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Engineering T Cell Receptors for Adoptive Cell Therapies

工程 T 细胞受体用于过继细胞疗法

基本信息

批准号：
8631350
负责人：
David M. Kranz
金额：
$ 32.05万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8631350
关键词：
Affinity Antibodies Antigen Receptors Antigens Biological Models CD19 gene CD28 gene CD4 Positive T Lymphocytes CD8B1 gene Cell Fraction Cell Therapy Cell Transplants Cell surface Collaborations Complex ERBB2 gene Effectiveness Engineering Evolution Exhibits HLA-A2 Antigen Health Human Human Engineering Immunoglobulin Fragments In Vitro Indium Libraries Ligands Link MS4A1 gene Malignant Neoplasms Mediating Memory Modeling Mus Normal tissue morphology PTEN gene Peptide/MHC Complex Peptides Peripheral Process Property Receptor Signaling Reporting Risk Signal Transduction Signaling Protein Specificity Structure System T cell therapy T-Cell Receptor T-Lymphocyte Techniques Time Transgenic Mice Transplantation Tumor Antigens Virus WT1 gene Yeasts base cancer cell cancer therapy chimeric antigen receptor cross reactivity design improved in vivo interest melanoma novel novel strategies receptor research study response scaffold tumor

项目摘要

DESCRIPTION (provided by applicant): Adoptive T cell therapy can be an effective and highly specific approach to the treatment of cancer. Introduction of receptors that recognize cancer antigens provides sufficient numbers of T cells with the appropriate specificity to destroy large, established tumors. The receptors used to date include either αβ T cell receptors (TCRs) against pepMHC or antibody (scFv)-directed chimeric antigen receptors (CARs) against conventional cell surface cancer antigens. The purpose of this proposal is to improve upon current therapies and to develop a robust strategy that combines the advantages of each of these targeting approaches. The strategy involves a single-chain TCR (Vα-linker-Vβ, called scTv) fused to signaling domains of CD28 (or 4-1BB) and CD3ζ. When endowed with a high-affinity scTv, this novel signaling receptor, which we call TCR-SCS (TCR-Single-Chain Signaling fusions), redirects activity of both CD4 and CD8 T cells. The TCR-SCS can be used without some of the risks associated with conventional αβ TCRs. For example, TCR-SCS receptors limit self-peptide, off-target reactivities and they avoid mis-pairing with endogenous TCR chains. The reduced levels of off-target reactivities arise from the inability of the single-chain signaling protein to synergize with CD8, a process that enhances sensitivity in CD8 T cells but also increases the level of stimulation by other pepMHC. To enable the rapid use of these TCR-SCS receptors in human therapies, we will also use structure-guided design to engineer high-affinity TCRs against specific pep/HLA-A2 antigens. Our central hypotheses are that TCR-SCS (TCR-Single-Chain Signaling fusions) can be rapidly engineered against many different pepMHC target antigens, and that the TCR-SCS will mediate effective CD4 and CD8 T cell activity without off-target cross-reactivity. Accordingly, the specific aims are: Aim 1. To isolate human T cell receptors against diverse peptide/HLA-A2 antigens using a single TCR scaffold. The approach will involve a combination of structure-based design, taking advantage of computational analyses for library construction and advanced techniques of yeast display for the rapid isolation and evolution of the TCRs. Aim 2. To determine if a TCR-SCS (TCR-Single-Chain Signaling fusion) influences T cell persistence and function in mice. Because the TCR-SCS format is a completely novel approach, we will use the model system involving the peptide SIY, and the high-affinity m33 TCR-SCS against SIY/Kb, to further examine in vivo properties of transduced CD4 and CD8 T cells (collaboration with Dr. Hans Schreiber). Tumor models will include an inducible B-Raf/PTEN-/- melanoma that expresses SIY/Kb (collaboration with Dr. Tom Gajewski). Aim 3. To use TCR-SCS (TCR-Single-Chain Signaling fusions) with high-affinity, human TCRs against WT1 to target tumors. Established, transplanted WT1/HLA-A2 human tumors will be used to examine effectiveness of CD4 and CD8 T cells, transduced with WT1-specific TCR-SCS receptors (collaboration with Dr. Philip Greenberg). Preliminary studies with TCRs isolated in Aim 1 will also extend results to other human tumor antigens.

描述（由申请人提供）：过继性 T 细胞疗法可能是一种有效且高度特异性的癌症治疗方法。引入识别癌症抗原的受体可提供足够数量的具有适当特异性的 T 细胞，以摧毁已形成的大型肿瘤。迄今为止使用的受体包括针对 pepMHC 的 αβ T 细胞受体 (TCR) 或针对传统细胞表面癌抗原的抗体 (scFv) 定向嵌合抗原受体 (CAR)。该提案的目的是改进当前的治疗方法，并制定结合每种靶向方法的优点的稳健策略。该策略涉及将单链 TCR（Vα-连接子-Vβ，称为 scTv）与 CD28（或 4-1BB）和 CD3ze 的信号域融合。当赋予高亲和力 scTv 时，这种新型信号受体（我们称之为 TCR-SCS（TCR-单链信号融合））会重定向 CD4 和 CD8 T 细胞的活性。 TCR-SCS 的使用不会产生与传统 αβ TCR 相关的一些风险。例如，TCR-SCS 受体限制自肽、脱靶反应，并避免与内源 TCR 链错误配对。脱靶反应水平降低是由于单链信号蛋白无法与 CD8 协同作用，这一过程增强了 CD8 T 细胞的敏感性，但也增加了其他 pepMHC 的刺激水平。为了能够在人类治疗中快速使用这些 TCR-SCS 受体，我们还将使用结构引导设计来设计针对特定 pep/HLA-A2 抗原的高亲和力 TCR。我们的中心假设是，TCR-SCS（TCR-单链信号融合）可以针对许多不同的 pepMHC 靶抗原进行快速工程设计，并且 TCR-SCS 将介导有效的 CD4 和 CD8 T 细胞活性，而不会出现脱靶交叉反应。因此，具体目标是：目标 1. 使用单一 TCR 支架分离针对不同肽/HLA-A2 抗原的人 T 细胞受体。该方法将结合基于结构的设计，利用文库构建的计算分析和酵母展示的先进技术来快速分离和进化 TCR。目标 2. 确定 TCR-SCS（TCR-单链信号融合）是否影响小鼠中 T 细胞的持久性和功能。由于 TCR-SCS 格式是一种全新的方法，因此我们将使用涉及肽 SIY 的模型系统以及针对 SIY/Kb 的高亲和力 m33 TCR-SCS，以进一步检查转导的 CD4 和 CD8 T 细胞的体内特性（与 Hans Schreiber 博士合作）。肿瘤模型将包括表达 SIY/Kb 的诱导型 B-Raf/PTEN-/- 黑色素瘤（与 Tom Gajewski 博士合作）。目标 3. 使用针对 WT1 的高亲和力人类 TCR 的 TCR-SCS（TCR-单链信号融合）来靶向肿瘤。已建立的移植 WT1/HLA-A2 人类肿瘤将用于检查用 WT1 特异性 TCR-SCS 受体转导的 CD4 和 CD8 T 细胞的有效性（与 Philip Greenberg 博士合作）。对目标 1 中分离的 TCR 进行的初步研究还将结果扩展到其他人类肿瘤抗原。