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: 7502587
• 负责人: RICHARD L STOUFFER
• 金额: $ 65.14万
• 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/7502587
• 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; Condition; Cortex of ovary; Cryopreservation; Development; Education; Embryo; Embryo Transfer; Endocrine; Estrogens; Excision; Exposure to; Extracellular Matrix; Female; Fertility; Fertilization; Fertilization in Vitro; Follicle Stimulating Hormone; Forearm; Germ Cells; Goals; Gonadal structure; Graafian Follicles; Growth; Growth Factor; Health; Health Services Research; Hormones; Hydrogels; In Vitro; Infertility; Live Birth; Macaca; Macaca mulatta; Malignant Neoplasms; Meiosis; Methods; Missouri; Modeling; Monitor; Monkeys; Mothers; Mus; Numbers; Oocytes; Ovarian; Ovary; Patients; Periodicity; Pregnancy; Primates; Quality of life; Radiation therapy; Reproductive Process; Research; Research Personnel; Rodent; Role; Sampling; Science; Site; Somatic Cell; Steroids; Survivors; System; Testis; Tissue Engineering; Training; Training Programs; Transplantation; Treatment Protocols; Universities; Vascular Endothelial Growth Factors; Woman; cancer type; chemotherapy; concept; design; egg; girls; in vivo; interdisciplinary approach; member; men; neovascularization; nonhuman primate; novel; oocyte maturation; 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.

尽管使用电离放射疗法和化学疗法的现代积极治疗可以消灭癌症， 它们可能会产生持久的副作用，包括消除睾丸和卵巢中的生殖细胞，因此 不孕症。与拥有经过临床证明的选择的年轻男性不同，保留生育能力的方法 年轻妇女和女孩仍处于实验阶段。该项目的长期目标是验证以下选项 通过防止卵巢暴露于配子毒性作用来恢复女性癌症幸存者的生育能力 治疗（通过去除和保存活检）并返回配子或胚胎以实现生育能力 根除癌症。将在非人类灵长类动物中进行转化研究，以：（目标 1） 生物工程支持灵长类动物卵泡三维 (3-D) 结构的支架 允许卵泡壁和卵母细胞在体外协调发育； （目标 2）评估角色 促性腺激素和生长因子在 3D 培养过程中促进卵泡生长和卵母细胞质量； （目标 3）优化卵巢皮质自体移植条件，以协调卵泡生长和 卵母细胞体内成熟； （目标 4）评估卵母细胞的受精和胚胎潜力 来自体外成熟卵泡和体内移植的卵巢皮质卵泡。未成熟的卵泡将会 从恒河猴卵巢中分离并添加到藻酸盐水凝胶和内源性 ECM 的 3-D 基质中 成分，已成功用于啮齿动物毛囊并外推到灵长类动物以允许生长。 将添加各种内分泌和局部因素来确定卵泡生长的适当环境 （体细胞增殖、窦形成、卵泡直径）和分化（鞘膜发育、 雌激素分泌），以及其封闭的卵丘-卵母细胞复合体的成熟。卵巢皮质 样本将被自体移植到易于接近的部位，并且促血管生成因子的能力 促进卵巢功能的恢复，即窦卵泡生长和月经周期，将受到监测。 最后，收集体外卵泡和体内皮质移植产生的成熟卵母细胞并 通过体外受精、胚胎移植到代孕猕猴母亲体内评估生殖潜力， 怀孕开始和后代的健康。与 Oncofertility 的互动将促进该项目 联盟成员包括 L. Shea 博士和西北大学生物材料核心 P30A，以及 向 J. Critser 博士和 RO1A 提供用于灵长类动物样本冷冻保存平行研究的样本 密苏里大学。促进灵长类卵泡体外生长的预期进展，以及 或者优化卵巢皮质移植和体内生长，将迅速转移到 由 T. Woodruff 博士和 RO1C 生育中心以及国家医师指导的临床工作 与 P30B 合作，促进女性癌症幸存者的生育能力恢复。