HPV DIFFERENTIATION DEPENDENT REPLICATION

Project: Research project

Project Details

Description

Subsets of human papillomaviruses (HPV) are associated with malignant and
benign tumors of the anogenital region. Using a cell line derived from a
biopsy of a cervical intraepithelial neoplasia type 1 (CIN 1), we recently
demonstrated in an organotypic (raft) culture system the complete HPV
replication cycle concomitant with the induction of a more complete
differentiation program in vitro The use of our raft culture system with
its ability to reproduce the complete HPV replication cycle allows for a
complete description of HPV activity, transcriptional and translational,
in human epithelial tissue actively propagating virus. Our working
hypothesis is that HPV's replication cycle is intimately connected to the
growth and terminal differentiation program of epithelial tissues. Changes
that occur in epithelial cells as they undergo the process of
differentiation directly control the viral replication cycle at the
transcriptional and translational levels. To this end, we plan to define
viral transcriptional and translational activity and correlate this
activity to the temporal and spatial development of the natural host
tissue, human squamous epithelium. In addition, the second aspect of our
working hypothesis is that HPV infection and subsequent viral gene
expression alters the differentiation program of the host tissue. In the
case of the high risk HPV types, it is this interaction that results in
the eventual progression of the neoplasia to malignancy.

The first specific aim focuses on the patterns of HPV transcription in
differentiating human epithelium during the complete viral replication
cycle. The second specific aim focuses on HPV 31b protein expression
patterns in differentiating human epithelium. A comparison of viral
transcriptional and translational expression patterns will be made in (i)
differentiating cervical epithelium that has been induced by 12-O-
tetradecanoyl phorbol 13-acetate (TPA) to propagate HPV virions, (ii)
differentiating cervical epithelium not treated with TPA but latently
infected with HPV, and (iii) monolayer cultures of cervical cells. Using
in situ technologies, we will map the spatial patterns of the HPV 31b gene
expression in the context of the three-dimensional architecture in
stratified and differentiated human epithelium. The use of the raft
cultures will allow us to define the patterns of HPV gene expression
during the temporal development of the host tissue. The expression of
terminal epithelial differentiation markers including keratin 10,
filaggrin, and involucrin will be examined and compared with HPV gene
expression. The studies proposed in this application will enable us to
test our working hypothesis that HPV gene expression and replication is
contingent upon changes in the cellular environment of the epithelial cell
as it stratifies and differentiates.
StatusFinished
Effective start/end date3/1/962/29/00

Funding

  • National Cancer Institute
  • National Cancer Institute
  • National Cancer Institute

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