SPECIFIC INHIBITION OF HER2/NEU TRANSCRIPTION ELONGATION

描述：HER 2/neu癌基因似乎在肿瘤发生中起重要作用。 许多类型的人类癌症的发生和发展，包括 约25%的非小细胞肺癌（NSCLC）， 在美国，男性和女性的死亡原因。 该项目旨在寻找新的方法来特异性抑制HER 2/neu表达， 开发寡核苷酸（ODN）-苯丁酸氮芥（CHL）缀合物， 通过三链DNA形成以位点特异性方式与HER 2/neu基因结合 并导致CHL在特定鸟嘌呤碱基处的DNA烷基化。 具体而言，本申请中概述的工作将完成 以下目标：1）表征苯丁酸氮芥结合的能力 β（天然）和α（核酸酶抗性修饰）异头ODN， HER-2/neu基因中的直接位点特异性DNA烷基化; 2）表征 苯丁酸氮芥缀合的ODN结合HER/neu基因的能力， 抑制HER-2/neu基因转录延长体外和cDNA 表达质粒转染HeLa细胞和NIH 3 T3细胞; 3） 证明ODN结合和染色质中的位点特异性DNA修饰 表达HER-2/neu基因的活的NSCLC细胞; 4）表征 腺病毒介导ODN摄取和核定位的能力， 当用化学接头形成腺病毒-ODN复合物时， 5)确定最佳递送的ODN-CHL缀合物与人的免疫应答的能力。 抑制HER-2/neu基因表达，逆转肿瘤细胞恶性表型。 人NSCLC的组织培养和啮齿动物模型。 的具体目标 本申请旨在解决 成功开发基于ODN的位点特异性DNA结合药物。 的 这些具体目标的成功实现将带来宝贵的 深入了解基因特异性DNA结合化合物的设计。 的 HER-2/neu基因特异性抗基因化合物的开发将提供 关于HER-2/neu基因在肿瘤中的作用的大量信息， NSCLC的发生和进展，并可能导致新的治疗 用于HER-2/neu基因表达癌症如NSCLC的方法。

DESCRIPTION: The HER2/neu oncogene appears to play an important role in the initiation and progression of many types of human cancer, including approximately 25 percent of non-small cell lung cancer (NSCLC), the leading cause of death in both men and women in the U.S. The overall goal of this project is to find novel ways to specifically inhibit HER2/neu expression by developing oligonucleotide (ODN) - chlorambucil (CHL) conjugates that will bind in a site-specific manner to the HER2/neu gene by triplex DNA formation and lead to DNA alkylation at specific guanine bases by the CHL. Specifically, the work outlined in this application will accomplish the following goals: 1) Characterize the ability of chlorambucil-conjugated beta (natural) and alpha (nuclease resistant modification) anomeric ODNs to diret site-specific DNA alkylation in the HER-2/neu gene; 2) Characterize the ability of chlorambucil-conjugated ODNs to bind to the HER/neu gene and inhibit HER-2/neu gene transcription elongation in vitro and in a cDNA expression plasmid transfected into HeLa cells and NIH3T3 cells; 3) Demonstrate ODN binding and site-specific DNA modification in the chromatin of living NSCLC cells that express the HER-2/neu gene; 4) Characterize the ability of adenoviruses to mediate ODN uptake and nuclear localization in NSCLC cells when adenovirus-ODN complexes are formed with a chemical linker. 5) Determine the ability of optimally delivered ODN-CHL conjugates to inhibit HER-2/neu gene expression and reverse the malignant phenotype in tissue culture and rodent models of human NSCLC. The specific objectives of this application are designed to address the major obstacles to the successful development of an ODN-based site-specific DNA binding drug. The successful completion of these Specific Aims will lead to invaluable insights into the design of gene-specific DNA binding compounds. The development of a HER-2/neu gene specific anti-gene compound will provide a great deal of information about the role of the HER-2/neu gene in the initiation and progression of NSCLC and may lead to novel treatment approaches for HER-2/neu gene expressing cancers such as NSCLC.