Unbiased discovery and mechanistic interrogation of genetic drivers of malignant transformation

恶性转化遗传驱动因素的公正发现和机械询问

• 批准号: 10083636
• 负责人: Adan H. Codina
• 金额: $ 0.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2020-12-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10083636
• 关键词: Acute; Affect; Animals; Antigens; Area; Automobile Driving; Biological Assay; C57BL/6 Mouse; CRISPR screen; Catalogs; Cause of Death; Cell Differentiation process; Cell Line; Cells; Cessation of life; Clinical; Clinical Data; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Cyclic AMP-Dependent Protein Kinases; Data; Extracellular Matrix; Genes; Genetic; Genetic Transcription; Genetically Engineered Mouse; Genotype; Hepatocarcinogenesis; Hepatocyte transplantation; Histopathology; Homeostasis; Immune; Immune system; Immunocompetence; Immunocompetent; Immunologic Markers; Inflammation; Inflammatory Response; Interleukin-6; Investigation; Knock-out; Knowledge; Liver; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of liver; Mediating; Mediator of activation protein; Medical; Modeling; Molecular; Molecular Biology; Mus; Mutation; Myeloid-derived suppressor cells; NU/NU Mouse; Oncogenes; Outcome; Pathway interactions; Phenotype; Play; Population; Primary carcinoma of the liver cells; Process; Proteins; Proteomics; Rag1 Mouse; Regulation; Regulatory T-Lymphocyte; Role; Sampling; Signal Transduction; Signaling Molecule; Somatic Mutation; System; Testing; Tissues; Transcript; Tumor Escape; Tumor Tissue; Tumor-Infiltrating Lymphocytes; Tumor-infiltrating immune cells; United States; Validation; Work; Xenograft Model; cancer genome; cell type; cellular pathology; clinically relevant; cytotoxic CD8 T cells; experimental study; genome-wide; granulocyte; human disease; in vivo; insight; interest; molecular scale; mouse model; mutant; preservation; profiles in patients; prognostic; temporal measurement; therapeutic development; transcriptome sequencing; tumor; tumor growth; tumor microenvironment; tumor progression; tumor xenograft; tumor-immune system interactions; tumorigenesis

Project Summary Cancer is a leading cause of death in the United States, causing hundreds of thousands of deaths each year. For the majority of these cases, tumors develop as a result of accumulated somatic mutations, of which thousands of clinically relevant combinations exist. To better understand this process the greater scientific and medical community has performed large-scale molecular profiling of patient samples. Such studies have identified a comprehensive catalog of molecular alterations in the cancer genome. However, the functional roles of many of these cancer genes still need to be explicitly tested in controlled experimental settings. Moreover, even fewer genes have been functionally probed for how they affect the tumor micro-environment (TME), an area of increasing interest and importance. It is therefore imperative to systematically and quantitatively assess the contribution of each gene on tumor progression and influence on the TME. To this end we leveraged an unbiased genome-scale CRISPR-Cas9 screen and identified and validated drivers of tumorigenesis in the in the context of liver hepatocellular carcinoma or (HCC). Of these drivers, Prkar1a, demonstrated robust tumor growth in validation experiments when knocked out in non-tumorigenic hepatocytes and transplanted into Nu/Nu mice. Additionally, Prkar1a’s role in liver cancer, or cancer more generally is not well understood. Therefore, we chose this gene for further mechanistic interrogation. In this proposal we will further probe the importance of Prkar1a for their role in liver tumorigenesis through two specific aims: AIM 1: Interrogate the molecular and cellular mechanism of ECM remodeling by Prkar1a AIM2: Develop clinically relevant models and dissect the immune landscape of Prkar1a-/- tumors. Together, these experiments will elucidate how Prkar1a knockout alters the TME, and promotes tumorigenesis in models of HCC. These findings will provide fundamental insights into Prkar1a-/- tumorigenesis, and assist in therapeutic development for these tumors.

项目摘要 癌症是美国的主要死因，每年造成数十万人死亡。 对于这些病例中的大多数，肿瘤是由于累积的体细胞突变而发展的，其中 存在数千种临床相关的组合。为了更好地理解这一过程， 医学界已经对患者样品进行了大规模的分子分析。这类研究 确定了癌症基因组中分子改变的全面目录。然而，功能 许多这些癌症基因的作用仍然需要在受控的实验环境中进行明确的测试。 此外，甚至更少的基因已经被功能性地探测了它们如何影响肿瘤微环境 (TME)这是一个越来越受关注和重要的领域。因此，必须有系统地和 定量评估每个基因对肿瘤进展的贡献和对TME的影响。本 最后，我们利用无偏见的基因组规模CRISPR-Cas9筛选，并确定和验证了 在肝细胞癌或（HCC）的背景下的肿瘤发生。在这些司机中，Prkar 1a， 在验证实验中，当在非致瘤性肝细胞中敲除时， 并移植到Nu/Nu小鼠中。此外，Prkar 1a在肝癌或更普遍的癌症中的作用不是 很好理解。因此，我们选择了这个基因进行进一步的机制研究。在本提案中，我们将 通过两个特定目的进一步探索Prkar 1a在肝肿瘤发生中的作用的重要性：AIM 1： Prkar 1a AIM 2对ECM重塑的分子和细胞机制的探讨 临床相关的模型和解剖Prkar 1a-/-肿瘤的免疫景观。所有这些 实验将阐明Prkar 1a基因敲除如何改变TME，并促进肿瘤模型中的肿瘤发生。 HCC。这些发现将为Prkar 1a-/-肿瘤发生提供基本见解，并有助于治疗 这些肿瘤的发展。