Dr Roberts

Prof Woodman

We wish to understand how HPV proteins facilitate the normal infectious cycle, and the contribution they might make to the development of cervical neoplasia. Our functional studies are focussed on E4, an HPV protein that is abundant in infected cells, and on the HPV E6 oncoprotein. Perturbations of cellular pathways controlling cell cycle transitions, and disruption of nuclear ND10 domains are actions of E4 that may alter the host cell milieu to one that can facilitate HPV replication and production of progeny. We are now evaluating the contribution of E4 to the infectious cycle using in vitro models.

E6 interacts with cellular factors containing PDZ domains involved in controlling cell polarity and proliferation. We have shown that interaction with these PDZ proteins contributes to morphological transformation by E6. Expression of these proteins is changed during the progression of cervical disease. To study the natural history of HPV infection and its relationship to

cervical neoplasia we have recruited cohorts that provide sequential observations on the natural history of CIN. One of these cohorts, which included over 1000 women who were HPV negative and disease-free at time of study, revealed the temporal relationship between the acquisition of HPV infection, the first detection of cytological abnormality, and the risk of subsequent progression to high-grade CIN. We are now investigating the risk of progressive disease in relation to viral load, HPV-type, and viral integration.

Although methylated forms of tumour suppressor genes have been detected in cervical smears, it is unknown whether they are necessary or sufficient for the initiation or progression of disease. Using in vitro models of HPV immortalization, and CpG/gene expression arrays, we aim to provide an unbiased estimate of the epigenetic and transcriptional changes associated with HPV type-specific infection and viral integration.

The reorganization of an ND10 domain protein [PML, green] to intranuclear E4 [red] inclusions in a productive HPV1 infection. Arrowheads indicate basal cells

Altered expression of the E6 cellular target, discs large, during progression of cervical disease.

• Knight GL, et al (2004) Cooperation between different forms of the human papillomavirus type 1 E4 protein to block cell cycle progression and inhibit cell DNA synthesis. J Virol 78: 13920-33.
• Roberts S, et al (2003) The ND10 component promyelocytic leukaemia protein relocates to HPV type 1 E4 intranuclear inclusion bodies in cultured keratinocytes and in warts. J Virol 77: 673-
• Watson RA, et al (2003) Activity of the HPV E6 PDZ-binding motif correlates with enhanced morphological transformation in immortalized human keratinocytes. J Cell Sci. 116: 4925-34.
• Woodman CBJ, et al (2003) Human papillomavirus type 18 and rapidly progressing cervical intraepithelial neoplasia. Lancet 361: 40-43.