T cell plasticity, fusion proteins and CAR T cell-based immunotherapy of head and neck cancer

T细胞可塑性、融合蛋白和基于CAR T细胞的头颈癌免疫疗法

基本信息

批准号：
9982679
负责人：
SOLDANO FERRONE
金额：
$ 38.7万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9982679
关键词：
Address Adoptive Immunotherapy Adrenal Glands Affect Anatomy Antigens Apoptosis Apoptotic B-Cell Leukemia Biological Models CD276 gene Cell Hypoxia Cell Line Cell Survival Cell physiology Cells Cellular immunotherapy Characteristics Chimeric Proteins Chondroitin Sulfate A Chondroitin Sulfate Proteoglycan Combination immunotherapy Cytolysis Cytotoxic T-Lymphocytes Disease Drug resistance Epitopes Fibroblasts Functional disorder Generations Genes Genetic Transcription Growth Head and Neck Cancer Head and Neck Neoplasms Head and Neck Squamous Cell Carcinoma Heterogeneity Homologous Gene Human Hypoxia I-antigen Immunodeficient Mouse Immunotherapy Impairment In Vitro Incubated Interleukin-15 Interleukin-2 Link Longevity Lytic Malignant Neoplasms Mediating Membrane Monoclonal Antibodies Mouse Strains Mus Non-Malignant Normal tissue morphology Operative Surgical Procedures Oxygen PD-1/PD-L1 Parotid Gland Pathway interactions Patients Pericytes Phase I Clinical Trials Play Predisposition Publishing Recurrence Reporting Resistance Rest Role SHH gene Solid Neoplasm Squamous Differentiation T-Lymphocyte Testing Treatment Efficacy Tumor Antigens Tumor Necrosis Factor Ligand Superfamily Member 6 Up-Regulation Xenograft procedure aldehyde dehydrogenases angiogenesis anti-PD-L1 cancer stem cell cellular transduction chimeric antigen receptor chimeric antigen receptor T cells clinically relevant clinically significant combinatorial cytokine cytotoxic design effective therapy engineered T cells exhaustion experimental study improved in vivo mouse model neoplastic cell novel preservation programmed cell death ligand 1 programmed cell death protein 1 self-renewal side effect small molecule stemness theories therapeutic evaluation treatment strategy tumor tumor hypoxia tumor microenvironment tumor specificity tumorigenic

项目摘要

ABSTRACT The limited efficacy of the available therapy for squamous cell carcinoma of the head and neck (SCCHN) has prompted us to design a novel effective combinatorial immunotherapy for this disease. In this strategy, T cells engineered with a tumor antigen (TA)-specific chimeric antigen receptor (CAR) are used as the effector mechanism, since this approach allows specificity of tumor recognition and self amplification due to T lymphocyte self renewal capacity. We have selected chondroitin sulfate proteoglycan 4 (CSPG4) as the target, since i) CSPG4 is highly expressed in about 60% of SCCHN tumors with limited heterogeneity within each tumor; ii) CSPG4 is expressed on both differentiated SCCHN cells and SCCHN cells with high aldehyde dehydrogenase activity. The latter cells referred to as ALDHbright cells, display the characteristics of cancer initiating cells (CICs), since they are drug-resistant, express stemness genes and are tumorigenic in immunodeficient mice. Therefore, CSPG4 CAR T cells target both differentiated SCCHN cells and SCCHN CICs. According to the cancer stem cell theory, CICs have to be completely eliminated for a therapy to be successful, since these cells play an important role in disease recurrence and in metastatic spread; and iii) CSPG4 is not detected in normal tissues except for activated pericytes in the tumor microenvironment. Therefore, immunotargeting of CSPG4 with CAR T cells is expected to inhibit neo-angiogenesis in the tumor microenvironment and to contribute to the elimination of SCCHN cells, even those with low or lack of CSPG4 expression. In recent studies, we have shown that CSPG4 CAR T cells are effective in eliminating CSPG4+ SCCHN cells in vitro under normoxic conditions. In addition, they significantly inhibit the growth of human SCCHN tumors in immunodeficient mice but they do not eradicate them. This proposal will test our working hypothesis that the hypoxia driven hostile microenvironment of SCCHN tumors causes CAR T cell dysfunction and reduces SCCHN cell susceptibility to CAR T cell mediated lysis. T cell plasticity allows us i) to restore CAR T cell function by disrupting PD-1/PD-L1 axis to counteract CAR T cell “exhaustion” and ii) to enhance viability and anti-tumor activity of CAR T cells by selectively increasing IL-15 level in the tumor microenvironment through the use of fusion proteins generated by linking IL-15 to anti-B7-H3 monoclonal antibody (mAb) HEK5. In addition, the susceptibility to CAR T cell mediated lysis of SCCHN cells will be restored by modulating anti-apoptotic molecule expression level through the inhibition with the small molecule sonidegib of hypoxia induced activation of Sonic Hedgehog Homolog pathway. The experiments will be performed in vitro utilizing SCCHN cell lines and in immunodeficient mice orthotopically grafted with both SCCHN cell lines and PDXs (some of which have already been established). The resulting information will determine whether the combinatorial strategy we have developed is effective in counteracting the hypoxia-related escape mechanisms utilized by SCCHN cells to avoid recognition and destruction by CSPG4 CAR T cells.

