{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T23:42:05Z","timestamp":1790811725948,"version":"4.1.0"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,3,22]],"date-time":"2023-03-22T00:00:00Z","timestamp":1679443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Immune responses to human non-self transgenes can present challenges in preclinical studies of adeno-associated virus (AAV) gene therapy candidates in nonhuman primates. Although anti-transgene immune responses are usually mild and non-adverse, they can confound pharmacological readouts and complicate translation of results between species. We developed a gene therapy candidate for Pompe disease consisting of AAVhu68, a clade F AAV closely related to AAV9, that expresses an engineered human acid-alpha glucosidase (hGAA) tagged with an insulin-like growth factor 2 variant (vIGF2) peptide for enhanced cell uptake. Rhesus macaques were administered an intravenous dose of 1x10<jats:sup>13<\/jats:sup> genome copies (GC)\/kg, 5x10<jats:sup>13<\/jats:sup> GC\/kg, or 1 x 10<jats:sup>14<\/jats:sup> GC\/kg of AAVhu68.vIGF2.hGAA. Some unusually severe adaptive immune responses to hGAA presented, albeit with a high degree of variability between animals. Anti-hGAA responses ranged from absent to severe cytotoxic T-cell-mediated myocarditis with elevated troponin I levels. Cardiac toxicity was not dose dependent and affected five out of eleven animals. Upon further investigation, we identified an association between toxicity and a major histocompatibility complex class I haplotype (Mamu-A002.01) in three of these animals. An immunodominant peptide located in the C-terminal region of hGAA was subsequently identified <jats:italic>via<\/jats:italic> enzyme-linked immunospot epitope mapping. Another notable observation in this preclinical safety study cohort pertained to the achievement of robust and safe gene transfer upon intravenous administration of 5x10<jats:sup>13<\/jats:sup> GC\/kg in one animal with a low pre-existing neutralizing anti-capsid antibodies titer (1:20). Collectively, these findings may have significant implications for gene therapy inclusion criteria.<\/jats:p>","DOI":"10.3389\/fimmu.2023.1094279","type":"journal-article","created":{"date-parts":[[2023,3,22]],"date-time":"2023-03-22T05:50:30Z","timestamp":1679464230000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":25,"title":["Immune transgene-dependent myocarditis in macaques after systemic administration of adeno-associated virus expressing human acid alpha-glucosidase"],"prefix":"10.3389","volume":"14","author":[{"given":"Juliette","family":"Hordeaux","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali","family":"Ramezani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steve","family":"Tuske","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nickita","family":"Mehta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunjuan","family":"Song","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Lynch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherine","family":"Lupino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica A.","family":"Chichester","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth L.","family":"Buza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cecilia","family":"Dyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongwei","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Bell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jill M.","family":"Weimer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hung","family":"Do","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James M.","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,3,22]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1038\/nm1358","article-title":"Successful transduction of liver in hemophilia by AAV-factor IX and limitations imposed by the host immune response","volume":"12","author":"Manno","year":"2006","journal-title":"Nat Med"},{"key":"B2","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1182\/blood-2013-01-306647","article-title":"Immune responses to AAV vectors: Overcoming barriers to successful gene therapy","volume":"122","author":"Mingozzi","year":"2013","journal-title":"Blood"},{"key":"B3","doi-asserted-by":"publisher","first-page":"797","DOI":"10.1089\/hum.2013.092","article-title":"Anti-dystrophin T cell responses in duchenne muscular dystrophy: Prevalence and a glucocorticoid treatment effect","volume":"24","author":"Flanigan","year":"2013","journal-title":"Hum Gene Ther"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1000228","article-title":"Dystrophin immunity in duchenne's muscular dystrophy","volume":"363","author":"Mendell","year":"2010","journal-title":"N Engl J Med"},{"key":"B5","first-page":"3","article-title":"Collaborative analysis by clinical trial sponsors and academic experts of anti-transgene SEAS in studies of gene therapy for DMD","volume":"30","author":"Bonnemann","year":"2022","journal-title":"Mol Ther"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1086\/595830","article-title":"Worldwide epidemiology of neutralizing antibodies to adeno-associated viruses","volume":"199","author":"Calcedo","year":"2009","journal-title":"J Infect Dis"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2019.012","article-title":"Safe and sustained expression of human iduronidase after intrathecal administration of adeno-associated virus serotype 9 in infant rhesus monkeys","volume":"30","author":"Hordeaux","year":"2019","journal-title":"Hum Gene Ther"},{"key":"B8","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1016\/j.omtm.2018.06.004","article-title":"Toxicology study of intra-cisterna magna adeno-associated virus 9 expressing iduronate-2-Sulfatase in rhesus macaques","volume":"10","author":"Hordeaux","year":"2018","journal-title":"Mol Ther Methods Clin Dev"},{"key":"B9","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/j.omtm.2018.06.003","article-title":"Toxicology study of intra-cisterna magna adeno-associated virus 9 expressing human alpha-L-Iduronidase in rhesus