Functional characterization of novel oncogenic loci driving progression and immune response in gastrointestinal cancer

驱动胃肠癌进展和免疫反应的新型致癌位点的功能特征

• 批准号: 10746908
• 负责人: Yuan-Hung Lo
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746908
• 关键词: 3-Dimensional; ARID1A gene; Air; Anatomy; Automobile Driving; Biological; Biology; CRISPR interference; CRISPR-mediated transcriptional activation; CRISPR/Cas technology; Cancer Patient; Candidate Disease Gene; Cells; Chromosomal Instability; Coculture Techniques; Coupled; Cytotoxic Chemotherapy; Data; Detection; Disease; Early Diagnosis; Ecosystem; Engineering; Epithelium; Event; Evolution; Exhibits; Experimental Models; Gene Expression; Generations; Genes; Genetic; Genetic Engineering; Genome Stability; Genomic approach; Genomics; Genotype; Goals; Human; Human Herpesvirus 4; Immune; Immune Evasion; Immune response; Immune system; Immunologic Deficiency Syndromes; Immunology; Immunooncology; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; Inflammatory; Liquid substance; MLH1 gene; Malignant Neoplasms; Malignant neoplasm of gastrointestinal tract; Measures; Mediating; Mentors; Methodology; Methods; Microsatellite Instability; Modeling; Molecular; Monitor; Mutate; Mutation; Natural Immunity; Natural Killer Cells; Normal tissue morphology; Oncogenes; Oncogenic; Oncology; Organoids; Patients; Phase; Phenotype; Physiological; Population; Predisposition; Proteins; Reaction; Regulation; Research; Research Personnel; Resistance; Resistance development; Role; Series; Shapes; Somatic Mutation; System; T-Lymphocyte; TP53 gene; Technology; Testing; The Cancer Genome Atlas; Therapeutic; Training; Tumor Immunity; Tumor-Infiltrating Lymphocytes; Work; Xenograft procedure; adaptive immunity; cancer cell; cancer immunotherapy; cancer subtypes; cancer surgery; cancer therapy; career; cell killing; conventional therapy; gastric organoids; gastric tumorigenesis; genetic approach; genetic manipulation; genome editing; immune cell infiltrate; immune checkpoint blockade; immunoregulation; improved; in vitro Model; in vivo; insight; malignant stomach neoplasm; mouse model; multimodality; mutant; neoantigens; neoplasm immunotherapy; neoplastic cell; next generation; novel; novel therapeutics; patient subsets; pharmacologic; preservation; response; skills; success; tool; treatment response; tumor; tumor immunology; tumor microenvironment; tumor-immune system interactions; tumorigenesis

