Advanced generation infection-proof anti-HIV CAR-T with YY1 RNAi to block T cell exhaustion in NHP model

新一代防感染抗 HIV CAR-T 具有 YY1 RNAi，可阻止 NHP 模型中的 T 细胞耗竭

基本信息

批准号：
10689338
负责人：
Richard P Junghans
金额：
$ 100万
依托单位：
IT BIO, LLC
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-17 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10689338
关键词：
Address Affect Antibodies Antigens Antisense Technology Area B-Lymphocytes Boston CCR5 gene CD28 gene CD4 Antigens CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Cell Therapy Cell physiology Cells Chronic Clinical Clinical Trials Collaborations Contract Services Cytotoxic T-Lymphocytes Down-Regulation Engineering Ensure Environment Failure Fee-for-Service Plans Generations Goals HIV HIV Envelope Protein gp120 HIV Infections HIV drug resistance HIV therapy HIV-1 Human Immune Immunity Immunologic Surveillance In Vitro Infection Inflammatory Infusion procedures Interferon Type II Interleukin-2 Intervention Investigation Knock-out Laboratories Ligands Malignant Neoplasms Mediating Methods Modeling Molecular Monkeys PD-1 blockade Patients Phase Preclinical Testing Prevention Primates Process Production RNA Interference Research Resistance Rest Side Signal Transduction Small Business Innovation Research Grant Structure Surveys T-Lymphocyte Tars Techniques Technology Testing Text Therapeutic Therapeutic Agents Universities Up-Regulation Validation Viral Viral Proteins Virus Latency Virus Replication Work YY1 Transcription Factor base cancer cell checkpoint receptors chimeric antigen receptor chimeric antigen receptor T cells chronic infection cytokine cytotoxic design effector T cell env Gene Products exhaust exhaustion experimental study improved in vitro Assay in vivo in vivo evaluation knock-down neoplastic cell nonhuman primate novel pre-clinical preservation programmed cell death protein 1 rational design receptor receptor expression response small hairpin RNA success vector

项目摘要

This FastTrack Phase 1/2 application aims to create a new IND-ready CAR-T platform for the treatment of HIV, developed in the laboratory and tested in non-human primates. Molecular engineering techniques have been applied to create chimeric antigen receptors (CAR) expressed in T cells to target HIV-infected cells. CD4-based CARs are designed to achieve immune eradication of HIV1 infections through recognizing gp120 envelope protein on infected cells. However, prior clinical trials did not meet with success, which we propose to address with the plan of this research. One of the features predicted to affect efficiency of CD4 CAR is T cell exhaustion, characterized by high PD1 expression of HIV-specific CD8 and CD4 T cells. Our laboratory recently defined transcription factor YY1 to be master regulator of T cell exhaustion, mediating upregulation of checkpoint receptors (CR) and downregulation of Type I cytokines with accompanying cytotoxic failure. We confirmed that YY1 is increased in parallel with PD1 in CD4 T cells in chronic HIV infection. Knockdown of YY1 restored cytokine IL2 production in preclinical testing and reduced CR expression, including exhaustion marker PD1, where blocked PD1 and restored IL2 correlated with recovered T cell cytotoxic potency. Another drawback of prior CD4- based CAR-T is that CD8 T cells expressing the CD4 CAR receptor are now readily infected and eliminated by HIV that could also have hampered success of prior human trials. Lastly, prior tests involved 1st generation (gen) CARs of limited signaling potential that are now improved with addition of costimulation that may be yet further improved. Our overall goal is to create a more effective CAR-T for the control of HIV. Our Aims for this proposal include (1) creating anti-HIV CAR-T cells that will resist T cell exhaustion with incorporation of YY1 shRNA for sustained anti- HIV potency. Further, we will (2) render the CD4 CAR-T infection-proof with RNAi intervention to block infection and virus replication in the CAR-T. Finally, we will (3) conduct a complementary effort to generate new 3rd gen 3-signal CD4 CARs (CAR3) that incorporate additional costimulatory molecules to improve potency and reactivation potential. Building on our considerable preliminary work, we will quickly finish the Phase 1 component to complete molecular engineering efforts in months 1-6, moving directly to Phase 2 in vitro and then non- human primate testing. This plan is a collaboration between IT Bio, LLC (Boston) and the lab of Dr Steven Braun (Tulane). The Tulane National Primate Research Center (TNPRC) will conduct the NHP experiments under the direction of IT Bio, LLC through a fee-for-service contract. With these three efforts – suppressing T cell exhaustion, rendering CAR-T infection-proof and increasing T cell potency and reactivation potential – we hope to obtain a novel, IND-ready cellular therapeutic agent that will provide a sustained control of HIV to parallel recent successes in CAR- T treatment of B cell cancers.

这个FastTrack阶段1/2应用程序旨在为该新的Ind-Ready Car-T平台创建在实验室开发并在非人类隐私中进行了测试的艾滋病毒的治疗。分子已经应用了工程技术来创建表达的嵌合抗原受体（CAR）在T细胞中靶向HIV感染细胞。基于CD4的汽车旨在实现免疫通过识别感染细胞上的GP120包膜蛋白来消除HIV1感染。但是，先前的临床试验没有获得成功，我们建议解决该计划这项研究。预计会影响CD4 CAR效率的功能之一是T细胞耗尽，以HIV特异性CD8和CD4 T细胞的高PD1表达为特征。我们的实验室最近定义的转录因子yy1是T细胞耗尽的主要调节剂，介导检查点受体（CR）的上调和I型细胞因子的下调参与细胞毒性衰竭。我们确认YY1与CD4中的PD1并行增加慢性HIV感染中的T细胞。 yy1的敲低恢复了临床前的细胞因子IL2产生测试和降低CR表达，包括耗尽标记PD1，其中PD1和恢复的IL2与回收的T细胞细胞毒性效力相关。先前CD4-的另一个缺点基于CAR-T是表达表达CD4 CAR受体的CD8 T细胞现在很容易感染，并且被HIV淘汰，这也可能阻碍了先前的人类试验的成功。最后，先前的测试涉及有限信号电势的第一代汽车，现在可以通过添加的共刺激可能会进一步改善。我们的总体目标是创建更多有效控制艾滋病毒的CAR-T。我们对该建议的目标包括（1）创建反HIV CAR-T细胞将通过掺入YY1 shRNA来抵抗T细胞耗尽艾滋病毒效力。此外，我们（2）将CD4 CAR-T型通过RNAi干预介绍阻止CAR-T中的感染和病毒复制。最后，我们将（3）进行完整的努力生成新的3代信号CD4汽车（CAR3），该汽车包含其他共刺激分子以提高效力和重新激活潜力。建立在我们的基础上相当大的初步工作，我们将迅速完成1阶段组件以完成在1-6个月的分子工程工作，直接在体外转移到第2阶段，然后非 - 人类灵长类动物测试。该计划是IT Bio，LLC（波士顿）与实验室的合作史蒂文·布劳恩（Steven Braun）博士（图兰）。图兰国家灵长类动物研究中心（TNPRC）将进行 NHP在IT Bio，LLC下通过费用服务合同进行的实验。和这三个努力 - 抑制T细胞耗尽，使CAR-T防护感染和提高T细胞的效力和重新激活潜力 - 我们希望获得一种新颖的，准备的细胞治疗剂将持续控制艾滋病毒，从而相似 t治疗B细胞癌。