AAV Vectored Delivery of Broadly Neutralizing Antibodies with Optimal Innate Functionality Against HIV

项目摘要/摘要 本提案描述了为Alejandro Balazs博士提供R01赠款的框架。巴拉兹博士是 目前是哈佛医学院的助理教授，在Ragon担任首席研究员 麻省理工学院和哈佛大学麻省理工学院。巴拉兹博士的研究重点是通过基因改造免疫系统 将转移作为预防艾滋病毒的一种新手段。广谱中和抗体(BNAbs) 人类免疫缺陷病毒(HIV)在HIV的预防和治疗方面显示出巨大的希望，因为它们具有强大的 中和大量在全球流行的艾滋病毒毒株。多项动物实验和临床试验 试验表明，bNAbs具有抵抗病毒攻击和减少病毒血症的能力。 已确定的感染。BNAbs可通过阻断病毒附着或膜融合来抑制HIV感染； 然而，最近的工作表明，抗体的片段结晶区(Fc)也可能起到作用 通过与先天免疫的相互作用，对bNAb介导的HIV抑制作用具有重要意义。这项建议旨在 使用基于细胞的体外分析来确定下一代HIV bNAbs与效应器的结合程度 功能，如抗体依赖的细胞毒性(ADCC)和抗体依赖的细胞 吞噬作用(ADCP)。这项提案将修改由Dr。 Balazs可持续表达先前具有精确Fc区突变的抗体 证实能增强或消除Fc-受体的相互作用。通过操纵特定性和 这些抗体的浓度，这项研究将定义适用于Fc受体结合的规则 预防艾滋病病毒感染。此外，它还试图确定Fc增强的抗体对 提高bNAb预防艾滋病毒传播的效力。最后，这项提案将操纵 人源化小鼠免疫系统作为解剖和精确量化其贡献的手段 特异性免疫细胞可预防艾滋病毒传播。总而言之，这项工作将揭示最佳表位靶标 以及天然免疫机制，以产生编码具有增强的bNAbs的下一代AAV载体 先天免疫功能，防止艾滋病毒传播。

Project Summary / Abstract This proposal describes the framework of an R01 grant for Alejandro Balazs, PhD. Dr. Balazs is currently an assistant professor at Harvard Medical School working as a principal investigator at the Ragon Institute of MGH, MIT & Harvard. Dr. Balazs’ research is focused on engineering the immune system via gene transfer as a novel means of creating protection against HIV. Broadly neutralizing antibodies (bNAbs) against human immunodeficiency virus (HIV) show great promise in HIV prevention and therapy as they potently neutralize a significant breadth of globally circulating HIV strains. A number of animal experiments and clinical trials have demonstrated the ability of bNAbs to confer protection from viral challenge and reduce viremia of established infections. BNAbs can inhibit HIV infection by blocking viral attachment or membrane fusion; however, recent work suggests that the fragment crystallizable (Fc) region of antibodies may also contribute significantly to bNAb-mediated HIV inhibition through interactions with innate immunity. This proposal seeks to use in vitro cell-based assays to determine the extent to which next-generation HIV bNAbs engage effector functions, such as antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). This proposal will modify the Vectored ImmunoProphylaxis technology pioneered by Dr. Balazs to generate sustained expression of antibodies harboring precise Fc-region mutations previously demonstrated to enhance or abrogate, Fc-receptor interactions. By manipulating the specificity and concentration of these antibodies, this study will define the rules governing Fc-receptor engagement that apply to prevention of HIV acquisition. Furthermore, it seeks to determine the potential for Fc-enhanced antibodies to increase the potency of bNAb protection against HIV transmission. Finally, this proposal will manipulate the immune system of humanized mice as a means of dissecting and precisely quantifying the contribution of specific immune cells to prevention of HIV transmission. Together, this work will reveal optimal epitope targets and innate immune mechanisms to produce next-generation AAV vectors encoding bNAbs with enhanced innate immune function to prevent HIV transmission.