Pre-Existing Immunity to a Nucleic Acid Contaminant-Derived Antigen Mediates Transaminitis and Resultant Diminished Transgene Expression in a Mouse Model of Hepatic Recombinant Adeno-Associated Virus-Mediated Gene Transfer.

IF 3.9 3区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Human gene therapy Pub Date : 2024-07-01 Epub Date: 2024-03-28 DOI:10.1089/hum.2023.188
Mark A Brimble, Christopher L Morton, Stephen M Winston, Isaiah L Reeves, Yunyu Spence, Pei-Hsin Cheng, Junfang Zhou, Amit C Nathwani, Paul G Thomas, Aisha Souquette, Andrew M Davidoff
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Abstract

Liver injury with concomitant loss of therapeutic transgene expression can be a clinical sequela of systemic administration of recombinant adeno-associated virus (rAAV) when used for gene therapy, and a significant barrier to treatment efficacy. Despite this, it has been difficult to replicate this phenotype in preclinical models, thereby limiting the field's ability to systematically investigate underlying biological mechanisms and develop interventions. Prior animal models have focused on capsid and transgene-related immunogenicity, but the impact of concurrently present nontransgene or vector antigens on therapeutic efficacy, such as those derived from contaminating nucleic acids within rAAV preps, has yet to be investigated. In this study, using Ad5-CMV_GFP-immunized immunocompetent BALB/cJ mice, and a coagulation factor VIII expressing rAAV preparation that contains green flourescent protein (GFP) cDNA packaged as P5-associated contaminants, we establish a model to induce transaminitis and observe concomitant therapeutic efficacy reduction after rAAV administration. We observed strong epitope-specific anti-GFP responses in splenic CD8+ T cells when GFP cDNA was delivered as a P5-associated contaminant of rAAV, which coincided and correlated with alanine and aspartate aminotransferase elevations. Furthermore, we report a significant reduction in detectable circulating FVIII protein, as compared with control mice. Lastly, we observed an elevation in the detection of AAV8 capsid-specific T cells when GFP was delivered either as a contaminant or transgene to Ad5-CMV_GFP-immunized mice. We present this model as a potential tool to study the underlying biology of post-AAV hepatotoxicity and demonstrate the potential for T cell responses against proteins produced from AAV encapsidated nontherapeutic nucleic acids, to interfere with efficacious gene transfer.

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在小鼠肝脏 rAAV 介导的基因转移模型中,对核酸污染物衍生抗原的预先免疫介导了转氨酶炎和转基因表达的减少。
在用于基因治疗的 rAAV 全身给药过程中,肝脏损伤并伴随治疗性转基因表达的缺失可能是一种临床后遗症,也是影响治疗效果的一个重要障碍。尽管如此,临床前模型一直难以复制这种表型,从而限制了该领域系统研究潜在生物机制和开发干预措施的能力。之前的动物模型主要关注的是囊膜和转基因相关的免疫原性,但同时存在的非转基因或载体抗原对疗效的影响(如来自 rAAV 预制品中污染核酸的抗原)仍有待研究。在这里,我们利用 Ad5-CMV_GFP 免疫BALB/cJ小鼠和含有作为P5相关污染物包装的GFP cDNA的凝血因子VIII表达rAAV制剂,建立了一个诱导转氨酶炎的模型,并观察了rAAV给药后随之而来的疗效降低。当 GFP cDNA 作为 rAAV 的 P5 相关污染物递送时,我们在脾脏 CD8+ T 细胞中观察到了强烈的表位特异性抗 GFP 反应,这种反应与丙氨酸和天门冬氨酸氨基转移酶升高相吻合和相关。此外,我们还发现,与对照组小鼠相比,可检测到的循环 FVIII 蛋白明显减少。最后,当 GFP 作为杂质或转基因传递给 Ad5-CMV_GFP 免疫小鼠时,我们观察到 AAV8 包膜特异性 T 细胞的检测率升高。我们将这一模型作为研究 AAV 后肝毒性的潜在生物学工具,并证明了针对由 AAV 封装的非治疗性核酸产生的蛋白质的 T 细胞反应有可能干扰有效的基因转移。
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来源期刊
Human gene therapy
Human gene therapy 医学-生物工程与应用微生物
CiteScore
6.50
自引率
4.80%
发文量
131
审稿时长
4-8 weeks
期刊介绍: Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases.
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