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: 8097317
• 负责人: RICHARD L STOUFFER
• 金额: $ 66.18万
• 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 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8097317
• 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; Regimen; Reproductive Process; Research; Research Personnel; Rodent; Role; Sampling; Science; Services; Site; Somatic Cell; Steroids; Survivors; System; Testis; Tissue Engineering; Training; Training Programs; Transplantation; 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.

虽然现代的、积极的治疗方法使用电离放射疗法和化学疗法可以摧毁癌症， 它们可能具有持久的副作用，包括消除睾丸和卵巢中的生殖细胞， 不孕与有临床证明的选择的年轻男性不同， 年轻妇女和女孩仍然处于实验状态。该项目的长期目标是验证以下选项 通过防止卵巢暴露于以下物质的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）和返回配子或胚胎生育后， 根除癌症将在非人灵长类动物中进行转化研究，以：（目的1） 生物工程支架，支持灵长类动物卵泡的三维（3-D）结构， （2）在体外培养条件下，卵泡壁和卵母细胞的协调发育; 促性腺激素和生长因子在三维培养中促进卵泡生长和卵母细胞质量; (Aim 3）优化卵巢皮质自体移植的条件，以协调卵泡生长， 卵母细胞体内成熟;（目的4）评价卵母细胞体外受精和胚胎发育潜能 从体外成熟的卵泡和体内移植的卵巢皮质卵泡。不成熟的卵泡将 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 组件，如成功地用于啮齿动物卵泡和外推到灵长类动物，以允许生长。 将添加各种内分泌和局部因素，以确定卵泡生长的适当环境 （体细胞增殖，腔形成，卵泡直径）和分化（卵泡膜发育， 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容易接近的部位， 促进卵巢功能的恢复，即，将监测窦卵泡生长和月经周期。 最后，收集体外卵泡和体内皮质移植产生的成熟卵母细胞， 通过体外受精，胚胎移植到替代猕猴母亲体内， 怀孕开始和后代的健康。该项目将通过与Oncofertility的互动来促进 联合体，包括L博士。Shea和西北大学的生物材料核心P30 A，以及 向J. Critser博士和RO 1A提供灵长类动物样本冷冻保存平行研究的样本 密苏里州大学。促进体外灵长类卵泡生长的预期进展， 或者优化卵巢皮质移植和体内生长，将迅速转移到 由T博士指导的临床工作。伍德拉夫和生育中心网站在RO 1C和国家物理学家 P30 B合作，促进女性癌症幸存者的生育能力恢复。