FA-dependent HPV replication and transformation
Organotypic raft culture of HPV16 positive immortalized keratinocytes with immunofluorescence for cytokeratin 10 (green), a marker of the suprabasal layer, keratin 14 (red), a marker of the basal layer, and DAPI (blue), a marker of cell nuclei.
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Fanconi anemia (FA) is a genome instability syndrome that is characterized by progressive bone marrow failure and a high risk of cancer. Thirteen complementation groups and the corresponding FA genes have been identified. One of the main functions of FA protein products are to assemble into nuclear core complexes in response to DNA damage, and to subsequently mediate DNA repair. FA patients are particularly susceptible to leukemia as well as squamous cell carcinomas (SCCs) of the head and neck, anogenital region and skin. HPV is known to be associated with these types of SCCs in the general population. Previous data from our laboratory have shown that one of the HPV oncogenes, E7, can regulate the levels of Fanconi anemia proteins through activation of E2Fs. Additionally, we have used an organotypic epithelial raft culture system to show that patient keratinocytes that are deficient in FA develop exaggerated HPV-associated hyperplasia. We hypothesize that FA genes control proliferation, and that loss of FA genes thus promotes squamous cell carcinoma in HPV-positive, and perhaps HPV-negative, differentiated epidermis.
Research in the laboratory seeks to further elucidate the relationship between HPV and the Fanconi anemia DNA damage response pathway. We use a variety of molecular biology techniques, tissue culture, organotypic raft culture generation, and mouse xenograft experiments to accomplish this. Additionally, we are uniquely partnered with the Fanconi Anemia Comprehensive Care Center (FACCC) here at Cincinnati Children’s Hospital to acquire patient samples for basic science and epidemiological studies.
