A comparative analysis of human and canine iNKT cells for ACT

人和犬 iNKT 细胞 ACT 的比较分析

基本信息

批准号：
10287095
负责人：
Qi Long
金额：
$ 23.85万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10287095
关键词：
Address Adoptive Cell Transfers Adoptive Transfer Agonist Allogenic Antibodies Autologous Award Bioinformatics Biological Biology CD94 Antigen Cancer Patient Canis familiaris Cell Therapy Cell physiology Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Client Clinic Clinical Clinical Trials Cytotoxic T-Lymphocytes Data Development Epitopes Evaluation Exhibits Frequencies Future Future Generations Galactosylceramides Gene Silencing Generations Genetic Transcription Glycolipids Goals Heterogeneity Human Immune Immunocompetent Immunologics Immunologist Instruction Investigation Investigational Therapies Libraries Lymphoma MHC Class I Genes Malignant Neoplasms Membrane Proteins Monoclonal Antibodies Mus Natural Killer Cells Oncology Organ Organ Transplantation Outcome Parents Patients Performance Phage Display Phenotype Population Population Heterogeneity Pre-Clinical Model Prevention trial Procedures Property Protocols documentation Psychological Transfer Reagent Solid Sorting - Cell Movement Surface T-Lymphocyte T-Lymphocyte Subsets TRAF4 gene Testing Therapeutic Therapeutic Effect Therapy trial Translations Transplantation Tropism Tumor Immunity Tumor-associated macrophages Work base cancer cell cancer immunotherapy clinical effect comparative cytotoxic experimental study genome editing immunoregulation immunotherapy clinical trials improved in vivo in vivo evaluation indexing interest lymphoid irradiation member mouse model novel phenotypic biomarker receptor response single-cell RNA sequencing transcriptome transcriptome sequencing transcriptomics tumor tumor microenvironment

项目摘要

PROJECT SUMMARY (See instructions): The holy grail of adoptive cell therapies (ACT) is the generation of fit, ready-to-use, CART products from healthy donors that could be safely and effectively transferred across allogeneic barriers to improve the availability and efficacy of CAR-therapies. Invariant Natural Killer T cells (iNKT) are a heterogenous population of `non-conventional' T cells expressing an invariant TCRα which recognizes the non-polymorphic, glycolipid- presenting MHC molecule CD1d. As such, unedited allogeneic iNKT cells (allo-iNKT) can be adoptively transferred without causing GVHD. Further, iNKT cells suppress GVHD caused by conventional T cells and inhibit rejection of allogeneic cells promoting lifelong tolerance of solid organ grafts. In addition, iNKT cells have natural tumor tropism, kill CD1d+ tumor associated macrophages and activate effector immune cells in the tumor. Given these properties, we hypothesize that allo-iNKT could be safely and effectively used to promote allo-CART persistence and enhance anti-tumor immunity. The complexity of iNKT function may be attributed to distinct iNKT subsets that exhibit cytotoxic versus regulatory properties. Understanding subset heterogeneity would enable the most “desirable” iNKT subsets to be selected for evaluation of combination ACT. Unlike mice, canine iNKT cells share similar phenotypic and biological features to human iNKT cells, suggesting that pet dogs with spontaneous cancer may represent the ideal pre-clinical model to investigate combination allo-ACT using iNKT cells. Here, in alignment with the aims of PRECINCT to support canine immunotherapy clinical trials and generate immunological reagents and with the interest of IOTN to generate an off-the-shelf CART cell platform, we will perform a comparative approach to investigate actionable iNKT cell heterogeneity through an integrated, single cell RNAseq/CITEseq approach in human iNKT and employ transfer learning to impute phenotypic markers of canine iNKT subsets from scRNA seq data. Furthermore, to expand the immunoreagent toolbox for canine NK studies, a comprehensive scFv phage display library will be used in simple panning experiments to generate and validate much needed canine- specific antibodies against NK activation and inhibitory receptors that will be of value in understanding the mechanisms governing canine NK activation and inhibition. Together, these studies will lay the necessary groundwork for future generation and in vivo testing of optimal iNKT cell products for combination allo- iNKT/CART therapies in immune competent canine cancer patients, with the goal to accelerate novel allo-ACT strategies into the human clinic. RELEVANCE (See instructions): The natural properties of iNKT cells lend themselves to safe allogeneic adoptive transfer and suppression of alloreactive responses that would otherwise eliminate allo-CART products. Here, we will provide the necessary comparative evaluation of human and canine iNKT cells that will enable future in vivo testing of optimal, translationally relevant iNKT products for combination allo-iNKT/CART therapies in immune competent canine cancer patients, aiming to accelerate novel allo-ACT strategies into the human clinic

项目摘要（参见说明）：过继细胞疗法 (ACT) 的目标是生成适合的、即用型 CART 产品健康的捐赠者可以安全有效地跨越同种异体障碍进行转移，以改善 CAR 疗法的可用性和疗效。表达恒定 TCRα 的“非常规”T 细胞，该 TCRα 可识别非多态性糖脂呈递 MHC 分子 CD1d 因此，未经编辑的同种异体 iNKT 细胞 (allo-iNKT) 可以被过继地使用。此外，iNKT 细胞可抑制传统 T 细胞引起的 GVHD，并且不会引起 GVHD。抑制同种异体细胞的排斥反应，促进实体器官移植物的终生耐受。此外，iNKT 细胞还具有以下作用：天然肿瘤趋向性，杀死CD1d+肿瘤相关巨噬细胞并激活肿瘤中的效应免疫细胞。鉴于这些特性，我们追求allo-iNKT可以安全有效地用于推广allo-CART 持久性并增强抗肿瘤免疫力。 iNKT 功能的复杂性可能归因于不同的 iNKT 子集，它们表现出细胞毒性与了解子集异质性将使最“理想的”iNKT 子集能够与小鼠不同，犬 iNKT 细胞具有相似的表型和特征。人类 iNKT 细胞的生物学特征表明，患有自发性癌症的宠物狗可能代表了使用 iNKT 细胞研究联合 allo-ACT 的理想临床前模型。 PRECINCT 支持犬免疫治疗临床试验并生产免疫试剂由于 IOTN 有兴趣生成现成的 CART 单元平台，我们将采用比较方法来通过集成的单细胞 RNAseq/CITEseq 方法研究可操作的 iNKT 细胞异质性人类 iNKT 并利用迁移学习从 scRNA 推断犬 iNKT 子集的表型标记此外，为了扩展犬 NK 研究的免疫试剂工具箱，我们还提供了综合性的 scFv。噬菌体展示库将用于简单的淘选实验，以生成和验证急需的犬类针对 NK 激活和抑制受体的特异性抗体，对于理解 NK 激活和抑制受体具有重要价值这些研究将为犬类 NK 激活和抑制的机制奠定基础。为组合异体 iNKT 细胞产品的未来最佳生成和体内测试奠定基础对免疫能力强的犬癌症患者进行 iNKT/CART 疗法，目标是加速新型 allo-ACT 的开发策略进入人类临床。相关性（参见说明）： iNKT 细胞的天然特性有助于安全的同种异体过继转移和抑制的同种反应反应，这将消除其他同种异体 CART 产品。对人类和犬 iNKT 细胞进行必要的比较评估，这将使未来的体内测试成为可能用于免疫能力强的同种异体 iNKT/CART 联合疗法的最佳、翻译相关的 iNKT 产品犬癌症患者，旨在加速新型 allo-ACT 策略进入人类临床