Molecular Dissection of Lung Cancer Progression and Metastasis

描述(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。肺癌患者预后不佳的原因有很多，但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，转移是一个主要的临床挑战，其驱动因素是目前尚不清楚的细胞状态改变。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背后的分子和细胞变化。我们将使用一个基因工程的肺腺癌小鼠模型，它概括了人类肺腺癌的遗传变化和组织学进展。在Aim1中，我们将通过慢病毒载体系统在体内直接表达肺癌转移的合理候选调控因子，从而在体内功能上询问肺癌转移促进和抑制基因。我们将把这些工具与肺腺癌的基因工程小鼠模型结合起来，该模型结合了癌细胞的荧光标记，从而能够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将肺腺癌细胞从原发肿瘤和转移瘤中分离出来，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 揭示了原发肿瘤的整体基因表达与不同器官转移的关系。在AIM3中，我们将通过人类肺腺癌候选基因表达与临床结果的相关性，以及使用明确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，来分析和功能剖析人类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了解这种疾病状态的分子基础方面的差距，临床实践和患者结果都将受到任何新疗法的极大影响，这些新疗法可能来自我们拟议的分析所获得的基本知识。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理，并最终发现跨越癌症进展连续体的新治疗靶点。

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.