Ex vivo whole ovary culture system for screening gonadotoxicity during drug development

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ABSTRACT Background: In the U.S., over 200,000 girls and women age 0-49 are diagnosed with cancer annually. While improved diagnosis and treatment have led to increased survival rates (75% in premenopausal and 85% in childhood cancer patients), devastating side effects of cancer therapies include premature ovarian failure, infertility and menopause-related health risks. The current industrial standard for detecting and de-risking ovotoxicity is to use in vivo rodent models or 2D/3D in vitro ovarian cells/tissue culture with significant drawbacks. This Phase I proposal aims to develop an ex vivo system for long-term assessment of ovarian damage at morphological, functional, and molecular levels, in a whole ovary model. Approach: The objectives for this Phase I SBIR proposal are to 1) build a novel ex vivo microfluidic organ perfusion device that will support culture of whole ovaries for 7 days and establish optimal culture conditions, as well as to 2) test this culture system using a chemotherapy medication, 4-hydroperoxy cyclophosphamide, a metabolite from one of the most commonly used drugs, cyclophosphamide. Endpoints will include the ability of oocytes to mature in vitro, as well as protein and RNA expression of markers that demonstrate follicular health and apoptosis. The resultant technology of this project will be the first perfusion unit designed for the whole ovary of large mammals (including humans) and can control perfusion pressure, flow rate and treatment concentration with a build-in feedback mechanism to maintain constant pressure during perfusion. Future Directions and Commercialization potential: This phase I project will provide crucial proof-of-principle for future Phase II studies to test the ability of the whole ovary culture system for longer culture period (1 month), collaborate with drug companies to screen drug candidates for their ovotoxicity or ovoprotection potentials, and to examine long-term endocrine and fertility consequences by transplanting chemotherapeutics-treated ovaries back to the host animal. The technology to maintain prolonged live/functional has the added potential for observing follicle maturation and its related studies (basic and clinical research). Successful development of an ex vivo whole ovary culture system will set new industrial standards during drug development because it will allow the use of ovaries from larger mammals (including women) without causing harm and death to the animal, and it will allow evaluation of the whole ovary while mimicking in vivo delivery of toxic chemotherapeutics.