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.
Public Health Relevance Statement: PROJECT NARRATIVE One devastating side effect of chemotherapeutics is destruction of ovarian follicles which can result in infertility and early menopause in girls and young women. An ex vivo system for long-term whole ovary culture that can help screening and de-risking chemotherapeutics for their potential ovotoxicity does not currently exist. The objectives for this Phase I SBIR proposal are to build a novel ex vivo whole ovary culture system, establish optimal perfusion and culture conditions, as well as to test this system using a chemotherapy medication, 4- hydroperoxy cyclophosphamide, with known ovotoxic effects.
Project Terms: ages; Age; Animals; Back; Dorsum; Biological Assay; Assay; Bioassay; Biologic Assays; Blood Vessels; vascular; Bone Diseases; bone disorder; Malignant Neoplasms; Cancers; Malignant Tumor; malignancy; neoplasm/cancer; Carbon Dioxide; CO2; Carbonic Anhydride; Cardiovascular Diseases; cardiovascular disorder; Cattle; Bovine Species; bovid; bovine; cow; Cells; Cell Body; Clinical Research; Clinical Study; Cognition Disorders; cognitive disease; cognitive disorder; cognitive syndrome; Culture Media; growth media; Cyclophosphamide; CTX; CYCLO-cell; Carloxan; Ciclofosfamida; Ciclofosfamide; Cicloxal; Clafen; Claphene; Cycloblastin; Cycloblastine; Cyclophospham; Cyclophosphamidum; Cyclophosphan; Cyclophosphane; Cyclophosphanum; Cyclostin; Cyclostine; Cytophosphan; Cytophosphane; Cytoxan; Endoxan; Endoxana; Enduxan; Fosfaseron; Genoxal; Genuxal; Ledoxina; Mitoxan; Neosar; Procytox; Sendoxan; Syklofosfamid; Zytoxan; Cessation of life; Death; Diagnosis; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Ethics; ethical; Feedback; Female; Fertility; Fecundability; Fecundity; Future; Ovarian Follicle; ovarian follicular pool; Health; Histology; Human; Modern Man; Immunohistochemistry; Immunohistochemistry Cell/Tissue; Immunohistochemistry Staining Method; In Vitro; Incubators; Industrialization; Infertility; Cannot achieve a pregnancy; Difficulty conceiving; fertility cessation; fertility loss; infertile; Laminin; Glycoprotein GP-2; Mammals; Mammalia; Menopause; Premature Menopause; early menopause; Oocytes; Ovocytes; Ovary; Patients; Perfusion; pressure; Quality of life; QOL; Risk; Rodent; Rodentia; Rodents Mammals; Sheep; Ovine; Ovis; Computer software; Software; Survival Rate; Family suidae; Pigs; Suidae; Swine; porcine; suid; Technology; Testing; Tissue Banks; Tissue Collection; Tissue repository; Tissues; Body Tissues; Genetic Transcription; Gene Transcription; RNA Expression; Transcription; Transplantation; transplant; Woman; Lactic acid; premature ovarian aging; Premature Ovarian Failure; timeline; Apoptosis Pathway; Programmed Cell Death; Apoptosis; dosage; Organ; improved; Ovarian; Phase; Pre-Menopause; Pre-menopausal Period; Premenopausal; Premenopausal Period; pre-menopausal; premenopausal status; Premenopause; Evaluation; wasting; Measurement; drug use; Drug usage; Oncologist; Childhood Cancers; Malignant Childhood Tumor; Malignant Pediatric Neoplasm; Malignant Pediatric Tumor; Malignant childhood cancer; cancer in a child; cancer in children; child with cancer; childhood malignancy; children with cancer; pediatric cancer; pediatric malignancy; Malignant Childhood Neoplasm; Collaborations; Therapeutic Steroid Hormone; steroid hormone; Therapeutic; Therapeutic Agents; Cytotoxic drug; Cytotoxic agent; Metabolic; Exposure to; Morphology; Hour; Protocols documentation; Protocol; cell type; System; 3-Dimensional; 3-D; 3D; three dimensional; meter; Endocrine; autocrine; Autocrine Systems; experience; paracrine; treatment planning; Toxic effect; Toxicities; Structure; Nutrient; novel; Basic Science; Basic Research; Devices; chemotherapeutic agent; Gene Expression Profiling; Gene Expression Monitoring; Gene Expression Pattern Analysis; Transcript Expression Analyses; Transcript Expression Analysis; analyze gene expression; gene expression analysis; gene expression assay; transcriptional profiling; Modeling; girls; drug development; cancer therapy; Cancer Treatment; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; anti-cancer therapy; anticancer therapy; cancer-directed therapy; cancer diagnosis; protein expression; µfluidic; Microfluidics; tissue/cell culture; PECAM1 gene; CD31; PECAM1; cytotoxic; Growing Follicle; in vivo; Cancer Patient; Collection; Rodent Model; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Molecular; Development; developmental; designing; design; ovarian toxicity; ovotoxicity; cost effective; folliculogenesis; chemotherapy; new drug treatments; new drugs; new pharmacological therapeutic; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel pharmaco-therapeutic; novel pharmacological therapeutic; novel therapy; novel therapeutics; commercialization; drug candidate; phase II study; phase 2 study; adolescent woman; adolescent women; young woman; screenings; screening; young cancer survivor; 3D cell culture; 3D culture; three dimensional cell culture; Drug Screening; model of human; human model; ovarian damage; blood damage; side effect; pig model; piglet model; swine model; porcine model
