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

摘要 CNS是HIV感染者的病毒储存库，其中50%最终会发展为HIV-1。 相关的神经变性（手），尽管抗逆转录病毒药物。抗体也可以是强大的抗病毒药物， 通过ADCC等机制促进病毒中和和清除HIV感染细胞， ADCP。特别感兴趣的是广泛中和抗体（bnAb），其可以识别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，认为bnAb 这是一种有希望的治疗或预防HIV感染的方法，尽管缺乏通过 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。中枢神经系统感染的另一个问题是，在中枢神经系统中， 中枢神经系统比血液，虽然这可以通过增加大脑穿透模块来增强， 抗体的 我们已经开发出一种基因编辑策略，使我们能够重新编程人类B细胞， 抗体的我们的策略利用了B细胞对其同源物的存在做出反应的能力 抗原，导致工程化抗体的长期分泌。我们现在将调整这一战略， bnAb和修饰的脑穿透抗体（BP-Ab）两者从相同编辑的细胞共表达。 我们将使用一系列的检测方法来评估从编辑后的B细胞表达的BP-Ab是否保留了完整的抗HIV抗体 活性，并显示增强的CNS递送。这样，我们就可以证明 这一概念，B细胞疗法可以开发，以提供长期的，艾滋病毒反应性表达的脑- 穿透性bnAb。

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.