New therapeutics directed at treating pre-cervical cancer changes prior to the development of cervical cancer require staging of the molecular changes associated with transformation and carcinogenesis in order to treat at the earliest possible stage. To that end, we report the preliminary results of a study that uses a single, high throughput assay (Oncotect 3Dx) that defines the stages of squamous cell abnormalities that lead to cervical cancer.
We analyzed 227 samples that included 79 normals (NILM HRHPV DNA-), 72 low grade (NILM/ASCUS/LSIL HRHPV DNA+), and 76 high grade (HSIL HRHPV DNA+) collected in ThinPrep® liquid-based cytology media. Each sample was assayed using the 96-well OncoTect 3DX assay that quantifies E6, E7 mRNA and cell cycle on a cell by cell basis. In particular, the post-G0/G1% was calculated for each sample as a measure of cell proliferation. In addition, mean corpuscular volume (MCV) was determined for every cell in all samples.
There was an inverse correlation between cervical abnormality stage, normal-low grade-high grade, and MCV with normal samples being 161 uM3, low-grade 131 uM3, and high grade 113 uM3 (Mann-Whitney P=<0.001). The post-G0/G1% also differed depending on the stage of abnormality with normal samples and high grade samples having the highest proliferation rate and low grade abnormalities having the lowest proliferation rate (Mann-Whitney P=0.03).
Using multiple parameters quantified using the OncoTect 3Dx assay, we were able to define normal cervical samples as E6, E7 mRNA-, MCV hi, post-G0/G1% hi; low grade cervical samples as E6, E7 mRNA +/-, MCV intermediate, post-G0/G1% low; and high grade cervical samples as E6, E7 mRNA +, MCV low, post-G0/G1% high. The ability to stratify cervical cancer abnormalities in an automated, high-throughput manner is advantageous for companion diagnostic applications.