Unbiased discovery and mechanistic interrogation of genetic drivers of malignant transformation

项目摘要 癌症是美国的主要死亡原因，每年导致数十万人死亡。 对于这些病例中的大多数，肿瘤是由于积累的体细胞突变而发展的，其中 存在着数千种临床上相关的组合。为了更好地理解这一过程，更大的科学和 医学界已经对患者样本进行了大规模的分子图谱分析。这类研究已经 确定了癌症基因组中分子变化的全面目录。然而，功能性的 其中许多癌症基因的作用仍需要在对照实验环境中明确测试。 此外，从功能上研究它们如何影响肿瘤微环境的基因就更少了。 (TME)，一个日益引起人们兴趣和重要性的领域。因此，必须系统地和 定量评估每个基因对肿瘤进展的贡献和对TME的影响。对这件事 最终，我们利用了无偏见的基因组规模CRISPR-Cas9筛查，并确定和验证了 在肝细胞癌或(肝细胞癌)的背景下的肿瘤发生.在这些司机中，Prkar1a， 在验证实验中，当在非致瘤肝细胞中被敲除时，显示出强劲的肿瘤生长 并移植到NU/NU小鼠体内。此外，Prkar1a在肝癌中的作用，或者更广泛地说，在癌症中不是 很好理解。因此，我们选择了这个基因进行进一步的机械审讯。在本提案中，我们将 通过两个特定的目标进一步探讨Prkar1a在肝脏肿瘤发生中的作用：目标1： Prkar1a AIM2重塑ECM的分子和细胞机制研究进展 临床相关模型，并剖析Prkar1a-/-肿瘤的免疫格局。加在一起，这些 实验将阐明Prkar1a基因敲除如何改变TME，并促进肿瘤发生 肝细胞癌。这些发现将为Prkar1a-/-肿瘤的发生提供基本的见解，并有助于治疗 这些肿瘤的发展。

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.