Gene edited B cells to co-express CNS-targeted antibodies

基因编辑 B 细胞共表达 CNS 靶向抗体

基本信息

批准号：
10475527
负责人：
Paula M Cannon
金额：
$ 20.63万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10475527
关键词：
Anti-Retroviral Agents Antibodies Antigens Antiviral Agents B cell therapy B-Cell Antigen Receptor B-Lymphocytes Benefits and Risks Biological Assay Blood Blood - brain barrier anatomy Blood Circulation Brain CRISPR/Cas technology Cells Central Nervous System Infections Choristoma Engineering Excision Exclusion Gene Transduction Agent Genes HIV HIV Antibodies HIV Infections Half-Life Human IgG1 Immune system Immunotherapy In Vitro Individual Liver Memory Modeling Mus Muscle Nerve Degeneration Penetrance Pharmaceutical Preparations Physiological Plasma Cells Property Protein Isoforms Proteins Recombinant Proteins Recombinants Regulation Resistance Series Serum Site Structure Surface Testing Tissues Vaccination Viral Viral reservoir Virus adeno-associated viral vector antibody engineering antiretroviral therapy base base editing cellular targeting design env Gene Products expression vector in vitro Assay interest mouse model nanobodies neutralizing antibody prevent receptor response transcytosis vaccination protocol vaccination strategy vector

项目摘要

ABSTRACT The CNS is a viral reservoir in HIV-infected individuals, 50% of whom will eventually develop HIV- associated neurodegeneration (HAND), despite antiretroviral drugs. Antibodies can also be powerful antivirals, promoting virus neutralization and the removal of HIV-infected cells through mechanisms such as ADCC and ADCP. Of special interest are broadly neutralizing antibodies (bnAbs), which can recognize the Env proteins from diverse viral strains and are therefore somewhat resistant to viral escape. As such, bnAbs are considered a promising approach for treating or preventing HIV infection, although the lack of induction of bnAbs by vaccination means that they are currently only used as injected proteins, or expressed from gene therapy vectors in non-immune tissues. An additional concern for CNS infections is that antibodies are present at lower rates in the CNS than the blood, although this can be enhanced by the addition of brain penetrating modules to the antibody. We have developed a gene editing strategy that allows us to reprogram human B cells to express specific antibodies. Our strategy takes advantage of the ability of B cells to respond to the presence of their cognate antigen, resulting in long-term secretion of the engineered antibody. We will now adapt this strategy to allow the co-expression of both bnAbs and modified brain-penetrating antibodies (BP-Ab) from the same edited cells. Using a range of assays, we will evaluate whether BP-Abs expressed from edited B cells retain full anti-HIV activities and show enhanced delivery to the CNS. In this way we will have developed the necessary proof of concept that a B cell therapy could be developed to provide long-term, HIV-responsive expression of brain- penetrating bnAbs.

抽象的中枢神经系统是 HIV 感染者的病毒库，其中 50% 最终会感染 HIV 尽管使用了抗逆转录病毒药物，但仍存在相关神经变性（HAND）。抗体也可以是强大的抗病毒药物，通过 ADCC 等机制促进病毒中和和清除 HIV 感染细胞 ADCP。特别令人感兴趣的是广泛中和抗体 (bnAbs)，它可以识别 Env 蛋白来自不同的病毒株，因此对病毒逃逸有一定的抵抗力。因此，bnAb 被认为是尽管缺乏 bnAb 的诱导，但这是一种治疗或预防 HIV 感染的有前途的方法疫苗接种意味着它们目前仅用作注射蛋白，或从基因治疗载体表达在非免疫组织中。对中枢神经系统感染的另一个担忧是，抗体在人体内的存在率较低。中枢神经系统比血液更重要，尽管这可以通过在中枢神经系统中添加脑穿透模块来增强抗体。我们开发了一种基因编辑策略，使我们能够重新编程人类 B 细胞以表达特定的抗体。我们的策略利用了 B 细胞对其同源细胞的存在做出反应的能力抗原，导致工程抗体的长期分泌。我们现在将调整此策略以允许来自相同编辑细胞的 bnAb 和修饰的脑穿透抗体 (BP-Ab) 共表达。通过一系列检测，我们将评估编辑过的 B 细胞表达的 BP-Ab 是否保留了完全的抗 HIV 能力活动并显示出对 CNS 的增强传递。通过这种方式，我们将开发出必要的证明一个概念是，可以开发 B 细胞疗法来提供长期的、对 HIV 敏感的大脑表达。穿透性bnAb。