权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of novel metastatic mouse models that recapitulate the major immune contexts of human colon cancer

开发新的转移性小鼠模型来概括人类结肠癌的主要免疫环境

基本信息

批准号：
10063490
负责人：
TYLER E. JACKS
金额：
$ 32.88万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10063490
关键词：
Address Adoptive Transfer Alleles Antibodies Antigens Benchmarking CRISPR/Cas technology Cancer Model Chickens Clinical Clinical Trials Colon Colon Carcinoma Colonoscopy Colorectal Cancer Comparative Study DNA Development Disease Dissection Engineering Enterobacteria phage P1 Cre recombinase FDA approved Genes Genetic Genetic Engineering Goals Guide RNA Histology Human Immune Immune Evasion Immune checkpoint inhibitor Immune response Immunotherapy Injections Kinetics Knock-out Lentivirus Liver Lung MSH2 gene Mediating Methodology Methods Mismatch Repair Mismatch Repair Deficiency Modeling Mus Mutation Neoplasm Metastasis Organoids Ovalbumin Patients Preclinical Testing Predisposition Primary Neoplasm Refractory Research Solid Neoplasm Somatic Mutation Standardization Stimulus T cell response T cell therapy T-Lymphocyte Techniques Testing Tissues Translational Research Treatment outcome Tumor Antigens Tumor-infiltrating immune cells analog anti-CTLA4 antibodies anti-PD-1 antigen-specific T cells base cancer therapy cancer type chemotherapy clinical predictors colon cancer patients design exhaustion immune checkpoint immune checkpoint blockade immunogenic immunogenic cell death immunogenicity immunotherapy trials improved inducible gene expression innovation model development mouse genetics mouse model mutant neoantigens next generation sequencing novel organoid transplantation patient population patient subsets pre-clinical preclinical trial programmed cell death protein 1 response transplant model tumor tumor-immune system interactions

项目摘要

The purpose of this proposal is to develop novel mouse models of colorectal cancer (CRC) with appropriate immune responses. These models have been designed to address limitations present in current models of CRC in order to enhance their suitability for translational research in immunotherapy. Immune checkpoint inhibitors have revolutionized treatment of solid tumors, and brought to light the critical importance of tumor immune context in treatment outcome. CRC with DNA mismatch repair (MMR) deficiency is characterized by a high burden of somatic mutations, increased T cell infiltration, and a favorable response to checkpoint blockade. Unfortunately, the majority of CRC has a lower mutational burden and is refractory to these treatments. Preclinical mouse models are powerful platforms for investigating the factors underlying response to immunotherapy. However, no single model faithfully recapitulates primary tumor development in the colon microenvironment, metastatic dissemination to the liver and lung, and the major immune contexts underlying variability in immunotherapy response in human CRC. To address these significant translational deficiencies, we will use an innovative technique employing colonoscopy-guided sub-mucosal injection of lentivirus or tumor organoids to induce focal autochthonous and orthotopic tumors in the colon that readily metastasize. In Aim 1, we will engineer a model that modulates tumor immunogenicity through inducible expression of a model antigen. We will dissect the features of the induced anti-tumor T cell response and investigate the utility of this model for testing adoptive T cell therapy by transferring antigen-specific T cells. To potentiate adoptive T cell therapy and mirror ongoing clinical trials in humans, we will assess the efficacy of CRISPR-Cas9-mediated deletion of immune checkpoints in T cells prior to transfer. In Aim 2, we will model immunotherapy-responsive CRC by targeting the essential DNA MMR genes Msh2 and Mlh1 and use next-generation sequencing to characterize the mutational landscapes of resulting MMR-deficient versus proficient tumors. In Aim 3, we will perform preclinical trials of immune checkpoint blockade in these models to explore their ability to recapitulate the responses of human CRC patient populations. We will also test a novel combination of immunogenic chemotherapy and checkpoint blockade, based on the hypothesis that immunogenic cell death may sensitize tumors with low mutational burden or minimal pre-existing T cell involvement to immune attack. This strategy is aimed at improving treatment for the majority of CRC patients, whose tumors are non-immunogenic and non- responsive to immunotherapy. The overarching goal of this research plan is to develop and benchmark a set of highly comparable CRC models that will be used to address why only a fraction of patients respond to immunotherapy. The proposed strategy is innovative in that it uses cutting-edge methods in mouse genetic engineering and cancer modeling to capture critical features of human CRC. This research will also include deep characterization of the immune microenvironment in these models and a comparison to humans.

本提案的目的是开发新的结直肠癌（CRC）小鼠模型，免疫反应。这些模型旨在解决当前模型中存在的局限性， CRC，以提高其在免疫治疗转化研究中的适用性。免疫检查点抑制剂已经彻底改变了实体瘤的治疗，并揭示了肿瘤抑制剂的关键重要性。治疗结果中的免疫背景。CRC与DNA错配修复（MMR）缺陷的特点是，体细胞突变的高负荷，T细胞浸润增加，以及对检查点的有利反应封锁不幸的是，大多数CRC具有较低的突变负担，并且对这些突变是难治的。治疗。临床前小鼠模型是研究反应潜在因素的强大平台免疫疗法然而，没有一个单一的模型忠实地重演原发性肿瘤的发展，在结肠微环境，转移性扩散到肝脏和肺，以及主要的免疫背景下，人CRC中免疫治疗应答的变异性。为了解决这些重大的翻译缺陷，我们将使用一种创新的技术，采用结肠镜引导下粘膜下注射慢病毒或肿瘤，类器官诱导结肠中容易转移的局灶性原位和原位肿瘤。在目标1中，我们将设计一种模型，通过诱导表达一种模型来调节肿瘤免疫原性，抗原的我们将剖析诱导的抗肿瘤T细胞应答的特征，并研究这种应答的效用。用于通过转移抗原特异性T细胞来测试过继性T细胞疗法的模型。为了增强过继性T细胞为了评估CRISPR-Cas9介导的治疗和反映正在进行的人类临床试验，我们将评估CRISPR-Cas9介导的治疗的有效性。在转移之前删除T细胞中的免疫检查点。在目标2中，我们将模拟免疫治疗响应通过靶向必需的DNA MMR基因Msh 2和Mlh 1，并使用下一代测序，表征所产生的MMR缺陷型肿瘤与成熟型肿瘤的突变景观。在目标3中，我们在这些模型中进行免疫检查点阻断的临床前试验，以探索它们重演人类CRC患者群体的反应。我们还将测试一种新的免疫原性化疗和检查点阻断，基于免疫原性细胞死亡可能使具有低突变负荷或最小的预先存在的T细胞参与免疫攻击的肿瘤。这一战略是旨在改善大多数CRC患者的治疗，其肿瘤是非免疫原性的，对免疫疗法有反应。本研究计划的总体目标是开发一套高度可比的CRC模型，将用于解决为什么只有一小部分患者对免疫疗法。该策略的创新之处在于它使用了小鼠遗传学中的尖端方法，工程和癌症建模，以捕获人类CRC的关键特征。这项研究还将包括这些模型中免疫微环境的深入表征以及与人类的比较。