Evaluation of Multiple Biomarkers to Estimate Risk of Ovarian Cancer in Patients With a Pelvic Mass.

This study is exploratory in nature and is designed to be hypothesis generating to support the design of future studies. A total of 200 women diagnosed with a pelvic mass (defined as a simple, complex or a solid ovarian cyst / pelvic mass) who are scheduled for a laparotomy or laparoscopy for removal of the pelvic mass will be enrolled for evaluation of the primary and exploratory endpoints. An initial evaluation of the data will be conducted after identification of 15 evaluable subjects with histopathologically confirmed ovarian cancer. It is estimated that 50 - 100 subjects will be required to obtain a minimum of 15 evaluable subjects with histopathologically confirmed ovarian cancer. The remainder of the women enrolled (expected to be ~100 - 150 subjects) will be used to verify the findings from the initial evaluation (i.e. correlation of the markers with the absence or presence of malignancy) and to refine the assay/algorithm.

Within 60 days prior to surgery, each subject must have a pelvic imaging study (e.g. ultrasound, CT scan, MRI, etc.) conducted and read to visualize the pelvic mass according to the current standard of care. Results of the pelvic imaging study(ies) will be recorded.

Within 60 days prior to, or on the day of the pelvic mass surgery, collect up to 35mL of peripheral blood into one 5mL SST tube, which must be drawn first, followed by three separate 10mL EDTA tubes. Serum from SST tube will be prepared at the local study center and used for protein biomarker testing. The EDTA tubes will be shipped to the Medical University of Vienna, where the blood will be pooled and equal volumes processed using two different separation methods on the Parsortix™ system to capture and harvest rare cells. The harvested material from the EDTA tubes will be lysed, and total RNA will be extracted from the cell lysate for evaluation of multiple gene targets using quantitative PCR (qPCR).

Laparotomy or laparoscopy for removal of the pelvic mass will be performed by a qualified individual. Tissue samples will be sent to the local pathology department for histological examination in accordance with standard institutional practices. Results of the histopathological evaluation will be recorded, including the final diagnosis along with histological sub-type, and if available, stage and grade of ovarian cancer where disease is identified.

Subjects will be considered negative for ovarian cancer:

• if the subject undergoes surgery and no mass is identified, or;
• if the histopathological findings are negative for cancer (i.e. benign conditions).

Subjects will be considered positive for ovarian cancer:

• if the histological examination of the tissue taken at the time of surgery confirms the presence of ovarian, primary peritoneal and/or fallopian tube cancers.

For the purposes of enrollment, subjects diagnosed with a malignancy other than an ovarian cancer as well as those diagnosed with low malignant potential (LMP) / borderline tumors will not be counted as an ovarian cancer. However, two separate analyses of the final study data will be conducted: one where subjects diagnosed with other cancers are excluded from the analysis of the primary endpoint and the subjects diagnosed with LMP / borderline tumors are classified as being negative for malignancy, and a second where subjects diagnosed with other cancers and those diagnosed with LMP / borderline tumors are classified as being positive for malignancy.

Demographical and clinical data may be summarized using descriptive statistics. Continuous variables may be summarized using the number of observations, mean, standard deviation, coefficient of variation, median, and range as appropriate. Categorical values may be summarized using the number of observations and percentages as appropriate.

The association of the markers (i.e. serum protein markers and mRNA markers) with the histopathological diagnosis will be assessed using appropriate statistical methods (e.g., logistic regression, analysis of variance [ANOVA], etc.), depending on the endpoints. Analyses may be performed within and between various histopathological diagnosis sub-groups. Other clinical covariates (such as imaging results and subject demographics) may also be included in the modeling.

An initial evaluation of the correlation of the markers with the histopathological diagnosis will be performed after the identification of 15 subjects with histopathologically verified ovarian cancer. The final cohort of patients will be used to verify the findings from the initial evaluation (i.e. correlation of the markers with the absence or presence of malignancy) and to refine the assay/algorithm. Bootstrap analyses may be utilized on the entire set of 200 subjects to finalize an assay/algorithm for further study.