Biodistribution and Shedding of AAV Vectors
Biodistribution and Shedding of AAV Vectors
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DOI：
10.1007/978-1-61779-370-7_15
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发表时间：
2011-01-01
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影响因子：
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通讯作者：
Arruda, Valder R.
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文献类型：
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作者：
Le Guiner, Caroline;Moullier, Phillipe;Arruda, Valder R.
确定AAV载体的生物分布和脱落对于所提议的早期临床试验的安全性评估至关重要。这在AAV载体的情况下尤其重要，因为它们被直接原位注射而不可能有中间离体步骤，例如在逆转录病毒介导的方法中。这种唯一的施用模式、高衣壳多样性（天然的和嵌合的）、各种递送途径（例如，肌内、静脉内、动脉内和颅内）使得生物分布和脱落研究成为未来几年的主要研究领域。事实上，只要工程衣壳，治疗策略，（cDNA相对于用于外显子跳跃的寡核苷酸的表达），给药方式继续快速发展为临床转化策略。生物分布和散毒研究的一个重要方面是，它们实际上不应该在“研究”中进行。模式，而是在监管动物药理学和毒理学研究的框架内，以便直接实施新药临床试验（IND）申请。然而，如果在早期研究阶段探索动物模型中的生物分布和脱落，为了研究以给定方式给药的给定AAV血清型是否转导某些免疫活性细胞（载体如何在免疫系统中自身分布以及以何种动力学分布？），建议使用类似于最终在临床阶段所做的生产和质量控制的AAV载体。2本章提供了研究AAV载体在给药后如何分布和脱落的方案和建议。我们讨论了（1）严格方法学的要求;（2）避免核酸交叉污染;（3）系统地评估检测灵敏度、特异性和重现性，因为环境可能截然不同，即，粪便与尿液;以及（4）在预期监管药理学/毒理学研究时选择适当的动物模型。
Determining the AAV vector biodistribution and shedding is central for the safety assessment of proposed early-phase clinical trials. It is especially crucial in the case of AAV vectors since they are injected directly in situ with no possibility of an intermediate ex vivo step, such as in retroviral-mediated approaches. This sole administration mode, the high capsid diversity (natural and chimeric), the various routes of delivery (e.g., intramuscular, intravenous, intra-arterial, and intracranial) make biodistribution and shedding studies a major investigational field for several years ahead. Indeed, the ideal scenario whereby they become generic is less likely to occur as long as the engineered capsid, the therapeutic strategies (expression of cDNA versus oligonucleotides for exon skipping), and the mode of delivery continue to evolve quickly to clinical translational strategies.An important aspect of biodistribution and shedding studies is that they practically should not be performed on a "research" mode but rather within the frame of the regulatory animal pharmacology and toxicology studies in order to directly implement the Investigational New Drug (IND) application. Yet, if biodistribution and shedding in animal models are explored at an early research stage, i.e., to investigate whether a given AAV serotype administered in a given way transduces certain immunocompetent cells (how does the vector distribute itself in the immune system and with what kinetic?), it is advisable to use an AAV vector manufactured and quality controlled similarly to what will be done ultimately at the clinical stage.This chapter provides protocols and recommendations to study how an AAV vector distributes and sheds after administration. We discuss (1) the requirements for a rigorous methodology; (2) avoiding nucleic acid cross contamination; (3) systematically assessing the assay sensitivity, specificity, and reproducibility because milieus can be drastically different, i.e., feces versus urine; and (4) choosing the appropriate animal model(s) when anticipating the regulatory pharmacological/toxicological studies.