Effects of Antiretroviral Therapy on Telomerase Function in Human Oral Epithelium

抗逆转录病毒治疗对人口腔上皮端粒酶功能的影响

• 批准号: 7352682
• 负责人: Mo K. Kang
• 金额: $ 35.14万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7352682
• 关键词: 2' -Deoxythymidine; Acquired Immunodeficiency Syndrome; Adverse effects; Atrophic; Biological Process; Catalytic Domain; Cell Aging; Cell Proliferation; Cell division; Cells; DNA; DNA Repair; DNA lesion; Development; Double Strand Break Repair; Epithelial; Epithelial Cells; Erythema Multiforme; Exhibits; Family; Frequencies; Genetic; Genome; Goals; HIV; Hairy Leukoplakia; Highly Active Antiretroviral Therapy; Human; In Vitro; Individual; Laboratories; Laboratory Finding; Lead; Length; Maintenance; Mediating; Molecular; Mutation; Oral; Oral Manifestations; Oral candidiasis; Oral cavity; Oral mucous membrane structure; Outcome; Patients; Population; Prevention; Proliferating; Purpose; RNA; RNA-Directed DNA Polymerase; Recurrence; Reporting; Research Personnel; Retroviridae; Reverse Transcriptase Inhibitors; Role; Somatic Cell; Stem cells; Stratification; Telomerase; Telomerase RNA Component; Telomerase inhibition; Telomere Shortening; Testing; Ulcer; Virus Diseases; anti-cancer therapeutic; antiretroviral therapy; cancer cell; enzyme activity; human TERT protein; in vivo; keratinocyte; member; monolayer; novel; oral cavity epithelium; oral fibroblast; oral wart; prevent; programs; senescence; telomere

DESCRIPTION (provided by applicant): The long-term goal of this project is to reduce or reverse the oral complications of highly active antiretroviral therapy (HAART) in patients infected with the human immunodeficiency virus (HIV). Although the oral manifestations of HIV infection have significantly decreased after the introduction of HAART, several adverse effects have also been reported in the oral mucosa, including recurrent oral ulceration, epithelial atrophy, erythema multiforme, epithelial desquamation, eruptive chielitis, and multiple oral warts. The purpose of the current application is to elucidate the effects of telomerase inhibition by reverse transcriptase inhibitors (RTIs) in mediating the side effects of HAART. Telomerase is a cellular reverse transcriptase with at least two distinct biological functions in (1) synthesis of telomere DNA and (2) maintenance of genome integrity. Our laboratory found remarkably high level of telomerase activity in normal human oral keratinocytes (NHOK) derived from oral epithelium. Telomerase activity in NHOK is specifically associated with actively proliferating cells and is completely lost during cellular senescence. Importantly, telomerase activity can be effectively inhibited by several RTIs commonly used as HAART. The central hypothesis of this proposal is that telomerase inhibition in NHOK by RTIs is responsible for the diminution of regenerative capacity of the oral epithelium and adverse oral mucosal complications associated with long-term administration of HAART in HIV+ patients. To test our hypothesis, we propose three Specific Aims: (1) to determine the telomerase activity, telomeric status, and cellular phenotypic alterations in NHOK exposed to RTIs in vitro and in cells derived from HIV+ patients with and without HAART; (2) to determine the effects of RTI/HAART on the DNA repair activities, mutation frequency, and genetic integrity in NHOK; and (3) to investigate the effects of AZT on phenotypic alterations in NHOK expressing exogenous telomerase or acquiring enhanced replication potential. The aims 1 and 2 will investigate the detailed phenotypic and genetic effects of RTI/HAART in oral epithelium. In aim 3, we will determine whether augmenting cellular telomerase activity and/or "priming" the cells with enhanced replicative potential can prevent the adverse phenotypic effects of AZT. The outcome of this project will be used for prevention and management of oral manifestations of HIV infection and the acquired immunodeficiency syndrome (AIDS) by reducing the negative effects of HAART.

描述（由申请人提供）：该项目的长期目标是减少或逆转感染人类免疫缺陷病毒（HIV）患者的高效抗逆转录病毒疗法（HAART）的口腔并发症。虽然引进HAART后HIV感染者的口腔表现明显减少，但口腔粘膜也出现了一些不良反应，包括复发性口腔溃疡、上皮萎缩、多形性红斑、上皮剥落、发疹性唇炎和多发性口腔疣。本申请的目的是阐明逆转录酶抑制剂(RTI)抑制端粒酶在介导HAART副作用中的作用。端粒酶是一种细胞逆转录酶，具有至少两种不同的生物学功能：(1) 端粒 DNA 的合成和 (2) 维持基因组完整性。我们的实验室在源自口腔上皮的正常人口腔角质形成细胞（NHOK）中发现了非常高水平的端粒酶活性。 NHOK 中的端粒酶活性与活跃增殖的细胞特别相关，并且在细胞衰老过程中完全丧失。重要的是，端粒酶活性可以被常用的几种 RTI 有效抑制，如 HAART。该提议的中心假设是，RTIs 抑制 NHOK 中的端粒酶导致口腔上皮细胞再生能力下降，以及与 HIV+ 患者长期服用 HAART 相关的不良口腔粘膜并发症。为了检验我们的假设，我们提出了三个具体目标：(1) 确定体外暴露于 RTI 的 NHOK 以及来自接受或未接受 HAART 的 HIV+ 患者的细胞中的端粒酶活性、端粒状态和细胞表型改变； (2) 确定RTI/HAART对NHOK DNA修复活性、突变频率和遗传完整性的影响； (3) 研究AZT对表达外源端粒酶的NHOK表型改变或获得增强的复制潜力的影响。目标 1 和 2 将研究 RTI/HAART 对口腔上皮的详细表型和遗传影响。在目标 3 中，我们将确定增强细胞端粒酶活性和/或“启动”具有增强复制潜力的细胞是否可以预防 AZT 的不良表型效应。该项目的成果将用于通过减少HAART的负面影响来预防和管理艾滋病毒感染和获得性免疫缺陷综合症（艾滋病）的口腔表现。