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.

抽象的可用疗法对头颈部鳞状细胞癌的效率有限（SCCHN）的效率有限促使我们为该疾病设计了一种新型有效的组合免疫疗法。在此策略中，T细胞用肿瘤抗原（TA）特异性嵌合抗原受体（CAR）设计为效应器机制，因为这种方法允许特异性肿瘤识别和由于T淋巴细胞引起的自我扩增自我更新能力。我们选择了硫酸软骨素蛋白聚糖4（CSPG4）作为靶标，因为I） CSPG4在大约60％的SCCHN肿瘤中高度表达，每个肿瘤中的异质性有限。 ii） CSPG4在具有高醛脱氢酶的分化SCCHN细胞和SCCHN细胞上表达活动。后者称为Aldhbright细胞，显示癌症启动的特征（CICS），由于它们具有耐药性，因此具有表达干性基因，并且在免疫缺陷的小鼠中是肿瘤的。所以， CSPG4 CAR T细胞靶向分化的SCCHN细胞和SCCHN CIC。根据癌症的茎细胞理论，必须完全消除CICS才能成功，因为这些细胞起着在疾病复发和转移性扩散中的重要作用；和iii）在正常组织中未检测到CSPG4 除了肿瘤微环境中活化的周细胞。因此，用汽车对CSPG4进行免疫靶向预计T细胞会抑制肿瘤微环境中的新血管生成，并有助于出现 SCCHN细胞，即使是低或缺乏CSPG4表达的细胞。在最近的研究中，我们表明CSPG4 CAR T细胞有效消除CSPG4+ SCCHN细胞在常氧条件下的体外。此外，它们显着抑制了人类SCCHN肿瘤在免疫缺陷的小鼠，但不会消除它们。该建议将检验我们的工作假设 SCCHN肿瘤的低氧驱动敌对微环境会导致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。 T细胞可塑性允许我们i）通过破坏PD-1/PD-L1轴以抵消CAR T细胞“疲惫”和II），以增强生存能力和抗肿瘤通过在肿瘤微环境中选择性地提高IL-15水平，通过使用通过将IL-15与抗B7-H3单克隆抗体（MAB）HEK5联系起来而产生的融合蛋白。另外，对CAR T细胞介导的SCCHN细胞裂解的敏感性将通过调节抗凋亡分子来恢复通过抑制小分子Sonidegib的表达水平，缺氧诱导的声音激活刺猬同源道。实验将在体外使用SCCHN细胞系和免疫缺陷的小鼠原位植入了SCCHN细胞系和PDX（其中一些已经建立）。由此产生的信息将确定我们是否有组合策略开发可有效抵消SCCHN细胞使用的缺氧相关的逃生机制 CSPG4 CAR T细胞的识别和破坏。