Quantifying the fitness cost of HIV-1 resistance to broadly neutralizing antibodies

项目总结/摘要： 用单克隆抗体（mAb）的免疫疗法已经在诸如自身免疫和免疫性疾病的环境中取得成功。 癌症，因此，使用抗体的治疗选择，特别是基于广泛 中和抗体（bNAb）应在HIV-1的背景下进行研究。一个警告是，HIV-1有能力 通过在其可变的env基因中产生突变来快速逃避抗体。因此迫切 需要深入了解HIV-1从bNAb逃逸，以帮助更有效的组合抗体策略， 用于HIV-1的治疗、治愈和预防。由于bNAb已经在临床试验中进行了测试， 最佳抗体组合不仅要评估它们的中和能力， 它们被不同的病毒“轻易逃脱”。本提案的总体目标是衡量 基因多样的HIV病毒株，以逃避广泛中和抗体，并设计一种抗体鸡尾酒， 能够限制逃跑。为了实现这一目标，Lynch博士是HIV-1从bNAbs中逃逸的专家。 已经组建了一个合作团队，使用分子病毒学（林奇博士）， 计算方法（巴顿和费舍尔博士）和人源化小鼠模型（克莱因博士）研究HIV-1 逃跑我们的中心假设是，当bNAb与HIV-1结合时，HIV-1从bNAb中逃逸将受到限制。 逃避所有bNAb所需的突变在不同的bNAb中发挥最大的复制适应性成本。 因此，复制不能通过补偿突变轻易恢复。我们将 通过（i）定义不同病毒中单个bNAb的可行逃逸途径的文库， 体外和体内方法，（ii）确定病毒-bNAb配对的适应性成本或逃逸的“容易性” 生物信息学，和（iii）确定抗体的最佳组合，最大限度地提高健身成本的 在不同的HIV-1亚型的耐药性，并在体外和体内测试这种组合。我们的结果将是 确定最佳组合抗体鸡尾酒，以限制各种HIV-1病毒逃逸的能力 抗体压力这些发现将为所有使用bNAb或基于bNAb的分子的临床试验提供信息， 最终，这些研究将确定未来表征抗体逃逸途径的合理途径。

PROJECT SUMMARY/ABSTRACT: Immunotherapy with monoclonal antibodies (mAbs) has been successful in settings such as autoimmunity and cancers, and therefore, treatment options with antibodies especially re-designed molecules based on broadly neutralizing antibodies (bNAbs) should be explored in the context of HIV-1. A caveat is that HIV-1 has the ability to rapidly escape from antibodies by generating mutations in its variable env gene. Therefore, there is an urgent need to gain insight into HIV-1 escape from bNAbs to aid in more effective combination antibody strategies to be used towards HIV-1 therapy, cure and prevention. As bNAbs are already being tested in clinical trials, it is imperative that optimal antibody combinations are evaluated not only for their neutralization capability but also their ‘ease of escape’ by diverse viruses. The overall objective of this proposal is to measure the ability of genetically diverse strains of HIV to escape from broadly neutralizing antibodies and design an antibody cocktail capable of restricting escape. To accomplish this goal, Dr. Lynch, who is an expert in HIV-1 escape from bNAbs has put together a collaborative team for an interdisciplinary approach using molecular virology (Dr. Lynch), computational methods (Drs. Barton and Fischer) and a humanized-mouse model (Dr. Klein) to study HIV-1 escape. Our central hypothesis is that HIV-1 escape from combination bNAbs will be limited when the mutations required to escape all bNAbs exert the maximum replicative fitness cost across diverse viruses and, therefore, replication cannot easily be restored through compensatory mutations. We will test this hypothesis by (i) defining a library of viable escape pathways for single bNAbs in diverse viruses with in vitro and in vivo approaches, (ii) determining the fitness cost or “ease” of escape for virus-bNAb pairings bioinformatically, and (iii) identifying the optimal combination of antibodies that maximize fitness costs of resistance in diverse HIV-1 subtypes and testing this combination in vitro and in vivo. Our outcome will be the identification of optimal combination antibody cocktails to limit the ability of diverse HIV-1 viruses to escape from antibody pressure. These findings will inform all clinical trials using bNAbs or bNAb-based molecules, and ultimately, these studies will define a rational pipeline to characterize antibody escape pathways in the future.