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

• 批准号: 10589115
• 负责人: Paula M Cannon
• 金额: $ 24.75万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 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/10589115
• 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; Brain; CRISPR/Cas technology; Cells; Central Nervous System Infections; Choristoma; Circulation; Engineering; Engraftment; 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; Penetration; 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 editing; cellular targeting; design; env Gene Products; expression vector; in vitro Assay; interest; mouse model; nanobodies; neutralizing antibody; prevent; programs; receptor; response; transcytosis; vaccination protocol; vaccination strategy; vector

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.

摘要 中枢神经系统是艾滋病毒感染者的病毒库，其中50%的人最终将发展为艾滋病毒- 相关神经变性(手)，尽管有抗逆转录病毒药物。抗体也可以是强大的抗病毒药物， 通过ADCC和ADCC等机制促进病毒中和和清除艾滋病毒感染细胞 ADCP。特别令人感兴趣的是广谱中和抗体(BNAbs)，它可以识别环境蛋白 来自不同的病毒株，因此对病毒逃逸有一定的抵抗力。因此，考虑使用bNAbs 一种治疗或预防艾滋病毒感染的有希望的方法，尽管缺乏通过 疫苗接种意味着它们目前只被用作注射蛋白，或通过基因治疗载体表达 在非免疫组织中。中枢神经系统感染的另一个令人担忧的问题是， 中枢神经系统比血液，尽管这可以通过添加脑穿透模块 抗体。 我们已经开发出一种基因编辑策略，允许我们重新编程人类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。我们现在将调整这一策略，以允许 来自相同编辑细胞的bNAbs和修饰的脑穿透抗体(BP-Ab)的共表达。 通过一系列的检测，我们将评估从编辑的B细胞表达的BP-Abbs是否保留了完全的抗HIV 活动，并显示加强了对中枢神经系统的交付。通过这种方式，我们将开发出必要的证据 可以开发一种B细胞疗法来提供长期的、对艾滋病毒有反应的大脑表达-- 穿透性bNAbs。