Trial Information

Benign Reproductive Tissue Evaluation (BRTE) Study

Our hypothesis is that silent molecular lesions , defined as molecular alterations
detectable in histologically normal endometrial and ovarian tissues, represent markers of
cancer risk. Loss of expression of the PTEN tumor suppressor gene is currently the leading
marker of interest, based on the identification of clonal mutations in approximately 50% of
endometrial cancers and their precursors and the demonstration of small foci of PTEN-null
glands in approximately 40% of histologically unremarkable endometrial biopsies. This
proposal outlines a pilot study that will generate data necessary to design and conduct a
full-scale study to test the specific hypothesis that loss of PTEN expression in normal
endometrium accounts for a substantial percentage of risk conferred by known endometrial
cancer risk factors. Incessant ovulation represents one of the most widely-recognized
models to explain the pathogenesis of ovarian cancer, with women who have had a high number
of lifetime ovulatory menstrual cycles being at increased cancer risk because of repeated
ovulation-related injury to, and repair of, ovarian surface epithelium (OSE). This
extremely delicate single layer of cells exfoliates easily on handling, with the majority of
cells typically being lost in routine handling, when collected post-operatively.
Furthermore, the identification of early stage ovarian cancer is uncommon, and the vast
majority of ovarian cancers are not associated with recognizable precursors. The lack of
effective techniques for collecting and studying OSE in the laboratory represents a major
barrier to molecular studies designed to uncover the etiology and early pathogenesis of
ovarian cancer. This proposal will develop a collection method for OSE, and demonstrate its
utility for various molecular analyses. If successful, the collection of OSE will provide
the basis for larger studies aimed at identifying early molecular events in ovarian
carcinogenesis.

In this pilot, we will conduct endometrial and ovarian tissues (that would otherwise have
been discarded without histopathologic examination) from 125 hysterectomy and/or unilateral
or bilateral oophorectomy specimens obtained from women ages 18 to 54 years who were
operated on for benign indications. As an amendment to this active protocol, we propose
demonstrating the feasibility of obtaining intra-operative cytobrushings of ovarian surface
epithelial cells on 50 women to be accrued onto the study, which will include women having
hysterectomy (or unilateral oophorectomy) alone without removal of the ovaries at the time
of surgery.

We will administer a questionnaire assessing endometrial and ovarian cancer risk factors and
gynecologic history; obtain blood and urine, for future use in hormone studies; and obtain
carefully-mapped frozen and fixed endometrial and ovarian tissues. We will immunostain
endometrial tissues to assess the presence, number, location, and size of foci containing
PTEN-null glands, which represents a validated surrogate of mutations in the PTEN tumor
suppressor gene. This pilot will demonstrate the feasibility of executing this complex
protocol; determine the number and spacing of sections required to accurately and
efficiently assess the PTEN status of the endometrium; and provide data for developing power
estimates needed to propose a full-scale study with a sufficient number of subjects to test
our hypothesis that PTEN abnormalities account for a substantial proportion of the risk
associated with recognized epidemiologic endometrial cancer risk factors. It will assess
the feasibility of performing molecular analyses on ovarian surface epithelial cells
collected intra-operatively, and correlating molecular finding with known ovarian cancer
risk factors. If successful, this will provide the basis for larger studies aimed at
identifying early molecular events in ovarian carcinogenesi, particularly in the setting of
women at increased genetic risk of ovarian cancer. The pilot itself will also provide an
extremely valuable repository for future biomarker pilot studies.