Molecular Dissection of an Arntl2 induced pro-metastatic secretome

Arntl2 诱导的促转移分泌蛋白组的分子解剖

基本信息

批准号：
9260764
负责人：
Monte Meier Winslow
金额：
$ 36.7万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-12 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9260764
关键词：
Adhesions Benign Biological Biological Assay CRISPR/Cas technology Cancer Etiology Cancer Patient Cell Culture System Cell Culture Techniques Cell Death Cell Line Cells Cessation of life Clinical Complex Country Data Development Disease Dissection Disseminated Malignant Neoplasm Distant Environment Epithelial Cells Gene Expression Profiling Genes Genetic Genetic Engineering Genetically Engineered Mouse Goals Growth Health Human In Vitro Individual Knowledge Lung Adenocarcinoma Malignant Neoplasms Malignant neoplasm of lung Mediating Metastatic Adenocarcinoma Methods Modeling Molecular Mus Neoplasm Metastasis Organ Outcome Output Pathway interactions Patient-Focused Outcomes Patients Phenotype Positioning Attribute Primary Neoplasm Process Proteins Recombinant Proteins Role Sampling Signal Transduction Site Solid Neoplasm System Testing Transcriptional Regulation Transplantation United States Woman autocrine cDNA Expression cancer cell cancer type clinical practice experimental study fitness genome editing in vivo knock-down lentiviral-mediated loss of function men metastatic process mouse model neoplastic cell new therapeutic target novel novel therapeutics overexpression prevent public health relevance response screening therapeutic target transcription factor treatment strategy tumor tumor progression tumorigenesis

项目摘要

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. By integratin gene expression analyses on murine models of metastatic lung adenocarcinoma and human lung adenocarcinomas we identified the transcription factor Arntl2 as a key driver of lung adenocarcinoma metastatic ability. Arntl2 appears to drive metastatic fitness by controlling the expression of a complex pro-metastatic secretome that has the ability to greatly increase clonal growth potential. In Aim1, we will functionally interrogate Arntl2 function in human and mouse lung adenocarcinoma cell lines. We will perform gain- and loss-of-function experiments and fully assess the cellular phenotypes driven by Arntl2. Transplantation assays and quantification of initial adhesion, proliferation, and cell death within the metastatic site in vivo will elucidate te cellular consequence of high Arntl2 expression. In Aim2, we will investigate which Arntl2-regulated secreted factors cooperate to drive clonal growth and metastatic ability. We will integrate screening of recombinant proteins in cell culture, gain- and loss-of-function experiments in cell culture and in vivo, and therapeutically target pathways downstream of key pro-metastatic secreted proteins to better understand the importance of autocrine metastatic niche factors. In Aim3, we will use novel methods for CRISPR/Cas9-mediated genome editing and lentiviral-mediated cDNA expression to test the requirement and sufficiency of Arntl2 and Arntl2- regulated genes to promote step of the metastatic cascade in autochthonous mouse models of human lung cancer. 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 reveal novel therapeutic targets across the continuum of cancer progression.

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，仅在美国每年就有超过 155,000 名患者死亡，有几个因素导致肺癌患者的预后不良。但是，与大多数实体瘤一样，癌细胞离开原发肿瘤并建立无法手术转移的能力是成功治疗的主要障碍，因此转移代表了目前尚不完善的临床挑战。通过对转移性肺腺癌和人肺腺癌的小鼠模型进行整合素基因表达分析，我们确定转录因子 Arntl2 是肺腺癌转移能力的关键驱动因素，Arntl2 似乎通过控制复合物的表达来驱动转移适应性。能够大大增加克隆生长潜力的促转移分泌蛋白组在 Aim1 中，我们将在功能上探讨 Arntl2 在人和小鼠肺中的功能。我们将进行功能获得和功能丧失实验，并全面评估 Arntl2 驱动的细胞表型，并对体内转移部位的初始粘附、增殖和细胞死亡进行量化，以阐明细胞结果。在 Aim2 中，我们将研究哪些 Arntl2 调节的分泌因子协同驱动克隆生长和转移能力。细胞培养、细胞培养和体内功能获得和功能丧失实验以及关键促转移分泌蛋白下游的治疗靶标途径，以更好地了解自分泌转移生态位因子的重要性在 Aim3 中，我们将使用新方法。用于 CRISPR/Cas9 介导的基因组编辑和慢病毒介导的 cDNA 表达，以测试 Arntl2 和 Arntl2 调节基因促进转移级联步骤的必要性和充分性考虑到转移性癌症的巨大临床影响以及目前对这种疾病状态分子基础了解的差距，临床实践和患者结果都将受到任何可能源自基本治疗的新疗法的极大影响。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，并最终揭示整个癌症进展的新治疗靶点。