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

用于在药物开发过程中筛选性腺毒性的离体全卵巢培养系统

• 批准号: 10823136
• 负责人: Alison Y Ting
• 金额: $ 30.93万
• 依托单位: EXPANSE BIO LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10823136
• 关键词: 3-Dimensional; Age; Animals; Apoptosis; Back; Basic Science; Biological Assay; Blood Vessels; Bone Diseases; Cancer Patient; Carbon Dioxide; Cardiovascular Diseases; Cattle; Cells; Cessation of life; Clinical Research; Cognition Disorders; Collaborations; Collection; Computer software; Culture Media; Cyclophosphamide; Cytotoxic agent; Development; Devices; Diagnosis; Drug Screening; Drug usage; Endocrine; Ethics; Evaluation; Exposure to; Family suidae; Feedback; Female; Fertility; Future; Gene Expression Profiling; Genetic Transcription; Growing Follicle; Health; Histology; Hour; Human; Immunohistochemistry; In Vitro; Incubators; Industrialization; Infertility; Lactic acid; Laminin; Malignant Childhood Neoplasm; Malignant Neoplasms; Mammals; Measurement; Menopause; Metabolic; Microfluidics; Modeling; Molecular; Morphology; Nutrient; Oncologist; Oocytes; Organ; Ovarian; Ovarian Follicle; Ovary; PECAM1 gene; Patients; Perfusion; Pharmaceutical Preparations; Phase; Premature Menopause; Premature Ovarian Failure; Premenopause; Protocols documentation; Quality of life; Risk; Rodent; Rodent Model; Sheep; Small Business Innovation Research Grant; Structure; Survival Rate; System; Technology; Testing; Therapeutic; Therapeutic Agents; Tissue Banks; Tissues; Toxic effect; Transplantation; Woman; autocrine; blood damage; cancer diagnosis; cancer therapy; cell type; chemotherapeutic agent; chemotherapy; commercialization; cost effective; cytotoxic; design; dosage; drug candidate; drug development; experience; folliculogenesis; girls; human model; improved; in vivo; meter; novel; novel therapeutics; ovarian damage; ovotoxicity; paracrine; phase 2 study; porcine model; pressure; protein expression; screening; side effect; steroid hormone; three dimensional cell culture; timeline; tissue/cell culture; treatment planning; wasting; young cancer survivor; young woman

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.

摘要 背景：在美国，每年有20多万0-49岁的女孩和妇女被诊断出患有癌症。而 诊断和治疗的改善提高了生存率（绝经前75%，绝经后85%）。 儿童癌症患者），癌症治疗的毁灭性副作用包括卵巢早衰， 不孕症和更年期相关的健康风险。检测和消除风险的现行行业标准 卵毒性是使用体内啮齿动物模型或2D/3D体外卵巢细胞/组织培养具有显著缺点。 该I期提案旨在开发一种体外系统，用于长期评估卵巢损伤， 形态学、功能和分子水平，在整个卵巢模型中。 方法：该第一阶段SBIR提案的目标是：1）构建一种新型的离体微流体器官 灌注装置，其将支持整个卵巢培养7天并建立最佳培养条件， 以及2）使用化疗药物4-氢过氧环磷酰胺， 最常用的药物之一环磷酰胺的代谢产物。终点将包括以下能力： 卵母细胞在体外成熟，以及证明卵泡健康的标志物的蛋白质和RNA表达 和凋亡。该项目的最终技术将是第一个为整个系统设计的灌注装置。 大型哺乳动物（包括人类）的卵巢，并可以控制灌注压力，流速和治疗 通过内置的反馈机制来控制浓度，以在灌注期间保持恒定的压力。 未来方向和商业化潜力：第一阶段项目将提供关键的原理证明 对于未来的II期研究，以测试整个卵巢培养系统培养更长时间（1个月）的能力， 与制药公司合作，筛选具有卵毒性或卵保护潜力的候选药物， 通过移植化疗处理的卵巢来检查长期内分泌和生育能力的后果 回到宿主动物身上维持延长的生命/功能的技术具有额外的潜力， 观察卵泡成熟及其相关研究（基础和临床研究）。成功开发一个 离体全卵巢培养系统将在药物开发期间设定新的工业标准，因为它 允许使用大型哺乳动物（包括女性）的卵巢，而不会对动物造成伤害和死亡， 并且它将允许评价整个卵巢，同时模拟毒性化疗剂的体内递送。