ART On Oral Tissue Growth, Function, and HPV Infections

描述(申请人提供)：父母R01的目标是调查抗逆转录病毒治疗(ART)对不同类型口腔上皮中HPV感染的生命周期的影响。我们的假设是ART影响宿主组织和/或HPV的自然历史，增加HPV在口腔上皮组织中感染和诱导病理的能力。我们推测，抗逆转录病毒药物影响口腔组织的方式导致不利的口腔健康。我们假设的基本原理是基于HIV患者在开始高效抗逆转录病毒治疗(HAART)后口腔HPV病变的频率增加，并且所涉及的HPV基因型通常与相同的解剖区域不相关。目标和理由没有改变。到目前为止，我们积累的数据支持了我们的假设和追求父母申请目标的理论基础。母体应用有三个目的(1)确定抗逆转录病毒(ART)药物对三维口腔上皮组织的影响；(2)ART对口腔上皮中HPV允许性复制和感染的影响；(3)在ART治疗的背景下研究口腔上皮中HPV的完整生命周期。对于这一竞争性修订申请，我们决定将重点放在一个实验设计上，该设计将提供公正的分析，并在分配的时间内最有能力扩大父提案的范围。NICDR/NIH认识到，关于ART对口腔粘膜的影响以及ART对口腔组织HPV感染的影响，人们知之甚少。我们已经完成的研究表明，抗逆转录病毒药物可以对牙龈和扁桃体上皮组织的生长和分化表型产生显著影响。现在，我们希望通过进行微阵列分析，将这些研究提升到一个新的水平，以确定用抗逆转录病毒药物治疗口腔组织所诱导的基因表达的全球变化。我们期望微阵列分析能够公正地测量扁桃体和上皮组织中受常用抗逆转录病毒药物影响的基因表达的变化。我们预计这将揭示与生物学功能相关的途径和基因集，例如细胞生长、癌变、伤口愈合和分化等，这些途径和基因集与我们观察到的表型变化和患者观察到的不良副作用有关。我们还成功地培养出了持续感染HPV的扁桃体和牙龈细胞系。我们正在提议进行微阵列研究，以衡量HPV16和HPV32对口腔组织的广泛影响，并提供数据，使我们能够比较和对比不同病毒对不同组织的影响。这将使我们能够比较口腔和宫颈组织中HPV16感染引起的差异和相似之处，从而深入了解病毒特异性和组织特异性的肿瘤发生机制。目前对微阵列技术的投资将提供与母公司提案范围的文字和精神一致的公正分析，大大扩大了提案的范围。 公共卫生相关性：到目前为止收集的数据证实了我们的假设，即抗逆转录病毒药物以一种导致不利口腔健康的方式影响口腔组织。我们建议进行微阵列研究，以衡量HPV16和HPV32对口腔组织的广泛影响。这将提供不偏不倚的分析，显著扩大提案的范围。

DESCRIPTION (provided by applicant): The goal for the parent R01 was to investigate the effect of antiretroviral therapy (ART) on the life cycle of oral HPV infections in different types oral epithelium. Our hypothesis was that ART affects the host tissue and/or the natural history of HPV, increasing HPV's ability to infect and induce pathology in oral epithelial tissue. We speculated that ART drugs affect oral tissue in a manner that leads to adverse oral health. The rationale for our hypothesis was based on the increased frequency of oral HPV lesions in HIV patients after they begin highly active antiretroviral therapy (HAART) and that the HPV genotypes involved are not normally associated with the same anatomical areas. The goals and rationale have not changed. The data we have accumulated to this point has strengthened our hypothesis and rationale for pursuing the aims of the parent application. The parent application had three aims (1) Determine the effects of anti-retroviral (ART) drugs on three-dimensional oral epithelial tissues; (2) The effect of ART on HPV permissive replication and infection in oral epithelium; and (3) Study the complete HPV life cycle in oral epithelium in the context of ART treatment. For this Competitive Revision application we decided to focus on an experimental design that will provide an unbiased analysis with the greatest ability to expand the scope of the parent proposal within the time resources allotted. The NICDR/NIH recognized that very little is known concerning the effects of ART on oral mucosa and by extension the effects of ART on HPV infection of oral tissues. We have completed studies demonstrating that the ART drugs can have dramatic effect on the growth and differentiation phenotype of gingival and tonsil epithelial tissues. We now want to take these studies to the next level by performing microarray analyses to identify the global changes in gene expression induced by treating oral tissues with the ART drugs. We expect the microarray analyses to provide an unbiased measurement of changes of gene expression in tonsil and epithelial tissues, affected by commonly used ART drugs. We expect that this will reveal pathways and gene sets relating to biological functions such as cell growth, carcinogenesis, wound healing, and differentiation to name a few that would relate to the changes in phenotype we have observed and to the adverse side effects observed in patients. We have also been successful in developing continuously infected HPV tonsil and gingival cell lines. We are proposing microarray studies to measure the wide spread impact that HPV16 and HPV32 have on oral tissue and provide data that will allow us to compare and contrast the impact of different viruses on different tissues. This will allow us to compare the differences and similarities induced by HPV16 infection of oral and cervical tissues providing insights into virus-specific and tissue-specific mechanisms tumorigenesis. Investment at this time in microarray technology will provide unbiased analyses in line with the letter and spirit of the scope of the parent proposal, providing a significant expansion of the proposal's scope. PUBLIC HEALTH RELEVANCE: Our hypothesis that ART drugs affect oral tissue in a manner that leads to adverse oral health has been strengthened by data collected to this point. We propose to conduct microarray studies to measure the wide spread impact that HPV16 and HPV32 have on oral tissue. This will provide unbiased analyses, providing a significant expansion of the proposal's scope.