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

• 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
• 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

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.

摘要 头颈部鳞状细胞癌（SCCHN）现有治疗的疗效有限， 促使我们设计一种新的有效的组合免疫治疗这种疾病。在这种策略中，T细胞 用肿瘤抗原（TA）-特异性嵌合抗原受体（CAR）工程化的细胞用作效应子 由于这种方法允许肿瘤识别的特异性和由于T淋巴细胞的自我扩增， 自我更新能力我们选择硫酸软骨素蛋白聚糖4（CSPG 4）作为靶标，因为i） CSPG 4在约60%的SCCHN肿瘤中高度表达，每个肿瘤内的异质性有限; ii） CSPG 4在分化的SCCHN细胞和具有高醛脱氢酶的SCCHN细胞上均表达 活动后一种细胞被称为ALDHbright细胞，显示出癌症起始细胞（CIC）的特征， 因为它们具有抗药性、表达干细胞基因且在免疫缺陷小鼠中具有致瘤性。因此，我们认为， CSPG 4 CAR T细胞靶向分化的SCCHN细胞和SCCHN CIC。根据癌症干细胞 根据细胞理论，CIC必须完全消除才能使治疗成功，因为这些细胞在治疗中起着重要的作用。 在疾病复发和转移扩散中起重要作用;和iii）在正常组织中未检测到CSPG 4 除了肿瘤微环境中的活化周细胞。因此，用CAR免疫靶向CSPG 4， 预期T细胞抑制肿瘤微环境中的新血管生成，并有助于消除肿瘤微环境中的肿瘤细胞。 SCCHN细胞，即使是那些CSPG 4表达低或缺乏的细胞。 在最近的研究中，我们已经表明CSPG 4 CAR T细胞在消除CSPG 4 + SCCHN细胞中是有效的。 在常氧条件下体外培养。此外，它们显著抑制人SCCHN肿瘤的生长， 免疫缺陷小鼠，但他们不能根除它们。这项建议将检验我们的工作假设， 缺氧驱动的SCCHN肿瘤的不利微环境导致CAR T细胞功能障碍并减少SCCHN 细胞对CAR T细胞介导的裂解的易感性。T细胞可塑性允许我们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”，和ii）增强存活力和抗肿瘤活性， CAR T细胞的活性，通过选择性增加肿瘤微环境中的IL-15水平，通过使用 通过将IL-15连接至抗B7-H3单克隆抗体（mAb）HEK 5产生的融合蛋白。此外该 对CAR T细胞介导SCCHN细胞裂解的易感性将通过调节抗凋亡分子 通过小分子sonidegib抑制缺氧诱导的Sonic激活， Hedgehog Homolog途径。实验将利用SCCHN细胞系在体外进行，并在 原位移植SCCHN细胞系和PDX的免疫缺陷小鼠（其中一些已经 成立）。得到的信息将决定我们的组合策略 开发的是有效的抵消缺氧相关的逃逸机制所利用的SCCHN细胞，以避免 CSPG 4 CAR T细胞的识别和破坏。