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: 8869794
• 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/8869794
• 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; Glycoproteins; Goals; Human; Immunization; Immunology; Immunosuppression; Immunosuppressive Agents; Immunotherapeutic agent; Immunotherapy; Impairment; In Vitro; Inflammation; Inflammatory; Insertional Mutagenesis; Insulin-Dependent Diabetes Mellitus; Knowledge; 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; improved; in vivo; interstitial retinol-binding protein; mouse model; novel; novel strategies; prevent; prototype; public health relevance; 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(申请人提供)：此申请的主要目的是为PI提供研究经验和技能，使其能够在风湿学和眼免疫学之间的基于细胞的免疫疗法开发领域发挥独立研究员的作用。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(纽约大学Juan Lafaille)、风湿病(Steven Abramson，纽约大学)和眼免疫学(Rachel Caspi，NIH-NEI)等领域的国际公认的领导者。申请人的主要职业目标是发展成为一名高素质的内科科学家，具有很强的基础科学技能和将免疫学概念快速转化为临床实践的能力，特别是与开发新的基于细胞的免疫治疗方式有关的免疫学概念。拟议的项目是通过将申请人在白塞氏眼病的临床和实验方面以及细胞免疫学方面已经获得的专业知识纳入并逐步扩展到新的应用、方法和概念，即利用炎症性疾病的人性化动物模型进行体内原理验证和机制研究，从而为这些目标量身定做的。特别是，该项目调查了大规模体外扩增和随后过继转移(重新注射疗法)人类调节性T细胞克隆的可能性，以控制自身免疫性葡萄膜炎的致病效应T细胞(TEF)反应。它测试了主要假设，即人类的CD4+/CD25hi/CD127lo/Foxp3+/Helios+表型[胸腺，或自然(N)Treg]的Treg在体外大规模扩增后在体外和体内保持功能稳定，并有效地控制一般和/或靶(葡萄膜炎)抗原相关效应T细胞的反应。申请人已经证明了在维持体外抑制功能的情况下克隆和大规模克隆nTreg是可能的，他现在建议在特定目标1中巩固这一方法，并将其有效性与多克隆现有方法进行比较。特异性目标2验证了nTreg可以在已知其抗原特异性的情况下被克隆的假设。他将使用他之前在树突状细胞介导的T细胞启动和扩增方面获得的指导工作中获得的技能，并将葡萄膜炎抗原特异的免疫优势和致病性方面作为主要的教育和科学目标。具体目标3)以双尾方法在体内验证和测试单克隆化扩增的Treg：Subaim a)将测试具有未选定抗原特异性的克隆的一般免疫抑制功能，在人源化的移植物抗宿主病(GVHD)小鼠模型中，以确定和量化体内抑制，以及亚目标b)将建立人源化的实验性自身免疫性葡萄膜炎(EAU)小鼠模型，用于具有葡萄膜炎抗原特异性的nTreg克隆的临床前测试。从拟议的研究中获得的知识有可能显著改进目前的Treg扩增方法和采用的转移策略，这些方法目前主要依赖于多克隆技术，并受到潜在有害的TREF的有害共扩展、不稳定的抑制功能和低扩张率的限制。抗原特异性克隆方法还具有在病理部位靶向抑制TJeff反应的潜力，而不会出现明显的全身免疫抑制，并消除了插入突变和双T细胞受体(TCR)表达的风险，目前流行的策略是通过将慢病毒TCR转导到多克隆扩增的Treg中来赋予抗原特异性。除了针对自身免疫性葡萄膜炎的一种新的靶向治疗方式的潜力外，这项研究的结果可能对其他形式的自身免疫性疾病具有高度的推广性。