Molecular Dissection of Lung Cancer Progression and Metastasis

描述（由申请人提供）：肺癌是美国男性和女性癌症死亡的主要原因，仅在这个国家每年就有超过155，000名患者死亡。有几个因素导致肺癌患者的预后不佳，但是，与大多数实体瘤一样，癌细胞离开原发肿瘤并建立不可手术转移的能力是成功治疗的主要障碍。因此，转移代表了一个主要的临床挑战，这是由尚未充分了解的细胞状态改变驱动的。这项建议 使用新的方法来揭示肺癌进展和转移级联的每一步的分子和细胞变化。我们将使用一个基因工程肺腺癌小鼠模型，该模型概括了人类肺腺癌的遗传改变和组织学进展。在Aim 1中，我们将在体内功能性地询问肺癌转移促进和抑制基因，通过使用慢病毒载体系统直接表达肺癌转移的合理候选调节因子。我们将联合收割机这些工具与肺腺癌的基因工程小鼠模型相结合，该模型结合了癌细胞的荧光标记，以量化转移级联的每一步。在Aim2中，我们将通过荧光激活细胞分选从原发性肿瘤和转移瘤中分离肺腺癌细胞，以揭示定义恶性进展每个阶段的基因表达谱。直接转移能力的基因表达变化将有助于确定恶性疾病状态的潜在生物标志物 揭示原发肿瘤及其相关器官转移中基因表达的整体关系。在Aim3中，我们将通过人肺腺癌中候选基因表达与临床结局的相关性，并使用定义的肺上皮细胞转化系统来评估细胞培养和体内的基因功能，分析和功能解剖人肺腺癌进展过程中的基因功能。鉴于转移性癌症的巨大临床影响以及目前在理解这种疾病状态的分子基础方面的差距，临床实践和患者结局将受到任何新疗法的极大影响，这些新疗法可能来自我们提出的分析中获得的基础知识。通过结合定量方法和强大的体内方法，我们希望揭示控制肿瘤进展和转移扩散的一般原则，并最终发现癌症进展连续体中的新治疗靶点。

DESCRIPTION (provided by applicant): Lung cancer is the leading cause of cancer deaths in both men and women in the United States, with over 155,000 patients dying each year in this country alone. Several factors contribute to the poor outcome of lung cancer patients, but, as in most solid tumors, the ability of cancer cells to leave the primary tumor and establish inoperable metastases is a major impediment to successful therapy. Metastasis thus represents a major clinical challenge that is driven by as yet poorly understood cell state alterations. This proposal uses novel methods to uncover the molecular and cellular changes that underlie lung cancer progression and each step of the metastatic cascade. We will use a genetically-engineered lung adenocarcinoma mouse model that recapitulates the genetic alterations and histological progression of human lung adenocarcinoma. In Aim1, we will functionally interrogate lung cancer metastasis-promoting and -inhibiting genes in vivo, by using a lentiviral vector system to directly express rational candidate regulators of lung cancer metastasis in developing tumors in vivo. We will combine these tools with a genetically-engineered mouse model of lung adenocarcinoma which incorporates fluorescent marking of cancer cells to allow quantification of each step of the metastatic cascade. In Aim2, we will isolate lung adenocarcinoma cells from primary tumors and metastases by fluorescence activated cell-sorting to uncover gene expression profiles that define each stage of malignant progression. Gene expression changes that direct metastatic ability will help define potential biomarkers of the malignant disease state and reveal the relationship of global gene expression in primary tumors and their related metastases in different organs. In Aim3, we will analyze and functionally dissect gene function during human lung adenocarcinoma progression, through the correlation of candidate gene expression in human lung adenocarcinoma with clinical outcome and use of a defined lung epithelial cell transformation system to assess gene function both in cell culture and in vivo. Given the immense clinical impact of metastatic cancer and the current gap in understanding the molecular underpinnings of this disease state, both clinical practice and patient outcome would be greatly impacted by any new therapies that might result from the fundamental knowledge gained from our proposed analyses. By combining quantitative methods and powerful in vivo methods, we hope to uncover general principles that govern tumor progression and metastatic spread and ultimately uncover novel therapeutic targets across the continuum of cancer progression.