Harnessing monoclonal Treg for the treatment of autoimmune uveitis

利用单克隆 Treg 治疗自身免疫性葡萄膜炎

• 批准号: 9045643
• 负责人: Johannes Nowatzky
• 金额: $ 18.14万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9045643
• 关键词: Adoptive Transfer; Adverse drug effect; Adverse effects; Affect; American; Animal Model; Antigens; Arrestins; Autoimmune Diseases; Autoimmunity; Autologous; Basic Science; Biology; Cells; Cellular Immunology; Clinical; Cloning; Dendritic Cells; Development; Disease; Epitopes; Eragrostis; Eye; Eye diseases; Gene Transfer; Goals; Health; Human; Immunization; Immunology; Immunosuppression; Immunosuppressive Agents; Immunotherapeutic agent; Immunotherapy; Impairment; In Vitro; Inflammation; Inflammatory; Insertional Mutagenesis; Insulin-Dependent Diabetes Mellitus; Knowledge; LRRC32 gene; Maintenance; Mediating; Mentors; Methodology; Methods; Modality; Modeling; Multiple Sclerosis; Mus; Mutagenesis; Pathogenicity; Pathology; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Physicians; Preclinical Testing; Preparation; Proteins; Qualifying; Reapplication; Regulatory T-Lymphocyte; Research; Research Personnel; Rheumatoid Arthritis; Rheumatology; Risk; Scientist; Self Tolerance; Site; Specificity; Staging; System; T cell response; T-Cell Receptor; T-Cell Receptor Genes; T-Lymphocyte; Tail; Techniques; Testing; Therapeutic; Translations; United States; United States National Institutes of Health; Up-Regulation; Ursidae Family; Uveitis; Validation; Viral; Vision; Wit; Work; autoimmune uveitis; base; career; cell type; clinical practice; cohort; effective therapy; experience; graft vs host disease; humanized mouse; improved; in vivo; interstitial retinol-binding protein; mouse model; novel; novel strategies; prevent; prototype; receptor expression; reconstitution; response; skills; targeted treatment

 DESCRIPTION (provided by applicant): The main objective of this application is to provide the PI with research experience and skills which will enable him to function as an independent investigator in the field of cell-based immunotherapy development at the interface between rheumatology and ocular immunology. To this end, the candidate has assembled a mentoring team which includes internationally-recognized leaders in the fields of regulatory T cell (Treg) biology (Juan Lafaille, NYU), rheumatology (Steven Abramson, NYU), and ocular immunology (Rachel Caspi, NIH-NEI). The overarching career goal of the applicant is to develop into a highly qualified physician-scientist with strong basic science skills AND the capacity for rapid translation of immunology concepts to aspects of clinical practice, especially as pertains to the development of new cell-based immunotherapeutic modalities. The proposed project is tailored to these goals through the incorporation and step-wise extension of expertise already acquired by the applicant in clinical and experimental aspects of Behcet's eye disease, and cellular immunology, to new applications, methods and concepts, i.e., experimentation with humanized animal models of inflammatory disease for in vivo proof-of-principle and mechanistic studies. In particular, the project investigates the possibility of massive ex vivo expansion and subsequent adoptive transfer (reinjection therapy) of human regulatory T cell clones for the control of pathogenic effector T cell (Teff) responses in autoimmune uveitis. It tests the main hypothesis that human Treg of the CD4+/CD25hi/CD127lo/Foxp3+/Helios+ phenotype [thymic, or natural (n)Treg] maintain functional stability in vitro and in vivo following massive monoclonal ex vivo expansion, and effectively control general and/or target (uveitis) antigen-related effector T cell responses. The applicant has shown that cloning and massive monoclonal expansion of nTreg under maintenance of in vitro suppressor function is possible, and he now proposes to solidify this approach in Specific Aim 1 and contrast its efficacy with polyclonal, existing approaches. Specific Aim 2 tests the hypothesis that nTreg can be cloned with knowledge of their antigen-specificity. He will use the skills acquired in his previous mentored work in dendritic cell-mediated T cell priming and expansion and incorporate uveitis antigen-specific aspects of immunodominance and pathogenicity as the main educational and scientific objective. Specific Aim 3 focuses on in vivo validation and testing of monoclonally expanded Treg in a 2-tailed approach: Subaim a) will test general immunosuppressive function of clones with unselected antigen-specificity the PI has already generated, in a humanized mouse model of graft-versus-host-disease (GVHD) to determine and quantify in vivo suppression, and subaim b) will establish a humanized experimental autoimmune uveitis (EAU) mouse model for preclinical testing of nTreg clones with uveitis antigen specificity. The knowledge to be gained from the proposed studies bears the potential to significantly improve current approaches of Treg expansion and adoptive transfer strategies, which at present rely mostly on polyclonal techniques and are limited by the unwanted co-expansion of potentially harmful Teff, unstable suppressor function and low expansion rates. The antigen-specific cloning approach further bears the potential of targeted suppression of Teff responses at the site of pathology without significant systemic immunosuppression and eliminates the risks of insertional mutagenesis and dual T cell receptor (TCR) expression inherent in the currently prevailing strategies that confer antigen specificity through lentiviral TCR transduction into polyclonally expanded Treg. Apart from the potential of a new modality of targeted treatment for autoimmune uveitis, results of this research are likely to have a high degree of generalizability to other forms of autoimmune diseases.

