TRIPLEX BASED GENE THERAPY OF BREAST AND OVARIAN CANCER

乳腺癌和卵巢癌的三重基因治疗

• 批准号: 2769850
• 负责人: SCOT W EBBINGHAUS
• 金额: $ 7.88万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2769850
• 关键词: Adenoviridae; athymic mouse; breast neoplasms; disease /disorder model; gene expression; gene therapy; genetic promoter element; human genetic material tag; neoplasm /cancer genetics; neoplastic cell; oligonucleotides; oncogenes; ovary neoplasms; phenotype; recombinant virus; synthetic nucleic acid; tissue /cell culture; transcription factor; triple helix

DESCRIPTION (Applicant's Description): Dr. Ebbinghaus is a senior fellow who has devoted more than 60% of his last five years to basic research while completing clinical training in Internal Medicine, Medical Oncology, and Hematology at UAB. In addition to completing a formal core graduate program in Cell and Molecular Biology, Dr. Ebbinghaus joined an ongoing laboratory effort in Dr. Donald Miller's laboratory and initiated a program of designing triplex-based anti-gene strategies to specifically inhibit the HER-2/neu oncogene. Dr. Ebbinghaus will join the faculty of the Division of Hematology-Oncology at the completion of his fellowship and devote over 80% of his time to his laboratory efforts. Dr. Ebbinghaus will be given laboratory space in close physical proximity to Dr. Miller's laboratory in the Wallace Tumor Institute so that he may continue to use the critical equipment in Dr. Miller's laboratory and the Animal Core Facility in this building. The NCI Clinical Investigator Award will support Dr. Ebbinghaus in his transition from a senior fellow to an independent investigator in his own laboratory. The HER-2/neu oncogene plays an important role in many types of human cancer. There is good data to support the concept that inhibiting HER-2/neu expression will reverse the malignant phenotype and have beneficial clinical effects in patients with HER-2/neu related malignancies. Oligonucleotides are particularly attractive as specific inhibitors of oncogene expression since they have the ability to bind to target DNA or RNA sequences with very high specificity. Dr. Ebbinghaus and Dr. Miller have investigated the ability of DNA oligonucleotides to bind to the HER-2/neu oncogene promoter and found that these oligonucleotides potently and specifically inhibit HER-2/neu transcription through DNA triplex formation in vitro. The broad goals of this proposal are to determine the ability of triplex formation to inhibit HER-2/neu expression in cell culture and animals by both synthetic DNA oligonucleotides and RNA oligonucleotides transcribed in vivo. This proposal will build on Dr. Ebbinghaus and Dr. Miller's previous experience with triplex forming oligonucleotides in vitro and current work using gene therapy vectors to deliver these oligonucleotides to tumor cells in culture. The specific aims of this proposal are: 1) to characterize the ability of synthetic DNA oligonucleotides to inhibit HER-2/neu expression and alter the malignant phenotype of HER-2/neu expressing tumor cell lines in culture; 2) to characterize the ability of intracellularly transcribed RNA oligonucleotides to form triplex structures in the HER-2/neu promoter and inhibit gene expression in tumor cells in culture; 3) to generate a recombinant adenovirus capable of expressing RNA oligonucleotides designed form triplex structures in the HER-2/neu promoter and characterize its ability to inhibit HER-2/neu expression in tumor cells; 4) to develop an animal model to test the efficacy of synthetic DNA, transcribed RNA, and recombinant adenovirus expressed oligonucleotides to inhibit HER-2/neu gene expression in vivo. The successful completion of these specific aims will generate invaluable insights into the role of HER-2/neu in the cause of certain cancers as well as provide the background for triplex based gene therapies for HER-2/neu related cancer.

描述（申请人的描述）：艾宾浩斯博士是一名高级研究员 他在过去五年中将60%以上的时间用于基础研究， 完成内科、肿瘤内科的临床培训， 血液学在UAB。 除了完成正式的核心研究生课程外， 在细胞和分子生物学方面，艾宾浩斯博士加入了一个正在进行的实验室， 唐纳德米勒博士的实验室的努力，并启动了一个计划， 设计基于三链体的抗基因策略， HER-2/neu癌基因。 艾宾浩斯博士将加入该部门的教师， 他在完成奖学金后，将80%以上的时间用于血液肿瘤学， 他的实验室工作的时间。 艾宾浩斯博士将获得 实验室空间在物理上接近米勒博士的实验室， 华莱士肿瘤研究所，以便他可以继续使用关键的 设备在米勒博士的实验室和动物核心设施在这一点上， 建设 NCI临床研究者奖将支持Ebbinghaus博士 在他从高级研究员转变为独立调查员的过程中， 自己的实验室。 HER-2/neu癌基因在许多类型的人类肿瘤中起重要作用， 癌 有很好的数据支持抑制HER-2/neu 表达将逆转恶性表型，并具有有益的临床意义。 HER-2/neu相关恶性肿瘤患者的疗效。 寡核苷酸 作为癌基因表达的特异性抑制剂特别有吸引力 因为它们具有与靶DNA或RNA序列结合的能力， 特异性高。 艾宾浩斯博士和米勒博士研究了 DNA寡核苷酸结合HER-2/neu癌基因启动子的能力 并发现这些寡核苷酸可以有效特异地抑制 HER-2/neu通过DNA三链体形成的体外转录。 广大 该建议的目的是确定三链体形成的能力， 抑制HER-2/neu在细胞培养物和动物中表达 体内转录的DNA寡核苷酸和RNA寡核苷酸。 这 一项建议将建立在艾宾浩斯博士和米勒博士以前的经验基础上 与体外形成三链体的寡核苷酸和目前的工作， 将这些寡核苷酸递送至培养的肿瘤细胞。 本建议的具体目标是：1）描述 合成的DNA寡核苷酸，以抑制HER-2/neu表达并改变HER-2/neu表达。 培养物中表达HER-2/neu的肿瘤细胞系的恶性表型; 2） 来表征细胞内转录的RNA 寡核苷酸以在HER-2/neu启动子中形成三链体结构， 抑制培养的肿瘤细胞中的基因表达; 3）产生 能够表达设计的RNA寡核苷酸的重组腺病毒 在HER-2/neu启动子中形成三链体结构，并表征其 抑制HER-2/neu在肿瘤细胞中表达的能力; 4）开发一种新的抗肿瘤药物， 动物模型来测试合成DNA、转录RNA和 重组腺病毒表达的HER-2/neu基因抑制寡核苷酸 体内表达。 这些具体目标的成功实现将 对HER-2/neu在以下原因中的作用产生宝贵的见解： 某些癌症以及提供了基于三链体的基因的背景 HER-2/neu相关癌症的治疗。