The overall objective of this project is to develop diagnostic assays that are capable of identifying a subset of patients with cervical dysplasia who are at increased risk of developing cervical cancer. Human papillomavirus (HPV) type 16 E6 or E7 oncoproteins are major causative agents in cervical cancer, and are expressed in high grade dysplastic lesions of the cervix and in cervical tumor tissue. The applicant's goal is to develop and evaluate assays of E6 and E7 expression that can be used to examine tissues from patients at risk for developing cervical cancer. He will use this information to determine whether E6 and E7 oncogene expression is associated with the grade of cervical dysplasia. At present the DNA that codes for HPV oncoproteins can be detected for diagnosis of high-risk HPV infection, but the presence of HPV type-specific DNA does not provide information about whether the oncoproteins are actually being expressed. Data from the applicant's preliminary studies using RT-PCR and RNA in situ hybridization demonstrated that E6 and E7 expression can be detected in the tissue of individuals with cervical dysplasia and cervical cancer. He hypothesizes that HPV 16 E6 and E7 oncoprotein expression, assessed using diagnostic assays of HPV oncogenes, oncoproteins or their antibodies in blood and tissues of patients with cervical dysplasia or cervical cancer will correlate with the grade of cervical dysplasia or cervical cancer. To test this hypothesis, he proposes to produce and E6 fusion protein, to develop new diagnostic assays for detecting E6 and E7 RNA and oncogene expression in cervical tissue, and to use the E6 fusion protein to develop assays for detecting antibodies specific against E6 and E7 oncoproteins in sera from patients at risk for developing cervical cancer. The assays of expression of these oncogenes and their antibodies can be used to monitor for the effectiveness of cervical cancer treatment modalities, and would allow investigators to make interventions in patients with intraepithelial neoplasia to decrease the risk of future development of invasive cancer.
| Wang-Johanning, Feng; Lu, Danielle W; Wang, Yueying et al. (2002) Quantitation of human papillomavirus 16 E6 and E7 DNA and RNA in residual material from ThinPrep Papanicolaou tests using real-time polymerase chain reaction analysis. Cancer 94:2199-210 |
