High Dimensional Analyses of the Mucosal Microenvironment in Precancerous, Intraepithelial Cervical HPV Disease
Cornelia L. Trimble
Johns Hopkins University School of Medicine, Baltimore, MD, USA
Malignancies caused by human papillomavirus, including cancers of the cervix, vagina, vulva, anus, and oropharynx, comprise 1/3 of pathogen-associated cancers. In the cervix, intraepithelial precancerous lesions (CIN2/3) is associated with integration of the viral genome, and subsequent constitutive expression of two viral oncoproteins, E6 and E7, in a functionally obligate manner. However, CIN2/3 is clinically indolent, and can even undergo ‘spontaneous’ regression. Because these lesions are directly accessible, this clinical setting provides an opportunity to assess tissue before and after study interventions, and thereby assess proof-of-principle in immunotherapeutic strategies. Our analytic approach begins with quantitative multiplex digital image analysis to determine the intensity and colocalization of cells expressing antigens of interest.These data provide a histologic context for analyses of subsequent rapid immuno-laser capture microdissection of specific cell subsets in subject-matched tissue blocks. Molecular studies can then be performed on these relatively clean samples. Using this approach, we have identified mechanisms of immune evasion that we are now manipulating directly in the mucosa, in combination with therapeutic vaccination targeting HPV16 and 18 E6 and E7, prior to standard therapeutic resection.
Funding: NIH R01 CA142691, P50 CA098252, 1R21 CA123876, 5P30 CA6973, Su2C/Farrah Fawcett Foundation, the Dana Foundation, the Commonwealth Foundation.