An AAV Approach to Treating HIV

治疗 HIV 的 AAV 方法

基本信息

批准号：
9292034
负责人：
Matthew Ryan Gardner
金额：
$ 5.92万
依托单位：
SCRIPPS FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9292034
关键词：
Address Affinity Animals Anti-Retroviral Agents Antibodies Antibody Therapy Antibody titer measurement Binding Sites Biological Assay CCR5 gene CXCR4 gene Cell surface Data Dependovirus Detection Disease Dose Fellowship Gene Transduction Agent Goals HIV HIV Entry Inhibitors HIV vaccine HIV-1 HIV-2 Human Immune response Individual Infection Injection of therapeutic agent Innate Immune Response Interferons Intravenous Macaca Macaca mulatta Measurable MicroRNAs Pathway interactions Pharmacotherapy Positioning Attribute Principal Investigator Production Proteins Publishing Regulation Research Research Training Resistance SIV Safety Testing Time Transgenes Unspecified or Sulfate Ion Sulfates Viral Viral Load result Viremia Virus Work World Health Organization adeno-associated viral vector antiretroviral therapy base experimental study gene therapy humanized mouse immunogenic improved in vitro Assay in vivo inhibitor/antagonist neutralizing antibody peptidomimetics receptor response simian human immunodeficiency virus transgene expression viral RNA viral rebound

项目摘要

PROJECT SUMMARY We have recently published our research that characterizes eCD4-Ig. eCD4-Ig is an antibody-like HIV entry inhibitor that fuses a sulfated CCR5-mimetic peptide to the C-terminus of CD4-Ig. Based on neutralization assay data, eCD4-Ig is broader than and equally potent as some of the best described HIV-1 broadly neutralizing antibodies to date. eCD4-Ig neutralized all isolates tested including 38 HIV-1 isolates resistant to 3BNC117 or NIH45-46, HIV-2, SIVmac239, SIVmac251, and isolates that use CXCR4 as their coreceptor. The addition of the CCR5-mimetic peptide allowed eCD4-Ig to have higher affinity for cell surface-expressed HIV-1 Env and also limited viral enhancement caused by sub-neutralizing levels of CD4-Ig. Using adeno-associated virus (AAV) vectors, we have shown that a rhesus form of eCD4-Ig can be expressed in four rhesus macaques for over one year with no harm to the animals. The rh- eCD4-Ig protein titers were at levels that protected all four macaques from multiple SHIV-AD8 challenges up to 16-times the AID50 (Animal Infectious Dose 50). We did observe a measurable anti- transgene response to the expressed rh-eCD4-Ig protein, but the response was not near the levels seen against AAV-delivered HIV-1 antibodies. Yet, even a relatively modest immune response to the delivered transgene can limit the inhibitor’s efficacy in vivo. With these encouraging data, this proposal seeks to answer two main questions: (1) Can AAV-expressed eCD4-Ig be used as a therapy to maintain viral suppression in SHIV infected macaques? (2) Can interferon-induced miRNAs regulate transgene expression from AAV vectors to limit the host immune response to the transgene? With the primary goal of using eCD4-Ig as an alternative to antiretroviral therapies, answering these two questions will continue to build on the safety and efficacy of AAV-delivered eCD4-Ig as well as realizing the complete potential of AAV vectors used for gene therapy.

项目概要我们最近发表的研究表明 eCD4-Ig 是一种类似 HIV 的抗体。将硫酸化 CCR5 模拟肽融合到 CD4-Ig C 末端的进入抑制剂。中和测定数据显示，eCD4-Ig 比一些最好描述的方法更广泛且同样有效迄今为止，HIV-1 广泛中和抗体 eCD4-Ig 中和了所有测试的分离株，包括 38 个 HIV-1。对 3BNC117 或 NIH45-46、HIV-2、SIVmac239、SIVmac251 具有抗性的分离株，以及使用 CXCR4 作为抗药性的分离株添加 CCR5 模拟肽使得 eCD4-Ig 对它们具有更高的亲和力。细胞表面表达的 HIV-1 Env 以及亚中和水平引起的有限病毒增强使用腺相关病毒 (AAV) 载体，我们发现了恒河猴形式的 eCD4-Ig。可以在四只恒河猴中表达一年多，并且对动物没有伤害。 eCD4-Ig 蛋白滴度达到保护所有四只猕猴免受多种 SHIV-AD8 感染的水平挑战高达 AID50（动物感染剂量 50）的 16 倍，我们确实观察到了可测量的抗- 转基因对表达的 rh-eCD4-Ig 蛋白有反应，但反应并未接近水平然而，即使对 AAV 传递的 HIV-1 抗体也有相对温和的免疫反应。递送的转基因可以限制抑制剂在体内的功效。根据这些令人鼓舞的数据，该提案提出了这一建议。试图回答两个主要问题：(1) AAV 表达的 eCD4-Ig 能否用作治疗在 SHIV 感染的猕猴中维持病毒抑制？ (2) 干扰素诱导的 miRNA 可以调节吗？ AAV 载体的转基因表达可限制宿主对转基因的免疫反应？使用 eCD4-Ig 作为抗逆转录病毒疗法替代方案的主要目标，回答了这两个问题问题将继续建立在 AAV 传递的 eCD4-Ig 的安全性和有效性以及实现 AAV 载体用于基因治疗的全部潜力。