assay validated the different impacts of preexisting NAbs at different levels in vivo. Sustained GAA expression in the heart, liver, diaphragm, and quadriceps were observed. The presence of high-level NAb, a titer about 1:1000, accelerated vector clearance in blood and completely blocked transduction. The AAV-specific T cell responses tended to increase when the titer of NAb exceeded 1:200. A low-level NAbs, near 1:100, had no effect on transduction in the heart and liver as well as cellular responses, but decreased transduction in muscles slightly. Therefore, we propose to preclude patients with NAb titers &gt; 1:100 from rAAV9-coGAA clinical trials.<\/jats:p>","DOI":"10.3390\/v16030400","type":"journal-article","created":{"date-parts":[[2024,3,5]],"date-time":"2024-03-05T03:03:20Z","timestamp":1709607800000},"page":"400","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Using an In Vivo Mouse Model to Determine the Exclusion Criteria of Preexisting Anti-AAV9 Neutralizing Antibody Titer of Pompe Disease Patients in Clinical Trials"],"prefix":"10.3390","volume":"16","author":[{"given":"Hanqing","family":"Wang","sequence":"first","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cengceng","family":"Zhang","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheyue","family":"Dong","sequence":"additional","affiliation":[{"name":"Beijing FivePlus Gene Technology Co., Ltd., Beijing 102629, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xueyang","family":"Zhu","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuchu","family":"Zheng","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyang","family":"Liu","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianfang","family":"Zhou","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuangqing","family":"Yu","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaobing","family":"Wu","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Dong","sequence":"additional","affiliation":[{"name":"Genecradle Therapeutics Inc., Beijing 100176, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1389","DOI":"10.1089\/hum.2011.031","article-title":"Impact of preexisting immunity on gene transfer to nonhuman primate liver with adeno-associated virus 8 vectors","volume":"22","author":"Wang","year":"2011","journal-title":"Hum. Gene Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1038\/mt.2009.245","article-title":"The pleiotropic effects of natural AAV infections on liver-directed gene transfer in macaques","volume":"18","author":"Wang","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3241","DOI":"10.1182\/bloodadvances.2019000811","article-title":"Etranacogene dezaparvovec (AMT-061 phase 2b): Normal\/near normal FIX activity and bleed cessation in hemophilia B","volume":"3","author":"Giermasz","year":"2019","journal-title":"Blood Adv."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1089\/hum.2019.143","article-title":"Prevalence and Relevance of Preexisting Anti-Adeno-Associated Virus Immunity in the Context of Gene Therapy for Crigler-Najjar Syndrome","volume":"30","author":"Aronson","year":"2019","journal-title":"Hum. Gene Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.omtm.2018.02.003","article-title":"Influence of Preexisting Anti-capsid Neutralizing and Binding Antibodies on AAV Vector Transduction","volume":"9","author":"Fitzpatrick","year":"2018","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1038\/mt.2008.76","article-title":"Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection","volume":"16","author":"Zincarelli","year":"2008","journal-title":"Mol. Ther."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1007\/s13311-018-0655-y","article-title":"Pompe Disease: From Basic Science to Therapy","volume":"15","author":"Kohler","year":"2018","journal-title":"Neurotherapeutics"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Meena, N.K., and Raben, N. (2020). Pompe Disease: New Developments in an Old Lysosomal Storage Disorder. Biomolecules, 10.","DOI":"10.3390\/biom10091339"},{"key":"ref_9","first-page":"24","article-title":"AAV Vector Mediated Gene Therapy in Pompe Model Mice","volume":"42","author":"Wu","year":"2022","journal-title":"China Biotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.omtm.2019.11.009","article-title":"TPP Combined with DGUC as an Economic and Universal Process for Large-Scale Purification of AAV Vectors","volume":"17","author":"Yu","year":"2020","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_11","first-page":"132","article-title":"Biochemical, immunological, and cell genetic studies in glycogenosis type II","volume":"30","author":"Reuser","year":"1978","journal-title":"Am. J. Hum. