RO1B: Bioengineering Primate Follicles: From Immature Eggs to Live Births (3/10)

RO1B：灵长类卵泡生物工程：从未成熟卵到活产 (3/10)

• 批准号: 7649502
• 负责人: RICHARD L STOUFFER
• 金额: $ 67.57万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649502
• 关键词: 3-Dimensional; Abdomen; Academy; Activins; Address; Adverse effects; Alginates; Angiogenic Factor; Animal Experimentation; Antral; Architecture; Assisted Reproductive Technology; Autologous Transplantation; Biocompatible Materials; Biological Preservation; Biology; Biomedical Engineering; Biopsy; Caliber; Cancer Survivor; Cell Proliferation; Cells; Clinical; Collagen; Complex; Cortex of ovary; Cryopreservation; Development; Education; Embryo; Embryo Transfer; Endocrine; Estrogens; Ethics; Excision; Exposure to; Extracellular Matrix; Female; Fertility; Fertilization; Fertilization in Vitro; Forearm; Germ Cells; Goals; Gonadal structure; Graafian Follicles; Growth; Growth Factor; Health; Hormones; Hydrogels; In Vitro; Infertility; Live Birth; Macaca; Macaca mulatta; Malignant Neoplasms; Meiosis; Methods; Missouri; Modeling; Monitor; Monkeys; Mothers; Mus; Oocytes; Ovarian; Ovary; Patients; Periodicity; Pregnancy; Primates; Quality of life; Radiation therapy; Reproductive Process; Research; Research Personnel; Rodent; Role; Sampling; Science; Services; Site; Somatic Cell; Steroids; Survivors; System; Testis; Tissue Engineering; Training; Training Programs; Transplantation; Treatment Protocols; Universities; Vascular Endothelial Growth Factors; Woman; cancer type; chemotherapy; design; egg; girls; in vivo; interdisciplinary approach; member; men; neovascularization; nonhuman primate; novel; offspring; oocyte maturation; preimplantation; prevent; reproductive; restoration; scaffold; subcutaneous; success; translational study; young adult

Although modern, aggressive treatments using ionizing radiotherapy and chemotherapy destroy cancers, they can have lasting side effects, including elimination of germline cells in the testes and ovary, and hence infertility. Unlike young men who have a clinically proven option, the methods for fertility preservation for young women and girls remain experimental. The long-term goal of this project is to validate options for restoring fertility in female cancer survivors by preventing ovarian exposure to the gametotoxic effects of therapy (by removing and preserving biopsies) and returning gametes or embryos for fertility after eradicating the cancer. Translational studies will be performed in nonhuman primates to: (Aim 1) Bioengineer a scaffold that supports the three-dimensional (3-D) architecture of the primate follicle and permits coordinated development of the follicle wall and oocyte in vitro; (Aim 2) Evaluate the role of gonadotropic hormones and growth factors in promoting follicle growth and oocyte quality during 3-D culture; (Aim 3) Optimize conditions for autotransplantation of ovarian cortex for coordinated follicle growth and oocyte maturation in vivo; and (Aim 4) Assess the fertilization and embryonic potential of oocytes derived from in vitro matured follicles and transplanted ovarian cortical follicles in vivo. Immature follicles will be isolated from rhesus macaque ovaries and added to a 3-D matrix of alginate hydrogel and endogenous ECM components, as successfully employed for rodent follicles and extrapolated to the primate to permit growth. Various endocrine and local factors will be added to determine the appropriate milieu for follicle growth (somatic celh proliferation, antrum formation, follicular diameter), and differentiation (thecal development, estrogen secretion), as well as maturation of its enclosed cumulus-oocyte complex. Ovarian cortical samples will be autotransplanted to readily accessible sites and the ability of pro-angiogenic factors to promote restoration of ovarian function, i.e., antral follicle growth and menstrual cyclicity, will be monitored. Finally, mature oocytes produced by follicles in vitro and cortical transplants in vivo will be collected and evaluated for reproductive potential by in vitro fertilization, embryo transfer into surrogate macaque mothers, pregnancy initiation and health of offspring. This project will be facilitated by interactions with Oncofertility Consortium and include Dr. L. Shea and the Biomaterials Core P30A at Northwestern University, and provide samples for parallel studies on cryopreservation of primate samples to Dr. J. Critser and RO1A at the University of Missouri. The expected advances in promoting primate follicle growth in vitro, and alternatively optimizing ovarian cortical transplantation and growth in vivo, will be transferred rapidly to clinical efforts directed by Dr. T. Woodruff and the Fertility Center sites in RO1C and the National Physcians Cooperative in P30B, to promote fertility restoration in female cancer survivors.

尽管使用电离放疗和化学疗法破坏癌症现代，积极的治疗，但 它们可以具有持久的副作用，包括消除睾丸和卵巢中的种系细胞，因此 不育。与具有临床证明选择的年轻人不同，保存生育的方法 年轻妇女和女孩继续实验。该项目的长期目标是验证选项 通过防止卵巢暴露于卵巢毒性作用来恢复女性癌症幸存者的生育能力 治疗（通过删除和保存活检）以及在 消除癌症。转化研究将在非人类灵长类动物中进行：（目标1） 生物工程师的脚手架，支持灵长类卵泡的三维（3-D）结构 允许体外的卵泡壁和卵母细胞的协调发展； （目标2）评估 3-D培养期间促进卵泡生长和卵母细胞质量的促性激素和生长因子； （目标3）优化卵巢皮层自动移植的条件，以协调卵泡生长和 体内卵母细胞成熟； （目标4）评估卵母细胞的施肥和胚胎潜力 从体外的体外成熟卵泡和体内移植的卵巢皮质卵泡。未成熟的卵泡将是 从猕猴卵巢中分离出来，并添加到藻酸盐水凝胶和内源性ECM的3-D基质中 成分，成功用于啮齿动物卵泡，并推送到灵长类动物以允许生长。 将添加各种内分泌和局部因素，以确定适当的卵泡生长环境 （体细胞增殖，胸骨形成，卵泡直径）和分化（thecal发育， 雌激素分泌），以及其封闭的积云元素复合物的成熟。卵巢皮质 样品将自动译为易于访问的地点以及亲血管生成因素的能力 将监测卵巢功能的恢复，即，将监测肛门卵泡生长和月经循环。 最后，将收集由卵泡在体外和皮质移植中产生的成熟卵母细胞，并收集体内的皮质移植物，并 评估通过体外受精，胚胎转移到替代猕猴中的生殖潜力， 后代的怀孕开始和健康。该项目将通过与OncoFertity的互动来促进 财团，包括西北大学的L. Shea博士和生物材料核心P30A，以及 为J. Critser博士和RO1A的冷冻保存提供了样品，以进行平行研究 密苏里大学。预期在体外促进灵长类动物卵泡生长的预期进展，以及 或者，在体内优化卵巢皮质移植和生长，将迅速转移到 由T. Woodruff博士和RO1C的生育中心站点和国家医生指导的临床工作 P30b的合作社，以促进女性癌症幸存者的生育能力。