Telomerase Transport and Targeting

端粒酶运输和靶向

• 批准号: 7259628
• 负责人: MICHAEL P TERNS
• 金额: $ 4.49万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7259628
• 关键词: DNA damage; RNA binding protein; RNA directed DNA polymerase; Xenopus oocyte; cell cycle; cell migration; crosslink; enzyme activity; enzyme complex; enzyme inhibitors; gel mobility shift assay; growth inhibitors; neoplastic process; polymerase chain reaction; ribozymes; telomerase

DESCRIPTION (provided by applicant): Human telomerase, the ribonucleoprotein enzyme that maintains telomeres at chromosome termini, is absent in most normal somatic cells and is present in nearly all cancer cells. Activation of telomerase induces cellular immortalization and is critical for the development and progression of tumors. Thus, inhibition of telomerase activity offers a promising approach for treating nearly all cancers. A better understanding of telomerase is needed to allow rational design of effective inhibitors. In humans, telomerase includes two subunits essential for function: an RNA subunit (human telomerase RNA) and a protein subunit (human telomerase reverse transcriptase). The location and mechanism of assembly of the enzyme complex in cancer cells (which may indicate where and how to target telomerase) is not known. Furthermore, identification of additional essential components of telomerase would provide additional potential targets for inhibition. The major objectives of this proposal are to obtain a detailed understanding of the trafficking and assembly of telomerase in cells, and to develop a class of RNA-based telomerase inhibitors effective at limiting or preventing the growth of cancer cells. In vivo analysis will be performed both in Xenopus oocytes (due to the many technical advantages of the system) and cultured human cells (including primary and cancer cell lines). To address our objectives we have defined the following three specific aims: Aim 1: To investigate the pathway of biogenesis of functional telomerase in vivo Aim 2: To examine the localization and trafficking of key human telomerase components in normal and cancer cells Aim 3: To develop efficacious anti-telomerase ribozymes capable of preventing growth of cancer cells.

描述（由申请人提供）：人端粒酶是一种将端粒维持在染色体末端的核糖核蛋白酶，在大多数正常体细胞中不存在，但在几乎所有癌细胞中都存在。端粒酶的激活可诱导细胞永生化，对于肿瘤的发生和进展至关重要。因此，抑制端粒酶活性为治疗几乎所有癌症提供了一种有前景的方法。需要更好地了解端粒酶，以便合理设计有效的抑制剂。在人类中，端粒酶包括两个对功能至关重要的亚基：RNA 亚基（人端粒酶 RNA）和蛋白质亚基（人端粒酶逆转录酶）。癌细胞中酶复合物组装的位置和机制（这可能表明端粒酶的靶向位置和方式）尚不清楚。此外，端粒酶其他必需成分的鉴定将为抑制提供额外的潜在靶标。该提案的主要目标是详细了解细胞中端粒酶的运输和组装，并开发一类基于 RNA 的端粒酶抑制剂，可有效限制或预防癌细胞的生长。将在非洲爪蟾卵母细胞（由于该系统的许多技术优势）和培养的人类细胞（包括原代细胞系和癌细胞系）中进行体内分析。为了实现我们的目标，我们定义了以下三个具体目标： 目标 1：研究功能性端粒酶体内生物发生的途径 目标 2：检查关键人类端粒酶成分在正常细胞和癌细胞中的定位和运输 目标 3：开发能够阻止癌细胞生长的有效抗端粒酶核酶。