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.
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.
