AAV Induced Toxicity in the Eye

AAV 引起的眼部毒性

• 批准号: 10596582
• 负责人: CONSTANCE L CEPKO
• 金额: $ 41.64万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10596582
• 关键词: Animal Model; Animals; Biological Assay; Brain; Canis familiaris; Capsid; Cells; Cerebral cortex; Cessation of life; Clinical Trials; Cone; Disease; Dose; Eye; Eye diseases; Family suidae; Gene Delivery; Gene Expression; Genes; Genetic; Genetic Transcription; Genome; Genomics; Health; Infection; Inflammatory; Injections; Introns; Lead; Mediating; Mediator; Methods; Microglia; Morphology; Mus; Natural Immunity; Pathway interactions; Patients; Phenotype; Preparation; RNA; Retina; Retinal Cone; Retinal Detachment; Safety; Site; Source; Structure; Structure of retinal pigment epithelium; Testing; Therapeutic; Time; Tissues; Toxic effect; Viral; Virus; adeno-associated viral vector; cell type; gene therapy; inhibitor; loss of function; mouse model; nonhuman primate; pharmacologic; promoter; response; retinal neuron; subretinal injection; vector; viral detection

PROJECT SUMMARY / ABSTRACT AAV vectors have emerged as the leading vector for gene delivery to multiple tissues, proving to be both safe and efficacious in several clinical trials. However, they have not been fully explored for the limits of their safety and efficiency. As the therapeutic benefits of AAV mediated gene therapy will almost always increase with transduction of a greater number of cells, the safe delivery of high viral doses will likely provide a greater benefit to patients. However, high AAV doses in non-human primates (NHPs) and in other animal models have been associated with toxicity. If the mechanisms of these virus-induced problems can be elucidated, it may be possible to avoid them, so that a greater virus dose can be safely delivered. A mouse model allows for studies of mechanism that can then be investigated in larger animals. We have found that AAV vectors can harm ocular cells, in particular, cone photoreceptor cells and the retinal pigment epithelium (RPE) in mice, pigs, and dogs. This toxicity can lead to loss of retinal neurons and the RPE. Toxicity does not correlate with capsid type, the “cleanliness” of the stock, the gene that is expressed, or the preparation method. Rather, toxicity tracks with dose and genome sequence. We propose to identify the sequences that cause, and/or protect, against this toxicity. We also plan to track the cellular response to these toxic genomic sequences, to identify the cell type(s) in which toxicity is initiated, as well as the cell types which may amplify the results of viral detection. To this end, we will explore the RNA changes in several ocular cell types over time following infection with toxic and non-toxic AAV preparations. We will also follow the mechanism(s) that are triggered by these gene expression changes, and seek ways to block them. These findings will be extended to the brain, using injections of toxic and non-toxic stocks into the cerebral cortex. We will assay changes in brain RNA and cell health, using as probes the changes that we find in the retina. We will also test whether mechanisms that alleviate toxicity in the retina and in the brain.

项目摘要 /摘要 AAV矢量已成为多次基因传递的主要向量，因此既安全又安全 在几项临床试验中有效。但是，他们的安全限制尚未得到充分探索 和效率。由于AAV介导的基因疗法的治疗益处几乎总是随着 更大数量的细胞转导，高病毒剂量的安全输送可能会提供更大的 对患者的好处。但是，非人类灵长类动物（NHP）和其他动物模型中的高AAV剂量具有 与毒性有关。如果可以阐明这些病毒引起的问题的机制，则可能是 可以避免它们，以便可以安全地提供更大的病毒剂量。鼠标模型允许研究 然后可以在大型动物中研究的机制。我们发现AAV矢量会伤害 眼细胞，尤其是小鼠，猪和小鼠，猪和视网膜色素上皮（RPE）的锥细胞和视网膜色素上皮（RPE） 狗。这种毒性会导致残留神经元和RPE的丧失。毒性与衣壳类型无关， 股票的“清洁”，表达的基因或制备方法。相反，毒性轨迹 剂量和基因组序列。我们建议确定引起和/或保护的序列免受 这种毒性。我们还计划跟踪细胞对这些有毒基因组序列的反应，以鉴定细胞 引发毒性的类型以及可能扩大病毒检测结果的细胞类型。到 这最后，我们将探索有毒感染后随时间推移的几种眼细胞类型的RNA变化 和无毒的AAV制剂。我们还将遵循这些基因触发的机制 表达会改变，并寻求阻止它们的方法。这些发现将扩展到大脑 注射有毒和无毒的股票对大脑皮层的注射。我们将测定脑RNA和细胞的变化 健康，作为问题，我们在视网膜中发现的变化。我们还将测试是否有机制 减轻视网膜和大脑中的毒性。