T Cell Receptor - Based Assays for Cancer Diagnosis

描述（由申请人提供）：在过去的十年中，已经鉴定了许多用作T细胞抗原（在MHC蛋白的背景下）的肿瘤相关肽。基于这些发现，以及对T细胞生物学其他方面的最新见解，在开发基于T细胞的癌症疗法方面仍有相当大的努力。正如现在单克隆抗体疗法显而易见的那样，这些T细胞策略的成功将需要诊断测定，以确定患者的癌症是否为抗原阳性（即在其表面上表达肿瘤pep/MHC）。在这项提案中，两名在T细胞受体（TCR）工程（大卫克兰兹）和T细胞定向癌症治疗（菲尔格林伯格）方面具有互补专业知识的研究人员将合作开发基于TCR的癌症诊断检测方法。我们最近已经表明，有可能设计出对pepMHC分子具有特异性的高亲和力TCR。在MHC I类分子的背景下，这些TCR的可溶性形式可以用作新的探针以鉴定和定量肿瘤细胞表面上呈递的肿瘤抗原肽的量。TCR对这些肿瘤抗原之一，肽WT 1的致癌性肾母细胞瘤的肿瘤蛋白，将在本研究中进行探讨。的 该提案的目标是：目标1。开发灵敏的基于T细胞受体的检测方法，用于检测肿瘤细胞上的特异性肽-MHC，使用对pep/MHC复合物SIYR/Kb具有特异性的高亲和力TCR（m67 a）。目标2.针对小鼠WT 1/Dh复合物设计高亲和力TCR，这是一种动物模型，对于开发针对人类WT 1的最佳治疗和诊断至关重要。目标3.设计针对人WT 1/HLA-A2复合物的高亲和力TCR，该抗原复合物已知由各种A2+人类肿瘤内源性表达，包括许多白血病。这些TCR试剂不仅可用于基于检测肿瘤抗原的表达来区分肿瘤细胞与正常细胞，而且还应识别可能最易受免疫干预影响的肿瘤患者。

DESCRIPTION (provided by applicant): In the past decade, many tumor-associated peptides that serve as T cell antigens (in the context of MHC proteins) have been identified. Based on these findings, and recent insights into other aspects of T cell biology, there continues to be considerable effort on the development of T cell-based therapies of cancer. As is now apparent with monoclonal-antibody therapies, success with these T cell strategies will require diagnostic assays that determine whether a patient's cancer is antigen-positive (i.e. expresses the tumor pep/MHC on its surface). In this proposal, two investigators who have complementary expertise in T cell receptor (TCR) engineering (David Kranz) and in T cell-directed therapies of cancer (Phil Greenberg) will collaborate on the development of TCR-based assays for cancer diagnosis. We have recently shown that it is possible to engineer high-affinity TCRs that are specific for pepMHC molecules. Soluble forms of these TCRs can serve as novel probes to identify and quantitate the amount of a tumor antigen peptide presented on the surface of a tumor cell, in the context of an MHC class I molecule. TCRs against one of these tumor antigens, peptide WT1 from the oncogenic Wilm's tumor protein, will be explored in the present study. The goals of this proposal are to: Aim 1. Develop sensitive T cell receptor-based assays for detection of specific peptide-MHC on tumor cells, using an already available high-affinity TCR (m67a) specific for the pep/MHC complex SIYR/Kb. Aim 2. Engineer high-affinity TCRs against mouse WT1/Dh complexes, an animal model that will be critical for the development of optimal therapeutics and diagnostics against WT1 in humans. Aim 3. Engineer high-affinity TCRs against human WT1/HLA-A2 complexes, the antigenic complex known be expressed endogenously by various A2+ human tumors, including many leukemias. These TCR reagents should not only be useful for distinguishing tumor cells from normal cells, based on detecting the expression of a tumor antigen, but they should also identify patients with tumors that might be most susceptible to immunotherapeutic intervention.