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/S1474-4422(21)00001-6","article-title":"Onasemnogene abeparvovec gene therapy for symptomatic infantile-onset spinal muscular atrophy in patients with two copies of SMN2 (STR1VE): An open-label, single-arm, multicentre, phase 3 trial","volume":"20","author":"Day","year":"2021","journal-title":"Lancet Neurol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1038\/s41434-023-00402-4","article-title":"Approval and therapeutic value of gene therapies in the US and Europe","volume":"30","author":"Vokinger","year":"2023","journal-title":"Gene Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1089\/hum.2021.287","article-title":"Global Seroprevalence of Preexisting Immunity Against AAV5 and Other AAV Serotypes in People with Hemophilia A","volume":"33","author":"Klamroth","year":"2022","journal-title":"Hum. Gene Ther."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1089\/hum.2018.098","article-title":"Age-Related Seroprevalence of Antibodies Against AAV-LK03 in a UK Population Cohort","volume":"30","author":"Perocheau","year":"2019","journal-title":"Hum. Gene Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1586","DOI":"10.1128\/CVI.05107-11","article-title":"Adeno-associated virus antibody profiles in newborns, children, and adolescents","volume":"18","author":"Calcedo","year":"2011","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1089\/hum.2009.182","article-title":"Prevalence of serum IgG and neutralizing factors against adeno-associated virus (AAV) types 1, 2, 5, 6, 8, and 9 in the healthy population: Implications for gene therapy using AAV vectors","volume":"21","author":"Boutin","year":"2010","journal-title":"Hum. Gene Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1038\/gt.2014.47","article-title":"Neutralizing antibodies against AAV2, AAV5 and AAV8 in healthy and HIV-1-infected subjects in China: Implications for gene therapy using AAV vectors","volume":"21","author":"Liu","year":"2014","journal-title":"Gene Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"864","DOI":"10.1038\/s41598-020-57893-z","article-title":"Capsid-specific removal of circulating antibodies to adeno-associated virus vectors","volume":"10","author":"Bertin","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2084","DOI":"10.1038\/mt.2011.108","article-title":"A 10 patient case report on the impact of plasmapheresis upon neutralizing factors against adeno-associated virus (AAV) types 1, 2, 6, and 8","volume":"19","author":"Monteilhet","year":"2011","journal-title":"Mol. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"857276","DOI":"10.3389\/fimmu.2022.857276","article-title":"Overcoming the Challenges Imposed by Humoral Immunity to AAV Vectors to Achieve Safe and Efficient Gene Transfer in Seropositive Patients","volume":"13","author":"Gross","year":"2022","journal-title":"Front. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Unnisa, Z., Yoon, J.K., Schindler, J.W., Mason, C., and van Til, N.P. (2022). Gene Therapy Developments for Pompe Disease. Biomedicines, 10.","DOI":"10.3390\/biomedicines10020302"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3512","DOI":"10.1016\/j.ymthe.2021.08.020","article-title":"Cell type-selective targeted delivery of a recombinant lysosomal enzyme for enzyme therapies","volume":"29","author":"Baik","year":"2021","journal-title":"Mol. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1094279","DOI":"10.3389\/fimmu.2023.1094279","article-title":"Immune transgene-dependent myocarditis in macaques after systemic administration of adeno-associated virus expressing human acid alpha-glucosidase","volume":"14","author":"Hordeaux","year":"2023","journal-title":"Front. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1983","DOI":"10.1038\/mt.2010.175","article-title":"Preexisting immunity and low expression in primates highlight translational challenges for liver-directed AAV8-mediated gene therapy","volume":"18","author":"Hurlbut","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"675897","DOI":"10.3389\/fimmu.2021.675897","article-title":"Challenges Posed by Immune Responses to AAV Vectors: Addressing Root Causes","volume":"12","author":"Hamilton","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1089\/hum.2023.018","article-title":"Complement Activation by Adeno-Associated Virus-Neutralizing Antibody Complexes","volume":"34","author":"West","year":"2023","journal-title":"Hum. Gene Ther."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"999021","DOI":"10.3389\/fimmu.2022.999021","article-title":"Preexisting humoral immunity and complement pathway contribute to immunogenicity of adeno-associated virus (AAV) vector in human blood","volume":"13","author":"Smith","year":"2022","journal-title":"Front. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1089\/humc.2013.208","article-title":"Feasibility and safety of systemic rAAV9-hNAGLU delivery for treating mucopolysaccharidosis IIIB: Toxicology, biodistribution, and immunological assessments in primates","volume":"25","author":"Murrey","year":"2014","journal-title":"Hum. Gene Ther. Clin. 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