Developing Durable, Env-Boosted CAR T Cells for HIV Cure

开发持久的、环境增强的 CAR T 细胞用于治疗 HIV

基本信息

批准号：
10625234
负责人：
Hamideh Parhiz
金额：
$ 89.35万
依托单位：
FRED HUTCHINSON CANCER CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-11-02 至 2027-10-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10625234
关键词：
Address Alleles Animal Model Animals Antibodies Antigens Autologous B lymphoid malignancy B-Lymphocytes Bar Codes Benchmarking Berlin Biological Assay Biology CAR T cell therapy CCR5 gene CD4 Positive T Lymphocytes COVID-19 vaccination COVID-19 vaccine CRISPR/Cas technology CTLA4 gene Cancer Patient Cell Communication Cell Line Cell Therapy Cell physiology Cell surface Cells Chromosomal translocation Clinic Clinical Research Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Data Developing Countries Disease Disease remission Dose Environment Event Experimental Designs Extracellular Domain FDA approved Future Genes Goals Graft-Versus-Tumor Induction HIV HIV therapy HIV-1 Hematologic Neoplasms Hematopoietic Neoplasms Hematopoietic stem cells Homologous Transplantation Immune Individual Infection Interruption K-562 K562 Cells Knowledge Learning Leukemic Cell Liquid substance London Lymphoma cell Macaca mulatta Malignant - descriptor Malignant Neoplasms Messenger RNA Modeling Modification Molecular Nature New York PD-1 blockade Pathway interactions Patients Persons Phase I Clinical Trials Plasma Population Study Procedures Property Public Health Recrudescences Refractory Reporting Research Design Resistance SIV Safety Series Solid Neoplasm Supplementation Supporting Cell T-Lymphocyte Technology Time Vaccines Validation Viral Viral Antigens Viral Envelope Proteins Viral Load result Viral reservoir Virus anti-PD-1 antiretroviral therapy cancer cell cell immortalization chimeric antigen receptor chimeric antigen receptor T cells clinical translation curative treatments env Gene Products exhaustion experimental study gene therapy genotoxicity high risk immune checkpoint immune checkpoint blockade in vivo interest latent HIV reservoir lipid nanoparticle manufacturing process nanoparticle delivery next generation nonhuman primate novel prevent programmed cell death protein 1 programs safety assessment single-cell RNA sequencing stable cell line success trafficking tumor viral rebound

项目摘要

PROJECT SUMMARY/ABSTRACT Modification of autologous T cells with chimeric antigen receptor (CAR) molecules was first proposed nearly 30 years ago as a therapy for people living with HIV. Since then, CAR-T cells have emerged as a potent and highly successful therapy for liquid tumors, while HIV-specific CAR-T cells have only begun to show efficacy in large animal models and clinical trials. Based on our longstanding interest in CAR-T cell therapies for HIV, we posit three primary barriers that limit the curative potential of this approach. First, low levels of HIV-1 antigen at the cell surface (namely Env protein), especially during antiretroviral therapy (ART), render latently infected cells nearly invisible to CAR-T cells and other virus-specific immune effectors. Second, the wealth of knowledge regarding cellular trafficking of the viral Env protein has yet to be thoroughly applied in the context of Env- dependent HIV cure strategies. Third, CAR-T cell persistence and function wane over time, prior to complete clearance of the latent HIV reservoir. Our groundbreaking preliminary data outlines a path to overcome these limitations. We recently reported findings in four rhesus macaques that were infected with an HIV-like virus, suppressed by ART and then infused with virus-specific CAR-T cells containing the CD4 extracellular domain (CD4CAR). To expand these potent antiviral effectors in vivo, animals were next boosted with an irradiated cell line stably expressing HIV-1 Env. Following ART treatment interruption (ATI), viral control was observed in 2 of 4 animals, consistent with robust and Env-dependent expansion of CD4CAR-T cells. The central goal of this proposal is to increase the potency and feasibility of this approach. In AIM 1, we will transition our Env boosting strategy from an immortalized cell line to an FDA-approved mRNA lipid nanoparticle (mRNA-LNP) platform, analogous to the Moderna and Pfizer/BioNTech vaccines for SARS-CoV-2. Env immunogens will be optimized for CD4CAR T cell interactions and developed as Env mRNA-LNP vaccines. In AIM 2, we will use CRISPR- Cas9 gene editing to extend the durability and function of CD4CAR-T cells. We will compare a series of CAR products that carry inactivated immune checkpoint alleles, which we hypothesize will support more durable function and efficiently clear persistent viral reservoirs. In AIM 3, we will benchmark Env mRNA-LNP and immune checkpoint gene editing strategies in our vetted nonhuman primate (NHP) model of HIV gene therapy. These experiments will feature a powerful competitive repopulation study design, providing critical information on basic CAR biology that cannot be gathered in clinical studies. Together, these aims build on what we believe to be the most promising anti-HIV cell and gene therapy approach reported to date. Our unique and highly informative NHP model of HIV persistence and CAR-T cell therapy will fill in critical gaps in knowledge regarding CAR-T cell safety and function in limited antigen environments, and facilitate clinical translation both in developed and developing nations. The lessons we learn from these studies will be applicable not only as a curative therapy for HIV-1, but for a range of diseases such as solid tumors where CAR-T cell therapies must be similarly augmented.