 描述（由申请人提供）：本申请的主要目的是为PI提供研究经验和技能，使其能够在风湿病学和眼免疫学之间的界面上作为基于细胞的免疫疗法开发领域的独立研究者。为此，候选人组建了一个指导团队，其中包括调节性T细胞（Treg）生物学（Juan Lafaille，纽约大学），风湿病学（Steven Abramson，纽约大学）和眼免疫学（Rachel Caspi，NIH-NEI）领域的国际公认领导者。申请人的总体职业目标是发展成为一名高素质的医生-科学家，具有较强的基本科学技能，并能够将免疫学概念快速转化为临床实践的各个方面，特别是与开发新的基于细胞的免疫学模式有关。拟议项目是针对这些目标，通过合并和逐步扩展申请人在白塞氏眼病的临床和实验方面已经获得的专业知识，以及细胞免疫学，以新的应用，方法和概念，即，用炎性疾病的人源化动物模型进行实验，用于体内原理验证和机理研究。特别是，该项目研究了人类调节性T细胞克隆的大规模离体扩增和随后过继转移（再注射治疗）的可能性，以控制自身免疫性葡萄膜炎中的致病性效应T细胞（Teff）反应。它测试了主要假设，即CD 4 +/CD 25 hi/CD 127 lo/Foxp 3 +/Helios+表型的人Treg [胸腺或天然（n）Treg]在大规模单克隆离体扩增后在体外和体内保持功能稳定性，并有效控制一般和/或靶（葡萄膜炎）抗原相关的效应T细胞应答。申请人已经表明，在维持体外抑制功能的情况下，nTreg的克隆和大规模单克隆扩增是可能的，并且他现在提出在特异性目的1中巩固这种方法，并将其功效与多克隆现有方法进行对比。特异性目的2检验了nTreg可以在了解其抗原特异性的情况下克隆的假设。他将使用他以前在树突状细胞介导的T细胞引发和扩增的指导工作中获得的技能，并将免疫优势和致病性的葡萄膜炎抗原特异性方面作为主要的教育和科学目标。具体目标3侧重于在双尾方法中对单克隆扩增的Treg进行体内验证和测试：Subaim a）将在移植物抗宿主病（GVHD）的人源化小鼠模型中测试PI已经产生的具有IgM抗原特异性的克隆的一般免疫抑制功能，以确定和定量体内抑制，子目标B）将建立人源化实验性自身免疫性葡萄膜炎（EAU）小鼠模型，用于临床前测试具有葡萄膜炎抗原特异性的nTreg克隆。从所提出的研究中获得的知识有可能显著改善Treg扩增和过继转移策略的当前方法，这些方法目前主要依赖于多克隆技术，并且受到潜在有害的Teff的不需要的共扩增、不稳定的抑制功能和低扩增速率的限制。抗原特异性克隆方法还具有在病理部位靶向抑制Teff应答而不产生显著的全身性免疫抑制的潜力，并且消除了目前流行的策略中固有的插入诱变和双重T细胞受体（TCR）表达的风险，所述策略通过慢病毒TCR转导到多克隆扩增的Treg中来赋予抗原特异性。除了自身免疫性葡萄膜炎的靶向治疗的新模式的潜力，这项研究的结果很可能具有高度的普遍性，以其他形式的自身免疫性疾病。