Objectives.

The objective of this project is to further develop our organotypic model of epithelial ovarian tissue and endometrial tissue into models of carcinogenesis that can be used to study the mechanisms of carcinogenesis and chemoprevention.

Organotypic Cultures.

Monolayer cultures of cells are not accurate representations of in vivo tissue because they lack the types of interactions that cells have with other cells and with the extracellular matrix in vivo. These interactions are known to influence gene expression and thus the constitution and behavior of cells. Organotypic cultures are tissue culture models that mimic in vivo tissue architecture through manipulation of epithelial and stromal cells within and on top of an extracellular matrix. A benefit of organotypic cultures is that they incorporate aspects of cell-matrix and epithelial-stromal interactions that cannot be evaluated in monolayer cultures. Another benefit is that they can be used to evaluate monoclonal colonies, whereas individual xenograft tumors in animals are established from millions of cells. Colonies in organotypic culture therefore, more accurately mimic tumorigenesis than animal xenografts. Also, organotypic cultures can be more readily manipulated than animal models allowing increased numbers of experiments and less cost. Organotypic cultures however cannot be used to replace animal models because the cultures do not incorporate metabolic, physiologic and immunologic effects. Thus organotypic cultures have the potential to be used as a screening tool to identify the best chemoprevention agents for further testing in animal models. In addition, they provide opportunities to evaluate hypothesis driven research on carcinogenesis and chemoprevention. Current organotypic models however, only provide static representations of normal, premalignant or carcinogenic tissue, and do not incorporate the process of carcinogenesis.

Current Progress.

We have been able to consistently transform normal endometrial organotypic cultures into the cancerous phenotype and prevent this change with our lead compound SHetA2. We are in the process of identifying the molecular changes that occur.