项目概要/摘要用嵌合抗原受体（CAR）分子修饰自体T细胞被首次提出近30 几年前作为一种治疗艾滋病毒感染者的方法。从那时起，CAR-T 细胞已成为一种有效且高度液体肿瘤的成功治疗，而 HIV 特异性 CAR-T 细胞才刚刚开始在大规模治疗中显示出疗效动物模型和临床试验。基于我们对 HIV CAR-T 细胞疗法的长期兴趣，我们认为限制这种方法治疗潜力的三个主要障碍。首先，HIV-1抗原水平低细胞表面（即 Env 蛋白），尤其是在抗逆转录病毒治疗 (ART) 期间，会导致潜伏感染的细胞 CAR-T 细胞和其他病毒特异性免疫效应物几乎看不见。二、丰富的知识关于病毒 Env 蛋白的细胞运输尚未在 Env 的背景下得到彻底应用依赖的艾滋病毒治疗策略。第三，CAR-T细胞的持久性和功能随着时间的推移而减弱，直到完成清除潜伏的艾滋病毒储存库。我们突破性的初步数据概述了克服这些问题的途径限制。我们最近报告了四只感染艾滋病毒样病毒的恒河猴的研究结果，通过 ART 抑制，然后注入含有 CD4 胞外结构域的病毒特异性 CAR-T 细胞（CD4CAR）。为了在体内扩展这些有效的抗病毒效应，接下来用经过辐照的细胞对动物进行增强稳定表达 HIV-1 Env 的细胞系ART 治疗中断 (ATI) 后，2 例患者观察到病毒控制 4 只动物，与 CD4CAR-T 细胞的稳健和 Env 依赖性扩增一致。此次活动的中心目标建议的目的是提高这种方法的效力和可行性。在 AIM 1 中，我们将转变我们的环境提升从永生化细胞系到 FDA 批准的 mRNA 脂质纳米颗粒 (mRNA-LNP) 平台的策略，类似于 Moderna 和 Pfizer/BioNTech 的 SARS-CoV-2 疫苗。 Env免疫原将被优化用于 CD4CAR T 细胞相互作用并开发为 Env mRNA-LNP 疫苗。在 AIM 2 中，我们将使用 CRISPR- Cas9 基因编辑可延长 CD4CAR-T 细胞的耐久性和功能。我们将比较一系列的CAR 携带灭活免疫检查点等位基因的产品，我们假设这些产品将支持更持久的功能并有效清除持久性病毒库。在 AIM 3 中，我们将对 Env mRNA-LNP 和免疫进行基准测试我们经过审查的非人灵长类动物 (NHP) HIV 基因治疗模型中的检查点基因编辑策略。这些实验将以强大的竞争性再群体研究设计为特色，提供有关基本信息的关键信息无法在临床研究中收集的 CAR 生物学信息。这些目标共同建立在我们认为的迄今为止报道的最有前途的抗艾滋病毒细胞和基因治疗方法。我们独特且信息丰富的 HIV持续性和CAR-T细胞疗法的NHP模型将填补有关CAR-T细胞知识的关键空白在有限抗原环境中的安全性和功能，并促进发达国家和地区的临床转化发展中国家。我们从这些研究中吸取的教训不仅适用于治疗 HIV-1，但对于实体瘤等一系列疾病，必须同样增强 CAR-T 细胞疗法。