抽象的头颈鳞状细胞癌 (SCCHN) 的现有疗法疗效有限促使我们针对这种疾病设计一种新型有效的组合免疫疗法，在该策略中，T细胞。使用肿瘤抗原（TA）特异性嵌合抗原受体（CAR）工程设计作为效应器机制，因为这种方法允许 T 淋巴细胞特异性识别肿瘤和自我扩增我们选择硫酸软骨素蛋白聚糖 4 (CSPG4) 作为目标，因为 i) CSPG4 在约 60% 的 SCCHN 肿瘤中高表达，且每个肿瘤内的异质性有限；ii) CSPG4在分化的SCCHN细胞和具有高乙醛脱氢酶的SCCHN细胞上表达后者细胞被称为 ALDHbright 细胞，表现出癌症起始细胞 (CIC) 的特征，因为它们具有耐药性、表达干性基因并且在免疫缺陷小鼠中具有致瘤性。根据癌症干细胞，CSPG4 CAR T 细胞同时靶向分化的 SCCHN 细胞和 SCCHN CIC。根据细胞理论，必须完全消除 CIC 才能使治疗取得成功，因为这些细胞发挥着重要作用在疾病复发和转移扩散中发挥重要作用；以及 iii) 在正常组织中未检测到 CSPG4 除了肿瘤微环境中激活的周细胞，因此，用 CAR 免疫靶向 CSPG4。 T细胞有望抑制肿瘤微环境中的新血管生成并有助于消除 SCCHN 细胞，甚至那些 CSPG4 表达低或缺乏的细胞。在最近的研究中，我们发现CSPG4 CAR T细胞可以有效消除CSPG4+ SCCHN细胞此外，它们在常氧条件下体外还能显着抑制人 SCCHN 肿瘤的生长。免疫缺陷小鼠，但他们并没有消灭它们，这一提议将检验我们的工作假设： SCCHN 肿瘤的缺氧驱动的不利微环境导致 CAR T 细胞功能障碍并减少 SCCHN 细胞对 CAR T 细胞介导的裂解的敏感性使我们能够 i) 通过以下方式恢复 CAR T 细胞功能。破坏 PD-1/PD-L1 轴以抵消 CAR T 细胞“耗竭”，以及 ii) 增强活力和抗肿瘤作用通过使用选择性增加肿瘤微环境中的 IL-15 水平来增强 CAR T 细胞的活性通过将 IL-15 与抗 B7-H3 单克隆抗体 (mAb) HEK5 连接而生成的融合蛋白。通过调节抗凋亡分子可恢复对 CAR T 细胞介导的 SCCHN 细胞裂解的敏感性通过用小分子sonidegib抑制缺氧诱导的Sonic激活的表达水平 Hedgehog 同源途径将利用 SCCHN 细胞系在体外进行。免疫缺陷小鼠原位移植 SCCHN 细胞系和 PDX（其中一些已经由此产生的信息将决定我们是否拥有组合策略开发的可有效抵消 SCCHN 细胞利用的缺氧相关逃逸机制，以避免 CSPG4 CAR T 细胞的识别和破坏。