Use of Genetically Engineered T cells Targeting Tumor Stroma to Treat Lung Cancer

描述(由申请人提供)：免疫治疗的一个特别有希望的领域是通过转移基因编程的患者来源的血淋巴细胞，将嵌合抗原受体基因(CARS)转基因，将T淋巴细胞的效应功能与抗体以非MHC限制性方式识别具有高特异性的预定义表面抗原的能力结合起来。CAR-T细胞治疗的成功取决于靶向一种抗原，这种抗原在肿瘤中高表达，但在正常组织中表达水平很低。到目前为止，只有癌细胞是靶向的，然而，众所周知，除了最小的尺寸之外，癌症的进展取决于包含血管、炎症细胞和成纤维细胞的间质。这一提议的基本假设是破坏 使用CAR-T细胞的间质细胞将改变肿瘤微环境，从而抑制肺癌的肿瘤生长。由于基质细胞是二倍体的，在遗传上稳定，这可以减少免疫逃避的发生率。为了验证这一假设，我们将针对成纤维细胞激活蛋白(FAP)，这是一种基质细胞表面蛋白，在包括肺癌在内的大多数人类上皮癌相关的成纤维细胞上高度表达，但在正常成人组织中水平非常低，除了愈合的伤口和一些慢性炎症条件下。目的1.评价逆转录病毒转导的小鼠FAP-CAR T细胞对肺癌生长的抑制作用(使用三种小鼠肺癌模型)。目的2.探讨逆转录病毒转导的小鼠FAP-CAR T细胞抑制肿瘤生长的机制。目的3.评价FAP-CAR T细胞与化疗(Aim 3A)和肿瘤细胞靶向CAR(Mesothelin-CAR)(Aim 3B)的结合。目的4.评价慢病毒转导的人FAP-CAR T细胞对免疫缺陷小鼠人肺癌生长的抑制作用及其机制。除了获得上述科学问题的答案外，完成这些目标还将提供实施肺癌临床试验所需的临床前数据。本提案没有要求为潜在的FAP-CAR临床试验提供资金，但将通过其他支持获得资金。如果我们的策略被证明是有效的，这个项目可能会有比肺癌更多的意义，因为FAP可以针对许多肿瘤。

DESCRIPTION (provided by applicant): One especially promising area of immunotherapy has been adoptive transfer of genetically programmed, patient-derived blood lymphocytes transfected with chimeric antigen receptor genes (CARs) to combine the effector functions of T lymphocytes with the ability of antibodies to recognize pre-defined surface antigens with high specificity in a non-MHC restricted manner. The success of CAR-T cell therapy depends on targeting an antigen that is highly expressed in tumors, but at minimal levels in normal tissues. To date, only cancer cells have been targeted, however, it is well established that beyond a minimal size, cancer progression is dependent on a stroma that contains blood vessels, inflammatory cells, and fibroblasts. The underlying hypothesis of this proposal is that destruction of stromal cells using CAR-T cells will alter the tumor microenvironment leading to inhibition of tumor growth in lung cancer. Because stromal cells are diploid and genetically stable, this could reduce the incidence of immune evasion. To test this hypothesis, we will target fibroblast activation protein (FAP), a stromal cell-surface protein highly expressed on cancer-associated fibroblasts in most human epithelial cancers, including lung cancers, but present at very low levels on normal adult tissues, except healing wounds and in some chronic inflammatory conditions. We propose the following specific aims: Aim 1. Evaluate the efficacy of retrovirally-transduced murine FAP-CAR T cells to inhibit the growth of lung cancers (using three mouse lung cancer models). Aim 2. Evaluate the mechanisms by which retrovirally-transduced murine FAP-CAR T cells inhibit the growth of tumors. Aim 3. Evaluate the combination of FAP-CAR T cells with chemotherapy (Aim 3A) and with a tumor-cell targeted CAR (mesothelin-CAR) (Aim 3B). Aim 4. Evaluate the efficacy and mechanisms of lentivirally-transduced human FAP-CAR T cells to inhibit the growth of human lung cancers in immunodeficient mice. In addition to obtaining answers to the scientific questions described above, completion of these aims will provide the preclinical data needed to implement a clinical trial for lung cancer. Funding for a potential FAP-CAR clinical trial is not requested in this proposal, but will be obtained through other support. If our strategy proves effective, this project will likely have additional significace beyond lung cancer, since FAP could be targeted in many tumors.