Arbor Vita proposes to validate the interactions of oncogenic E6 variants with a specific cellular protein family as a target for therapeutic intervention in Human papilloma virus (HPV) infection and cervical cancer. Oncogenic variants of HPVs have been correlated with cervical cancer at a frequency of 99.7%. Published and unpublished data has correlated HPV oncogenicity with variants of HPV E6 oncoprotein that contain a particular binding motif or ligand. Specifically, Arbor Vita proposes to 1) identify the human protein family members targeted by oncogenic E6 variants 2) confirm expression of target proteins in cervical cell lines 3) confirm target interaction occurs in vivo 4) demonstrate that interfering with the oncogenic E6 ligand/cellular protein interaction reduces oncogenic transformation in cellular assays and, 5) develop an in vitro screen for small molecule inhibitors of the oncogenic E6 ligand/protein interaction. To accomplish these aims Arbor Vita will utilize it's proprietary in vitro assay that can be used to screen interactions, determine affinity, and screen small molecules for the ability to disrupt the targeted interaction. Included are peptide production/purification, cloning, protein production/purification, and small molecule synthesis. Cell based assays including soft agar/transformation assays and qPCR analysis will be used to determine in vivo effects. ? ? ?
