A Novel Diagnostic Assay for Oncogenic Human Papillomaviruses

DESCRIPTION (provided by applicant): Arbor Vita Corporation (AVC) has developed a novel cervical cancer diagnostic based on an understanding of the biology of human papillomavirus (HPV). HPV infection is one of the most common sexually transmitted diseases with an estimated 5.5 million new infections per year in the U.S. alone. High-risk (oncogenic) HPV types are correlated with virtually all cervical cancers. Pap smear and liquid based cytology screening has greatly reduced the incidence of cervical cancer, but the Pap test has both high false-negative and false-positive rates and requires an extensive infrastructure of trained cytologists to interpret the results. A cheaper test with greater reliability and predictive value would be of great clinical benefit. The virally-encoded E6 and E7 proteins of high-risk HPV types have been shown to be essential for cell transformation and cancer progression and E6 proteins from high-risk HPV types, but not low-risk HPV types, are known to bind cellular PDZ domains. AVC has extended those studies and demonstrated a perfect correlation between high-risk HPV and E6-PDZ binding. Based on these findings, we have developed a novel cervical cancer diagnostic assay of HPV E6 using PDZ protein capture. In our SBIR Phase I, AVC developed a novel PDZ capture sandwich ELISA test for HPV E6 that detects over 75% of high-risk HPV types and demonstrated its utility with human cancer samples. We were able to begin quantifying E6 from cells and improved sensitivity to allow E6 detection in a much smaller sample than typically collected in a Pap test. In Phase II, we propose to complete development of our prototype PDZ-based cervical cancer test in preparation for clinical trials. Specifically, we propose to: 1. Expand our antibody detection to include 95% of known high-risk HPV types. 2. Optimize clinical sample handling for E6 protein detection. 3. Optimize the PDZ/antibody sandwich ELISA for clinical laboratory implementation. 4. Extend the studies correlating high-risk E6 protein levels and clinical cytology staging.