Identifying Genes that Cause Lung Cancer to Progress

识别导致肺癌进展的基因

• 批准号: 6619197
• 负责人: ROBERT J DEBS
• 金额: $ 41.99万
• 依托单位: CALIFORNIA PACIFIC MED CTR RES INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6619197
• 关键词: biotechnology; gene delivery system; gene expression; genetic mapping; laboratory mouse; lung neoplasms; metastasis; microarray technology; neoplasm /cancer genetics; neoplasm /cancer transplantation; oncogenes; plasmids; ribozymes; transfection; transfection /expression vector; biotechnology; gene delivery system; gene expression; genetic mapping; laboratory mouse; lung neoplasms; metastasis; microarray technology; neoplasm /cancer genetics; neoplasm /cancer transplantation; oncogenes; plasmids; ribozymes; transfection; transfection /expression vector

DESCRIPTION (provided by applicant): Recently, cDNA microarray studies have identified multiple different genes that are aberrantly-expressed in aggressive human lung cancers. However, to translate these microarray-based results into improved therapies for cancer patients, the critical genes whose aberrant expression actually controls metastatic spread must now be identified. To date, functional assays for identifying such metastasis-controlling genes have been limited. Systemic gene transfer in adult, tumor-bearing mice is a powerful new approach to this problem. We now propose to use new systemic gene transfer technologies to identify which of nine selected genes (already identified by cDNA microarray studies to be aberrantly-expressed in poor prognosis, primary human lung cancers) actually control the metastatic spread of two different murine lung cancer tumor lines, LLC-LM and LLC-HM, in tumor-bearing mice. Our three specific aims are: Aim 1: To maximize the level and duration of gene expression produced in tumor-bearing mice by PEI- and IV, CLDC-based gene delivery. We will identify the most efficient, durably-expressing systemic gene transfer approaches for transfecting LLC-LM and LLC-HM cells in tumor-bearing mice. To accomplish this, we will optimize the: A) DNA carrier system, B) plasmid DNA:carrier ratio and C) repetitive dosing schedule. Aim 2: To use systemic delivery of selected cDNAs and ribozymes to identify specific genes that control the metastatic spread of lung cancers in mice. We will identify specific genes whose blocked expression or over-expression significantly reduces the metastatic spread of LLC-LM and HM in mice. Specifically, we will use plasmid-based ribozymes to target the endogenous expression of the CD98, cathepsin L, prostaglandin E synthase or VEGF-C genes, and use plasmid-based cDNAs to over-express the p21, 14-3-3, TGF-beta, thymosin beta or diacylglycerol genes. Each of these nine genes has already been identified by cDNA microarray studies to be aberrantly-expressed specifically in poor prognosis human lung cancers, and has been linked to the malignant phenotype. Aim 3: To identify pairs of metastasis-controlling cDNAs and/or ribozymes that act synergistically. We will assess IV co-injection of pairs of plasmid-based ribozymes and/or cDNAs that produce significant metastasis-controlling effects against murine lung cancer lines in Aim 2. We will assess multiple different dose combinations of each pair tested to determine whether there are specific interactions between each pair of genes. Overall, we will attempt to identify specific genes whose altered expression is functionally required for the metastatic spread of lung cancers. These critical metastasis-controlling genes can be specifically targeted by emerging small molecule-, protein- and gene-based therapeutic approaches; including those based on the more powerful systemic gene transfer approaches we have recently developed.

描述（由申请人提供）： 最近，cDNA微阵列研究发现了多种不同的基因，这些基因在侵略性的人类肺癌中异常表达。但是，为了将这些基于微阵列的结果转化为改进的癌症患者的疗法，现在必须确定其异常表达实际上控制转移性扩散的关键基因。迄今为止，用于鉴定这种转移控制基因的功能测定受到限制。成人的全身基因转移，含有肿瘤的小鼠是解决此问题的一种有力的新方法。现在，我们建议使用新的全身基因转移技术来确定哪种选定基因中的哪些基因（通过cDNA微阵列研究已经鉴定出在不良预后中异常表达的，原发性人类肺癌）实际上控制了两种不同的鼠肺癌tumor tumor系的转移性，LLC-LMM和LLC-HM，LLC-HM和LLC-HM，在Tumor-bearter的小鼠中。我们的三个具体目的是：目标1：通过PEI-和IV，基于CLDC的基因递送在肿瘤小鼠中产生的基因表达的水平和持续时间。我们将确定用于肿瘤小鼠中转染LLC-LM和LLC-HM细胞转染LLC-LM和LLC-HM细胞的最有效，表达最持久的全身基因转移方法。为此，我们将优化：a）DNA载体系统，b）质粒DNA：载体比和c）重复给药时间表。 AIM 2：使用选定的cDNA和核酶的全身递送来识别控制小鼠肺癌转移性扩散的特定基因。我们将确定其阻断表达或过表达的特定基因可显着降低小鼠中LLC-LM和HM的转移扩散。具体而言，我们将使用基于质粒的核酶来靶向CD98，组织蛋白酶L，Prostaglandin E合酶或VEGF-C基因的内源性表达，并使用基于质粒的cDNA过表达P21，14-3-3，TGF-BETA，TGF-BETA，TGF-BETA，胸腺素beta beta beta beta oriacycylglycerol Genes。 cDNA微阵列研究已经鉴定出这9个基因中的每一个，它们在预后不良的人类肺癌中被异常表达，并且与恶性表型有关。目标3：识别成对的转移控制的cDNA和/或核酶可以协同作用。我们将评估静脉注射的基于质粒的基于质粒的核酶和/或cDNA，这些核酶和/或cDNA对AIM 2中的鼠肺癌线产生明显的转移控制作用。我们将评估经过测试的每对测试的多个不同剂量组合，以确定每对基因之间是否存在特定的相互作用。总体而言，我们将尝试确定其在功能上需要改变表达的特定基因，即肺癌的转移扩散。这些关键的转移控制基因可以通过新兴的小分子，蛋白质和基因的治疗方法来特异性靶向。包括基于我们最近开发的更强大的系统基因转移方法的方法。