The Effect of Natural Killer Cell-Based Therapy on the HIV Reservoir

项目摘要/摘要 这份拨款提案描述了乔斯林博士职业发展的五年指导培训计划。 金是一名内科科学家，他将开发尖端方法来量化艾滋病毒宿主并调查 治愈艾滋病毒感染的新疗法。她最近发现，自然杀伤(NK)细胞可以延迟病毒 在复杂的人性化小鼠模型中反弹的艾滋病毒潜伏期。她还发现，抗艾滋病毒嵌合体 NK细胞上的抗原受体(CAR)在体外有效地杀伤HIV感染的细胞。此外，一种新的合成潜伏期 再活化剂(LRA)在体内ART存在的情况下激活潜伏的HIV。重要的是，她创新了一种 基因条形码报告HIV-1，并证明了这个多样化的病毒群具有复制能力 和体内致病性，并形成一个潜伏的蓄水池，在停止抗逆转录病毒治疗后反弹 (艺术)。她将使用这种新的条形码病毒工具来准确量化潜伏感染的克隆的数量 在活体内。这些初步数据是当前提案的基础，其中涉及完成 遵循目标。首先，她将调查来自外周血或造血干细胞的NK细胞 使用干细胞的新方法，祖细胞(HSCP)在延缓HIV在体内反弹方面更有效 辨析。其次，她将测试CAR-NK细胞和CAR-T细胞在病毒反弹方面的有效性 在活体内。最后，她将在基因条码病毒中使用最先进的方法来确定 联合疗法(LRA和基于NK细胞的疗法)会影响体内的HIV储存库。在K08的资助下， 她我们将完成这些研究，目的是加深我们对艾滋病毒发病机制和 治疗策略。金博士目前是加州大学洛杉矶分校传染病部门的临床助理教授。 最近从加州大学洛杉矶分校的专业培训和高级研究(STAR)项目毕业，从那里她 获加州理工学院生物与生物工程专业博士学位 大卫·巴尔的摩医生顾问。在拟议的授勋期间，她将继续在主要导师的指导下工作。 加州大学洛杉矶分校的杰罗姆·扎克博士，由盖伊博士组成的咨询委员会提供额外的指导 克鲁克斯(加州大学洛杉矶分校)、亚历克斯·霍夫曼(加州大学洛杉矶分校)和朱迪思·柯里尔(加州大学洛杉矶分校)。在他们的指导下，她将实现 成功申请独立资助和过渡到 独立。因此，通过她的合作和说教培训，她将扩大她的NK细胞 计划并使用复杂的计算技能进行高通量条形码测序分析，以了解 艾滋病病毒携带者。她还将在干细胞分化和分析、分子克隆、 还有写作和手艺。K08的资助将使她能够发展成为 一位成功的独立调查员。

PROJECT SUMMARY/ABSTRACT This grant proposal describes a five-year mentored training program for the career development of Dr. Jocelyn Kim, a physician-scientist, who will develop cutting-edge methods to quantify the HIV reservoir and investigate novel therapies to cure HIV infection. She has recently discovered that natural killer (NK) cells delays viral rebound in a sophisticated humanized mouse model of HIV latency. She has also found that anti-HIV chimeric antigen receptor (CAR)-on NK cells efficiently kill HIV-infected cells in vitro. In addition, a novel synthetic latency reactivating agent (LRA) activates latent HIV in the presence of ART in vivo. Importantly, she has innovated a genetically barcoded reporter HIV-1 and demonstrated that this diverse virus swarm is replication-competent and pathogenic in vivo and forms a latent reservoir, which rebounds after cessation of antiretroviral therapy (ART). She will use this novel barcoded-virus tool to accurately quantify the number of latently infected clones in vivo. These preliminary data are the basis of the current proposal, which involves the completion of the following aims. First, she will investigate whether NK cells from peripheral blood or hematopoietic stem cell progenitors (HSCPs) are more effective in delaying HIV rebound in vivo using new methods of stem cell differentiation and analysis. Second, she will test of efficacy of CAR-NK cells versus CAR-T cells on viral rebound in vivo. Lastly, she will use state-of-the-art approach in genetic barcoded virus to determine whether combinatorial therapies (LRA and NK cell-based therapies) affect the HIV reservoir in vivo. With K08 funding, she we will complete these studies with the intention of deepening our understanding of HIV pathogenesis and curative strategies. Dr. Kim is currently an assistant clinical professor in the UCLA Division of Infectious Diseases and recent graduate of the UCLA Specialty Training and Advanced Research (STAR) program, from which she received a PhD in Biology and Biological Engineering from California Institute of Technology under her PhD advisor Dr. David Baltimore. During the proposed award period, she will continue under the primary mentorship of Dr. Jerome Zack at UCLA with additional mentorship through an advisory committee comprised of Drs. Gay Crooks (UCLA), Alex Hoffman (UCLA) and Judith Currier (UCLA). With their guidance, she will attain the necessary research and career skills to apply successfully for independent funding and transition to independence. As such, through her collaborations as well as didactic training, she will expand her NK cell project and use complex computational skills for high-throughout barcode sequencing analysis to understand the HIV reservoir. She will also acquire new skills in stem cell differentiation and analysis, molecular cloning, and writing and grantsmanship. K08 funding will allow her to develop all of the skills and tools needed to become a successful independent investigator.