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: 10450179
• 负责人: Yuan-Hung Lo
• 金额: $ 17.17万
• 依托单位: STANFORD UNIVERSITY
• 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 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10450179
• 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; Ecosystem; Engineering; Epithelial; 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; Immunology; Immunooncology; Immunotherapeutic agent; Immunotherapy; In Vitro; 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; Pharmacology; 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; Tumor-infiltrating immune cells; Work; Xenograft procedure; adaptive immunity; base; 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 checkpoint blockade; 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; 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.

项目总结/摘要 胃癌是世界范围内最常见、最致命的恶性肿瘤之一。胃癌手术是非常病态的， 对有限的治疗方案的反应很差。最近的基因组测序有望 这些数据可以用来开发更新的、改进的GC分子疗法， 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，它彻底改变了肿瘤学， 潜力与细胞毒性化疗相反，细胞毒性化疗表现出一定程度的杀伤作用，导致耐药性， 免疫细胞可以渗透到几乎所有的解剖部位，以识别并完全消除恶性细胞， 原发性和广泛转移性疾病。然而，免疫系统的全部抗肿瘤杀伤潜力可以 通过肿瘤的逃避措施和/或限制免疫的内在免疫抑制机制来限制 在抗肿瘤炎症反应期间对正常组织的附带损害。在GC中，我们对如何使用 癌细胞逃避系统，研究调查肿瘤的分子机制- 免疫相互作用由于缺乏生理学相关的体外人体系统而受到限制 最先进的遗传学方法可以应用的地方。这些机制很重要，因为它们 对于我们理解胃癌的发生和免疫调节是至关重要的。 应答这种对免疫系统对GC的机制性认识是基础性的，对推进 并改善GC疗法。在这个提议中，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亚型的新型正向基因工程模型 Atlas项目，包括染色体不稳定性（CIN）、基因组稳定性（GS）、微卫星不稳定性（MSI）和 原发性3D人类胃类器官（Aim 1和Aim 2）中的EB病毒相关（EBV）。以平行 翻译的目的，我们建议使用第二代患者来源的类器官模型，允许肿瘤和 相互保存，以研究肿瘤细胞与其真正的肿瘤细胞之间的相互作用。 原发性人胃癌中同居免疫细胞的生态系统（目的3）。本项目的总体目标 是研究基因改变如何影响胃癌的发生和免疫反应 使用协同的下一代体内和体外模型。总的来说，该项目的结果将提供 对肿瘤-免疫相互作用的分子机制的基本方面的新见解 并增强目前的GC免疫疗法。 一个由专家导师、顾问和合作者组成的团队将对罗博士进行新方法的培训， 这项研究的成功。在K99阶段获得的指导支持、技能和数据的组合 将为Lo博士提供一个跳板，使其在R 00阶段及以后实现作为研究者的独立性。