Combination immunotherapy with gene modified CD4 and CD8 T cells and stem cells

基因修饰 CD4 和 CD8 T 细胞与干细胞的联合免疫疗法

• 批准号: 7659739
• 负责人: CARL H. JUNE
• 金额: $ 196.72万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7659739
• 关键词: Combination; immunotherapy; gene; modified; CD4

DESCRIPTION (provided by applicant): Our long term goal is to build an HIV-resistant immune system. Based prior studies in the field of the immunopathogenesis of HIV infection, it is likely that both enhanced HlV-specific immunity, as well as a population of CD4 cells that have enhanced resistance to HIV infection will be required to achieve this goal. In our ongoing IPCP grant (expiry February 2009), in collaboration with Sangamo Biosciences, we have developed HIV resistant CD4 T cells using zinc finger nucleases to knock out the CCR5 coreceptor, which is being tested in the clinic in 2009 (Perez et al, in press). To generate enhanced HlV-specific immunity, in collaboration with Adaptimmune Ltd, we developed HIV gag specific TCRs, a strategy that has recently also shown promise in cancer immunotherapy (Varela et al, in press). The elements of our proposal are: Engineering Elite Control of HIV Infection (Project 1, Jakobsen [Adaptimmune]): This project will clone new HIV-1 specific TCRs, affinity enhance them for improved function, and test specificity and efficacy, and viral evolution/escape (Core C) in vitro and in vivo in the NOG mouse model (Core B). Clinical Trials to Evaluate the Safety and Antiviral Effects of High Affinity Gag Specific CTLs and CCR5- deficient CD4 Cells in HIV/AIDS (Project 2, June [Penn]): This project will test the safety of natural and high affinity HIV specific TCRs in clinical trials, and if safe, evaluate their relative efficiencies via competitive repopulation; in years 4 and 5 we will carry out a second trial combining redirected CTLs with HIV resistant CD4 T cells. Effects on viral evolution (Core C) and HIV specific immunity (Core B) will be studied, along with continued investigation in lentiviral vector integration site analysis (Core A). Zinc Finger Nucleases to Specifically Disrupt HIV Coreceptor Expression (Project 3: Doms [Penn] and Perez [CHOP]). This project will utilize the Sangamo zinc finger nuclease technology to study the effects of CCR5 and CXCR4 knock out in T cells and stem cells, and antiviral effects and viral evolution in vitro and in vivo in the NOG animal model (Cores B and C). The Program is supported by 3 Cores: Core A is the administrative Core (PI, June); Core B is the Novel Immune Assessment and Mouse Core (PI, Riley); and Core C is the Sequencing and Viral Evolution Core (PI, Bushman). In addition, our Program takes advantage of existing School of Medicine (Clinical Cell and Vaccine Production Facility and Human Immunology Core) and CFAR Cores to promote cost sharing and avoid duplication of resources. RELEVANCE: Both the quality of life and economic burdens on the health care system would be improved by obviating the need for patients with HIV infection to take daily antiviral medications. In this proposal we investigate how to genetically engineer a subject's own immune system to teach it how to fight HIV. The scientific principles evaluated in our Program are broadly applicable to other diseases, such as cancer and chronic infection. PROJECT 1: Engineering Elite Control of HIV-1 Infection (Jakobsen, B) PROJECT 1 DESCRIPTION (provided by applicant): While maturing, T cells undergo a massive, largely random rearrangement of their T cell receptor (TCR) genes, resulting in a near unique antigen specificity for each T cell in the body. When a pathogen such as HIV-1 enters the body, T cells recognizing HIV-1 antigens are expanded and a select number of these T cells become overrepresented or immunodominant. However, the ability of these immunodominant HIV-1 specific CDS T cells to control HIV-1 replication varies considerably amongst individuals, and these differences play a major role in determining the rate of disease progression. Individuals able to mount multiple responses targeting HIVGAG have reduced viral loads. Of note, a majority of "elite controllers" express HLA-B alleles associated with potent anti-HIVGAG responses, suggesting that in rare cases effective CDS T cell responses can control HIV-1 infection. However, not all T cell responses targeting HIVGAG are protective and there is no consensus on why this is. Providing insight into why one HIV-1 specific T cell response is more effective than another is a major goal of this project. One way to determine whether one T cell is able to function better than another is to perform population studies in which the presence of a particular T cell response is correlated with viral load. While informative, these studies do not shed light onto why one response is better than another. In vitro studies using HIV-1 specific T cells isolated from individuals indicate that T cells with a higher functional avidity function to control HIV-1 infection better than those with a lower functional avidity, though region of HIV-1 targeted also is important. However, there are several confounding factors including the differentiation state of the T cells and the expression of co-stimulatory and adhesion molecules that preclude direct correlations of functional avidity and TCR affinity. In this application we propose model systems that will permit direct comparison of various HIV-1 specific TCRs and functional avidity through unique proprietary approaches to increase the affinity of natural HlV-specific TCRs and test such responses in vitro and in vivo. RELEVANCE: We predict that these studies will confirm our central hypothesis that non-protective T cell responses such as those that are HLA-A2 restricted can be converted into protective T cell response by engineering higher TCR affinity. Similarly, we aim to investigate whether the introduction of T cells that recognize multiple HIV-1 antigens with high affinity will render HIV-1 sufficiently crippled so that escape does not occur, enabling elite control of HIV-1 infection by autologous T cell transfer.

