Leveraging CTLs targeting highly networked epitopes to suppress the latent HIV-1 reservoir

项目摘要/摘要 这项建议提出了一个五年的研究生涯发展计划，重点是研究对外汉语学习 作为包括在治疗性CTL中的一组新的不变靶点，对高度联网的表位的反应- 基于HIV-1的疫苗。候选人目前是哈佛医学院的医学讲师， 马萨诸塞州总医院的胃肠病科。概述的提案建立在 候选人之前在HIV-1免疫学和生物化学方面的研究经验，他在那里定义了CTL 带有结构和功能限制的表位，以及优先针对个人的表位 自然控制HIV-1病毒的人。现在，他在他的导师布鲁斯·沃克博士的指导下，在 以确定这些表位是否可能成为有价值的CTL靶标 在接受治疗的个体中。建议的实验和教学工作将使候选人 拥有一套独特的技能，使他能够过渡到独立，成为一名内科科学家，在 预防和治疗性HIV-1疫苗接种学。 艾滋病毒/艾滋病流行病继续在世界范围内产生巨大的医疗、社会和经济影响。 虽然联合抗逆转录病毒疗法(CART)帮助大大减轻了全球艾滋病毒的负担，但 病毒建立持久潜伏蓄水池的能力需要对艾滋病毒感染者进行终身治疗 个人。因此，可以抑制或消除病毒库从而限制艾滋病毒的新方法 治疗时间是非常必要的。最近的努力主要集中在诱导细胞毒性T细胞上 作为治疗性疫苗的潜在目标。然而，CTL逃逸突变在体内的积累 慢性感染的CART抑制患者限制了成功防止病毒反弹的能力 手推车停下来。在他的博士后研究期间，候选人开发了一种新的方法，称为 基于结构的网络分析，识别由广泛的HLA等位基因呈现的特定表位， 它们不能忍受突变。他还证明了以高度联网的CTL表位为靶点 能够区分自发控制HIV-1的人和患有进展性疾病的人。这个 候选人现在假设CTL介导的免疫反应是针对高度“网络化”的 在接受CART治疗的慢性感染患者中，表位也可以抑制CART停止后的病毒生长 个人。这一假设将通过以下目标得到验证：1)对 前病毒DNA中的高度网络化表位，2)评估针对高度网络化表位的CTL是否可以 抑制接受CART治疗的患者的病毒生长和3)发展编码 多个高度联网的表位，并评估其在体内诱导CTL反应的能力。有效的CTL- 通过基于结构的网络分析确定的高度网络化表位的介导性反应可能限制病毒 从潜伏感染的CD4+T细胞中反弹，从而可能指导治疗性CTL的合理设计 基于HIV-1的疫苗。

Project Summary/Abstract This proposal presents a five year research career development program focused on the study of CTL responses to highly “networked” epitopes as a new set of invariant targets to include in a therapeutic CTL- based vaccine for HIV-1. The candidate is currently an Instructor of Medicine at Harvard Medical School and the Division of Gastroenterology at Massachusetts General Hospital. The outlined proposal builds on the candidate's previous research experience in HIV-1 immunology and biochemistry where he defined CTL epitopes that carry structural and functional constraints, and which are preferentially targeted by individuals who naturally control HIV-1. He is now positioned, under the guidance of his mentor Dr. Bruce Walker at the Ragon Institute of MGH, MIT and Harvard, to determine whether these epitopes could be valuable CTL targets in treatment-suppressed individuals. The proposed experiments and didactic work will position the candidate with a unique set of skills that will enable him transition to independence as a physician-scientist in the field of prophylactic and therapeutic HIV-1 vaccinology. The HIV/AIDS epidemic continues to have enormous medical, societal and economic implications worldwide. While combination anti-retroviral therapy (cART) has helped to greatly reduce the global burden of HIV, the ability of the virus to establish a persistent latent reservoir requires lifelong treatment for HIV-infected individuals. As a result, new modalities that can suppress or eliminate the viral reservoir and thereby limit HIV treatment duration are greatly needed. Recent efforts have been focused on the induction of cytotoxic T cells as potential targets for therapeutic vaccines. However, the accumulation of CTL escape mutations in chronically infected cART-suppressed patients limits the ability to successfully prevent viral rebound following cART cessation. During his postdoctoral fellowship, the candidate developed a new approach known as structure-based network analysis that identifies specific epitopes, presented by a broad array of HLA alleles, which are intolerant to mutations. He also demonstrated that the targeting of highly “networked” CTL epitopes is able to distinguish individuals who spontaneously control HIV-1 from those with progressive disease. The candidate now hypothesizes that CTL mediated immune responses directed against highly “networked” epitopes can also suppress viral outgrowth following cART cessation in chronically infected cART-treated individuals. This hypothesis will be tested through the following aims: 1) Perform deep mutational scanning of highly networked epitopes in proviral DNA, 2) Assess whether CTLs targeting highly networked epitopes can suppress viral outgrowth from cART-treated patients and 3) Develop an adenovirus (Ad) vector encoding multiple highly networked epitopes and assess its ability to induce CTL responses in vivo. Effective CTL- mediated responses to highly networked epitopes identified by structure-based network analysis may limit viral rebound from latently infected CD4+ T cells and thereby may guide the rational design of a therapeutic CTL- based vaccine for HIV-1.