The primary objective of this proposal is to study women with multicentric neoplastic disease of the anogenital epithelium to compare the pathobiology of human papillomavirus (HPV) infection at different epithelial surfaces in the same person. A selected number of ovarian and endometrial lesions will also be examined to assess the potential susceptibility to HPV infection of other reproductive/genital epithelia.
The specific aims of this proposal are, in summary: 1) To identify and characterize HPV genomes found in each lesion in women with multifocal neoplasias, and to relate the transcription and expression of HPV genes and proteins to the histopathologic and clinical aspects of disease, 2) To investigate the effects of HPV infection of squamous cells at different epithelial surfaces by comparing the expression of growth factor genes, growth factor receptor proteins and cell differentiation-specific proteins in multicentric lesions, and 3) To examine lesions of other reproductive/genital epithelia for the presence of HPV nucleic acid sequences and the expression of HPV genes and proteins. This study will use existing formalin-fixed and paraffin-embedded tissue specimens for in situ hybridization analyses. HPV DNA genomes will be identified and localized using biotin-labelled HPV DNA probes and 3H-labelled HPV RNA probes. In a small number of prospectively identified patients, HPV genomes found at each site in multicentric disease will be examined in detail by restriction enzyme digestion and Southern transfer hybridizations. Transcription of HPV and growth factor genes will be studied by in situ hybridization with 3H-labelled probes. HPV RNA probes will be generated from defined open reading frames and exon-specific probes will also be used. Growth factor gene expression will be studied with cDNA probes. The expression of HPV-specific, cell differentiation- specific and growth factor receptor proteins will be examined with immunohistochemical techniques.