描述（由申请人提供）：我们的长期目标是建立一种抗HIV的免疫系统。基于先前在HIV感染的免疫发病发生领域的研究，可能需要增强的HLV特异性免疫以及具有增强对HIV感染抗性的CD4细胞群来实现这一目标。在我们正在进行的IPCP拨款（2009年2月到期）中，与Sangamo Biosciences合作，我们使用锌指核酸酶开发了抗HIV的CD4 T细胞，以敲除CCR5共肽，该ccr5 colecector在2009年在临床上进行了测试（Perez等人，印刷中）。为了产生增强的HLV特异性免疫力，与Adaptimmune Ltd合作，我们开发了HIV GAG特异性TCR，该策略最近也显示了癌症免疫疗法的希望（Varela等人，印刷中）。我们提案的要素是：HIV感染的工程精英控制（Project 1，Jakobsen [Adaptimmune]）：该项目将克隆新的HIV-1特定TCR，亲和力增强了它们以提高功能，以及测试特异性和功效，以及病毒进化/逃生（核心C）In Vivo和Invivo In Vivo in Vitro and inog nog nog鼠标模型（Core core Youse b）。评估高亲和力特异性CTL和CCR5缺陷CD4细胞在HIV/AIDS中的临床试验（项目2，6月2日[Penn]）：该项目将测试自然和高亲和力HIV特异性TCR在临床试验中的安全性，如果可以安全地评估其相对效率，请通过有竞争力的重复调查来测试其相对效率的效率；在第4和5年度，我们将进行第二次试验，该试验将重定向的CTL与抗HIV抗HIV CD4 T细胞相结合。将研究对病毒进化（核心C）和HIV特异性免疫（核心B）的影响，并继续研究慢病毒载体整合位点分析（Core A）。锌指核酸酶特异性破坏了HIV共受体表达（项目3：DOMS [PENN]和PEREZ [CHOP]）。该项目将利用Sangamo锌指核酸酶技术来研究CCR5和CXCR4在T细胞和干细胞中敲出的影响，以及NOG动物模型中的体外和体内抗病毒效应和病毒进化（核B和C）。该程序得到了3个核心的支持：核心A是行政核心（PI，6月）；核心B是新型免疫评估和小鼠核心（PI，Riley）；核心C是测序和病毒进化核心（PI，Bushman）。此外，我们的计划利用了现有的医学院（临床细胞和疫苗生产设施以及人类免疫学核心）和CFAR核心来促进成本共享并避免重复资源。 相关性：通过消除艾滋病毒感染的患者每天服用抗病毒药药的需求，可以改善医疗保健系统的生活质量和经济负担。在此提案中，我们调查了如何遗传设计受试者自己的免疫系统来教会它如何与艾滋病毒作斗争。我们计划中评估的科学原理广泛适用于其他疾病，例如癌症和慢性感染。 项目1：HIV-1感染的工程精英控制（Jakobsen，B） 项目1描述（由申请人提供）：在成熟时，T细胞经历了其T细胞受体（TCR）基因的大量随机重排，从而对体内的每个T细胞都具有接近独特的抗原特异性。当像HIV-1这样的病原体进入人体时，识别HIV-1抗原的T细胞会扩大，并且这些T细胞的精选数量变得过分代表或免疫降低。但是，这些免疫主导的HIV-1特异性CD T细胞控制HIV-1复制的能力在个体之间有很大不同，这些差异在确定疾病进展速率方面起着重要作用。能够安装靶向HIVGAG的多个反应的个人减少了病毒载荷。值得注意的是，大多数“精英控制器”表达与有效抗HIVGAG反应相关的HLA-B等位基因，这表明在极少数情况下，有效的CDS T细胞反应可以控制HIV-1感染。但是，并非所有针对HIVGAG的T细胞反应都是保护性的，并且为什么这是什么都没有达成共识。提供有关为什么一个HIV-1特异性T细胞反应比另一个HIV-1特异性T细胞反应更有效的见解是该项目的主要目标。确定一个T细胞是否能够比另一个细胞更好地发挥功能的一种方法是进行人群研究，其中特定T细胞反应的存在与病毒载荷相关。尽管信息丰富，但这些研究并没有阐明为什么一种反应比另一种反应更好。使用从个体分离出的HIV-1特异性T细胞的体外研究表明，功能性较高的T细胞比功能性较低的HIV-1更好地控制HIV-1感染，尽管HIV-1的靶向区域也很重要。但是，有几种混杂因素，包括T细胞的分化状态以及共刺激性和粘附分子的表达，这些分子排除了功能亲和力和TCR亲和力的直接相关性。在此应用中，我们提出了模型系统，该系统将通过独特的专有方法直接比较各种HIV-1特异性TCR和功能亲和力，以增加天然HLV特异性TCR的亲和力，并在体外和体内测试此类反应。 相关性：我们预测，这些研究将证实我们的中心假设，即通过工程提高TCR亲和力，可以将诸如HLA-A2限制的非保护T细胞反应转化为保护性T细胞反应。同样，我们旨在调查引入识别具有高亲和力的多种HIV-1抗原的T细胞是否会使HIV-1充分残废，以免逃脱，从而通过自体T细胞转移对HIV-1感染的精英控制。