PROJECT SUMMARY/ABSTRACT Gastric cancer (GC) is one of the most common and lethal cancers worldwide. GC surgery is highly morbid, and responses to the limited array of treatment options are poor. There is hope that recent genomic sequencing data can be leveraged to develop newer, improved molecular therapies for GC, but rigorous mechanistic testing is still needed to validate the therapeutic potential of targeting any newly proposed oncogenes. Immunotherapy is an exciting new therapy that has revolutionized oncology and shows tremendous potential. In contrast to cytotoxic chemotherapies, which exhibit fractional killing invariably leading to resistance, immune cells can infiltrate almost all anatomic sites to recognize and completely eliminate malignant cells in primary and wide-spread metastatic disease. However, the immune system’s full anti-tumor killing potential can be restricted by evasive measures by the tumor and/or intrinsic immunosuppressive mechanisms that limit collateral damage to normal tissues during anti-tumor inflammatory reactions. In GC, little is known about how cancer cells evade the system, and studies investigating the molecular mechanisms underlying tumor- immune interactions have been limited by a lack of physiologically relevant in vitro human systems where state-of-the-art genetic approaches can be applied. These mechanisms are important because they would be essential to our understanding of GC tumorigenesis and the regulation of immunotherapeutic responses. Such mechanistic insight on the immune system to GC is fundamental and significant to advance and improve GC therapies. In this proposal, we utilize a series of CRISPR/Cas9 genome editing tools to create novel forward genetically engineered models of the four major GC subtypes as defined by The Cancer Genome Atlas project, including chromosomal instability (CIN), genomic stability (GS), microsatellite instability (MSI) and Epstein-Barr virus-associated (EBV) in primary 3D human gastric organoids (Aim1 and Aim2). In a parallel translational aim, we propose to use a second-generation patient-derived organoid model that allows tumor and stroma to be preserved alongside each other to study interactions between tumor cells and their veritable ecosystem of cohabiting immune cells in primary human gastric cancer (Aim 3). The overall goal of this project is to investigate how genetic alterations contribute to gastric tumorigenesis and immunotherapeutic responses using synergistic next-generation in vivo and in vitro models. Collectively, the results of this project will provide new insights into fundamental aspects of the molecular mechanisms underlying the tumor-immune interaction and enhance current GC immunotherapies. A team of expert mentors, advisors and collaborators will train Dr. Lo in new methods that are critical to the success of this research. The combination of mentoring support, skills, and data obtained in the K99 phase will provide Dr. Lo a springboard to achieving independence as an investigator in the R00 phase and beyond.

项目摘要/摘要 胃癌是世界范围内最常见、最致命的癌症之一。GC手术是高度病态的， 而且对有限的治疗选择的反应很差。人们希望最近的基因组测序 可以利用数据来开发更新的、改进的GC分子疗法，但需要进行严格的机械测试 仍然需要验证靶向任何新提出的癌基因的治疗潜力。 免疫疗法是一种令人兴奋的新疗法，它使肿瘤学发生了革命性的变化，并显示出巨大的 潜力。与细胞毒性化疗不同的是，细胞毒性化疗表现出一定程度的杀伤力， 免疫细胞可以渗透到几乎所有的解剖部位，识别和完全消除恶性细胞。 原发和广泛传播的转移性疾病。然而，免疫系统的全部抗肿瘤杀伤潜力可以 被肿瘤的回避措施和/或限制的固有免疫抑制机制所限制 在抗肿瘤炎症反应中对正常组织的附带损害。在GC中，人们对此知之甚少 癌细胞逃避这个系统，研究肿瘤的分子机制-- 由于体外人体系统缺乏生理上的相关性，免疫相互作用受到限制。 在那里可以应用最先进的遗传方法。这些机制很重要，因为它们 对于我们理解GC肿瘤的发生和免疫治疗的调节是必不可少的 回应。这种对免疫系统的机制洞察对于GC的发展是基本的和有意义的 并改进GC疗法。在这个方案中，我们利用一系列CRISPR/Cas9基因组编辑工具来创建 癌症基因组定义的四种主要GC亚型的新的正向基因工程模型 Atlas计划，包括染色体不稳定(CIN)、基因组稳定(GS)、微卫星不稳定(MSI)和 原代3D人胃器官(Aim1和AIM2)中的EB病毒相关(EBV)并行地 翻译目标，我们建议使用第二代患者衍生的器官模型，允许肿瘤和 间质将被并排保存，以研究肿瘤细胞和它们之间的相互作用 人类原发胃癌的共生免疫细胞生态系统(目标3)。这个项目的总体目标是 是研究基因改变如何影响胃肿瘤的发生和免疫治疗反应 采用协同下一代体内和体外模型。总的来说，该项目的成果将提供 对肿瘤-免疫相互作用分子机制基本方面的新见解 并加强目前的GC免疫疗法。 一个由专家导师、顾问和合作者组成的团队将培训罗博士学习对以下方面至关重要的新方法 这项研究的成功。指导支持、技能和在K99阶段获得的数据的组合 将为罗博士提供一个跳板，在R00阶段及以后成为一名独立的研究人员。