macaques","volume":"10","author":"Hordeaux","year":"2018","journal-title":"Mol Ther Methods Clin Dev"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2007-03-080093","article-title":"Modulation of tolerance to the transgene product in a nonhuman primate model of AAV-mediated gene transfer to liver","volume":"110","author":"Mingozzi","year":"2007","journal-title":"Blood"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2009.060","article-title":"Adeno-associated virus-mediated gene transfer to nonhuman primate liver can elicit destructive transgene-specific T cell responses","volume":"20","author":"Gao","year":"2009","journal-title":"Hum Gene Ther"},{"key":"B12","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1089\/humc.2017.014","article-title":"Non-clinical study examining AAV8.Tbg.Hldlr vector-associated toxicity in chow-fed wild-type and ldlr(+\/-) rhesus macaques","volume":"28","author":"Greig","year":"2017","journal-title":"Hum Gene Ther Clin Dev"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.15252\/emmm.202113968","article-title":"Muscle-directed gene therapy corrects pompe disease and uncovers species-specific GAA immunogenicity","volume":"14","author":"Eggers","year":"2022","journal-title":"EMBO Mol Med"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1177\/0192623316677326","article-title":"Identification of MHC haplotypes associated with drug-induced hypersensitivity reactions in cynomolgus monkeys","volume":"45","author":"Wu","year":"2017","journal-title":"Toxicol Pathol"},{"key":"B15","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1186\/s13023-017-0693-2","article-title":"A study on the safety and efficacy of reveglucosidase Alfa in patients with late-onset pompe disease","volume":"12","author":"Byrne","year":"2017","journal-title":"Orphanet J Rare Dis"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.18609\/cgti.2021.207","article-title":"No cell left behind: Engineering gene therapies for cross correction","volume":"7","author":"Weimer JMB","year":"2021","journal-title":"Cell Gene Ther Insights"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0902534106","article-title":"Neural deficits contribute to respiratory insufficiency in pompe disease","volume":"106","author":"DeRuisseau","year":"2009","journal-title":"Proc Natl Acad Sci U.S.A."},{"key":"B18","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1186\/s40478-017-0464-2","article-title":"Long-term neurologic and cardiac correction by intrathecal gene therapy in pompe disease","volume":"5","author":"Hordeaux","year":"2017","journal-title":"Acta Neuropathol Commun"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2020.04.019","article-title":"Combined CNS and systemic directed gene therapy in a mouse model of pompe disease with advanced disease at treatment","volume":"28","author":"Hordeaux","year":"2020","journal-title":"Mol Ther"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.aba9188","article-title":"MicroRNA-mediated inhibition of transgene expression reduces dorsal root ganglion toxicity by AAV vectors in primates","volume":"12","author":"Hordeaux","year":"2020","journal-title":"Sci Transl Med"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2013.131","article-title":"Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers by droplet digital PCR","volume":"25","author":"Lock","year":"2014","journal-title":"Hum Gene Ther Methods"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2018.041","article-title":"Standardized method for intra-cisterna magna delivery under fluoroscopic guidance in nonhuman primates","volume":"29","author":"Katz","year":"2018","journal-title":"Hum Gene Ther Methods"},{"key":"B23","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1089\/hgtb.2018.038","article-title":"Assessment of humoral, innate, and T-cell immune responses to adeno-associated virus vectors","volume":"29","author":"Calcedo","year":"2018","journal-title":"Hum Gene Ther Methods"},{"key":"B24","doi-asserted-by":"publisher","first-page":"196","DOI":"10.1093\/ilar\/ilt036","article-title":"Haplessly hoping: Macaque major histocompatibility complex made easy","volume":"54","author":"Wiseman","year":"2013","journal-title":"ILAR J"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2020.167","article-title":"Adeno-associated virus-induced dorsal root ganglion pathology","volume":"31","author":"Hordeaux","year":"2020","journal-title":"Hum Gene Ther"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1172\/jci.insight.125358","article-title":"Improved efficacy of a next-generation ERT in murine pompe disease","volume":"4","author":"Xu","year":"2019","journal-title":"JCI Insight"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.2174\/156652307782151425","article-title":"Immune responses to AAV in clinical trials","volume":"7","author":"Mingozzi","year":"2007","journal-title":"Curr Gene Ther"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1617726114","article-title":"Class I-restricted T-cell responses to a polymorphic peptide in a gene therapy clinical trial for alpha-1-Antitrypsin deficiency","volume":"114","author":"Calcedo","year":"2017","journal-title":"Proc Natl Acad Sci U.S.A."},{"key":"B29","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.clim.2019.01.009","article-title":"HLA- and genotype-based risk assessment model to identify infantile onset pompe disease patients at high-risk of developing significant anti-drug antibodies (ADA)","volume":"200","author":"De Groot","year":"2019","journal-title":"Clin Immunol"},{"key":"B30","doi-asserted-by":"publisher","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"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1094279\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,3,22]],"date-time":"2023-03-22T05:50:34Z","timestamp":1679464234000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1094279\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,22]]},"references-count":30,"alternative-id":["10.3389\/fimmu.2023.1094279"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2023.1094279","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,22]]},"article-number":"1094279"}}