Cancer Cell Intrinsic Interferon-I pathway Activation by Fractionated Radiation

分段放射激活癌细胞内源性干扰素-I 通路

基本信息

批准号：
9207065
负责人：
Sandra Demaria
金额：
$ 38.77万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-19 至 2020-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9207065
关键词：
4T1 Acute Address Affect Aftercare Antigens Antineoplastic Agents Bone Marrow Breast Cancer Cell CD8-Positive T-Lymphocytes Cancer Patient Cancer Survivor Cancer Vaccines Carcinoma Cell Death Cells Chimera organism Clinic Cyclic GMP Cytotoxic T-Lymphocyte-Associated Protein 4 DNA Data Dendritic Cells Dendritic cell activation Development Dose Dose Fractionation Epitopes Functional disorder Gap Junctions Gene Activation Generations Genes Hematopoietic Human IFNAR1 gene Immune Immune checkpoint inhibitor Immune response Immunocompetent Immunologic Adjuvants Immunosuppressive Agents Immunotherapy Implant In Situ Individual Infiltration Interferon Type I Interferons Ionizing radiation Irradiated tumor Knowledge Ligands MC38 MCF7 cell MDA MB 231 Malignant Epithelial Cell Mediating Modality Modeling Molecular Mus Mutate Mutation Myeloid Cells Neoplasm Metastasis Neoplastic Epithelial Cell Pathway interactions Patients Phenotype Pre-Clinical Model Proteins Radiation Radiation therapy Recruitment Activity Regimen Role Signal Transduction T cell response T-Cell Activation T-Lymphocyte Testing Therapeutic Tumor Antigens Tumor Immunity Vaccinated Vaccination Vaccines Virus Diseases Wild Type Mouse Work advanced disease cancer cell cancer therapy carcinogenesis chemotherapy experimental study immune checkpoint immune checkpoint blockade immunogenic improved in vitro testing in vivo knock-down neoplastic cell novel novel strategies overexpression pre-clinical public health relevance receptor response sensor small hairpin RNA success tumor tumor microenvironment

项目摘要

DESCRIPTION (provided by applicant): Recent evidence indicates that the most powerful anti-tumor T cells recognize neoantigens derived from unique mutated proteins expressed by an individual tumor, suggesting that precision vaccination is required to induce effective anti-tumor immunity in cancer patients. Importantly, such anti-tumor T cells cause tumor regression at advanced disease stages upon therapeutic immune checkpoints blockade. We have pioneered studies exploring the use of local tumor radiotherapy (RT) as a means to generate an in situ individualized vaccine. We were the first to demonstrate in a pre-clinical model that RT sensitizes unresponsive tumors to CTLA-4 blockade by inducing T cells specific for endogenous tumor antigens. Emerging evidence by us and others suggests a similar effect of RT in the clinic. However, lack of knowledge about the mechanisms involved precludes rapid progress towards the effective use of RT as an immune adjuvant. One critical unanswered question is whether dose and fractionation affect RT ability to elicit anti-tumor immune responses. We have previously found in two mouse carcinomas that generation of an in situ vaccine synergistic with anti-CTLA-4 treatment in inducing immune-mediated regression of irradiated and synchronous non-irradiated tumors (abscopal effect) was achieved by RT given in 3 fractions of 8 Gy but not by a single 20 Gy dose, suggesting that the RT regimen employed is critical. We now have data supporting the hypothesis that carcinoma cell-intrinsic activation of type I interferon (IFN-I) pathway by RT is required to generate an in situ vaccine, suggesting that RT triggers canonical defense pathways in neoplastic epithelial cells that mimic a viral infection. Our data also indicat that fractionated (FRT) but not single dose (SDRT) radiation can accomplish this via activation of the cytosolic DNA sensor cyclic GMP-AMP synthase (cGAS) and downstream stimulator of IFN genes (STING) within the irradiated cancer cells. To test the above hypothesis several mouse and human carcinoma cells will be employed to determine which RT dose/fractionation activates cGAS/STING. Next, in vivo experiments using cancer cells with selective knockdown of cGAS or STING implanted in syngeneic immunocompetent wild type, cGAS-deficient, and STING-deficient mice will be performed to determine the role of this pathway in RT-mediated induction of anti-tumor T cells that mediate tumor regression and abscopal responses. To determine whether the effects of IFN-I produced by irradiated cancer cells are cancer cell autonomous or require signaling in host DC, IFNAR1-deficient mice will be used as tumor recipient. The relationship between cancer cell-derived IFN-I and DC recruitment to tumors will be established. Finally, we will investigate the mechanisms whereby RT activates the cGAS/STING pathway in cancer cells. Overall, data obtained will have important implications for a novel use of radiotherapy as a relatively simple and widely available modality for individualized tumor vaccination.

描述（由适用提供）：最近的证据表明，最强大的抗肿瘤T细胞识别由单个肿瘤表达的独特突变蛋白得出的新抗原，这表明需要精确疫苗接种以诱导癌症患者中有效的抗肿瘤免疫组织化学。重要的是，这种抗肿瘤T细胞在治疗性免疫化学检查点阻塞后在晚期疾病阶段引起肿瘤消退。我们的开创性研究探讨了局部肿瘤放射疗法（RT）作为生成原位个性化疫苗的一种手段。我们是第一个在临床前模型中证明的人，该模型通过诱导针对内源性肿瘤抗原的T细胞，从而感知到CTLA-4阻断的无反应性肿瘤。我们和其他人的新证据表明，RT在诊所中的影响相似。但是，缺乏有关涉及机制的知识阻碍了有效使用RT作为免疫辅助的快速进步。一个关键的未解决的问题是，剂量和分馏是否影响RT引起抗肿瘤免疫复杂的能力。以前，我们在两种小鼠癌中已经发现，在诱导的免疫介导的辐射和同步非辐照肿瘤的免疫介导的回归中，与抗CTLA-4处理的产生相结合（脱落效应）是由RT通过8 Gy给出的3 Gy的3个级分，而不是由RT给出的。现在，我们有数据支持以下假设：需要通过RT进行I型干扰素（IFN-I）途径的癌细胞中性激活，以生成原位疫苗，这表明RT触发了模仿病毒感染的肿瘤上皮细胞中的肿瘤上皮细胞中的典型防御途径。我们的数据还表明，分馏（FRT）但不单一剂量（SDRT）辐射可以通过激活胞质DNA传感器环状GMP-AMP合酶（CGA）和IFN基因（STING）的下游刺激剂的激活来实现这一目标。为了测试上述假设，将使用几只小鼠和人类癌细胞来确定哪种RT剂量/分级激活CGAS/STING。接下来，使用癌细胞进行选择性敲低的CGA或植入植入的合成性免疫能力的野生型，CGAS和刺激性小鼠的体内实验，以确定该途径在RT介导的抗肿瘤T细胞中的作用，以介导抗肿瘤T细胞介导的抗肿瘤T细胞介导的肿瘤肿瘤回归和露出响应。为了确定受辐照癌细胞产生的IFN-I的影响是癌细胞自主的还是需要在宿主DC中信号传导的，IFNAR1熟的小鼠将被用作肿瘤受体。将建立癌细胞衍生的IFN-I和DC募集到肿瘤之间的关系。最后，我们将研究RT激活癌细胞中CGA/STING途径的机制。总体而言，获得的数据将对放射疗法的新型使用作为一种相对简单且可广泛可用的方式，以实现个性化的肿瘤疫苗接种。