Cryopreservation and Transplantation of Ovarian Cortical Tissue for Fertility Preservation

卵巢皮质组织的冷冻保存和移植以保存生育能力

• 批准号: 9288194
• 负责人: Mary B Zelinski
• 金额: $ 62.72万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-06 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9288194
• 关键词: 3-Dimensional; Adolescent; Adolescent and Young Adult; Adult; Adverse effects; Age; Antral; Autologous; Autologous Transplantation; Benign; Biological Preservation; Biomedical Engineering; Birth; Blood Vessels; Blood flow; Bone Marrow Transplantation; Breast Cancer Patient; Cancer Survivor; Child; Clinic; Clinical; Complex; Computer Simulation; Controlled Study; Cortex of ovary; Cryopreservation; Detection; Development; Diagnosis; Disease; Embryo; Embryology; Embryonic Development; Encapsulated; Endocrine; Environment; Exposure to; FDA approved; Female; Fertility; Fibrin; Foundations; Freezing; Future; Germ Cells; Goals; Health; Heterotopic Transplantation; Human; In Vitro; Individual; Infertility; Interruption; Investigation; Investigational Therapies; Late Effects; Macaca mulatta; Malignant Neoplasms; Menopause; Methods; Modeling; Mothers; Non-Malignant; Oocytes; Ovarian; Ovarian Tissue; Ovary; Parents; Patients; Population; Premature Menopause; Premature Ovarian Failure; Prior Therapy; Production; Protocols documentation; Radiation; Radiation therapy; Regulation; Reporting; Reproducibility; Research Personnel; Risk; Running; Safety; Site; Source; Stromal Cells; Survival Rate; Survivors; System; Techniques; Technology; Tissues; Translations; Transplantation; Transplanted tissue; Ultrasonography; Woman; cancer cell; cancer invasiveness; cancer therapy; cell type; chemotherapy; child bearing; childhood cancer survivor; clinical practice; clinical translation; contrast enhanced; cryobiology; experience; functional outcomes; girls; improved; in vivo; innovative technologies; nonhuman primate; novel; novel strategies; offspring; ovarian failure; ovary transplantation; pre-clinical research; preimplantation; prepuberty; public health relevance; restoration; scaffold; sound; steroid hormone; tool; treatment effect; vascular bed; young adult; young woman

 DESCRIPTION (provided by applicant): The long-range goals of this proposal are to develop robust and reproducible methods for ovarian tissue cryopreservation and transplantation as options for fertility preservation in survivors of cancer or other benign conditions wherein ovaria function is compromised. While 80% of young patients currently survive their cancers, devastating side effects of chemo- or radiotherapies leave female survivors facing infertility due to ovarian failure. Cryopreservation of ovarian tissue remains the only option for prepubertal, adolescent and young women with cancer as well as adult patients for whom immediate cancer therapy is required. But, human ovarian tissue cryopreservation is considered experimental and hampered by several factors. Embryology clinics in the U.S. favor embryo and oocyte vitrification over slow-rate freezing, but there is no uniform protocol for vitrification of ovaria tissue. Although 30 human births have been reported from slow-rate freezing and transplantation to orthotopic sites, only 4 could claim that the transplanted tissue was known to be the source of the oocytes. Most young survivors will not have a healthy ovarian vascular bed as a transplantation site following cancer therapies. Cryopreservation is also the only option for patients contraindicated for ovarian transplantation due to the risk of reintroducing malignant cells. While in vitro follicle culture is under development, most studies use follicles isolated frm non-cryopreserved tissues. We successfully devised a method for vitrification of rhesus monkey ovarian tissue in a closed system, and demonstrated post-thaw function in vivo and in vitro. When transplanted to heterotopic sites, vitrified tissue resumes ovarian function, and yields mature oocytes capable of preimplantation embryonic development. Secondary follicles isolated from vitrified tissue and encapsulated for 3-dimensional (3D) culture survive, grow to antral stages and produce steroid hormones in vitro, but oocyte quality is poor. Since the ovarian cortex is composed of complex cell types that complicate cryopreservation, vitrification of individual follicles and subsequent 3D culture is a novel approach toward preserving gametes for young patients with highly invasive cancers. An elite panel of co-investigators and collaborators that bring expertise in both state-of-the-art cryobiology as well as pre-eminent experience in clinical ovarian transplantation will help us advance the current technology for more rapid translation to clinical practice for patients. Using the rhesus monkey, we propose to identify the transplantation site for optimal fertile function of both prepubertal and young adult ovarian cortical tissue, improve long-term transplant function via local administration of factors important for revascularization as well as follicular survival, and optimize vitrification of isolaed preantral follicles and their ability to yield competent oocytes following 3D culture in vitro. Liv births in nonhuman primates after these controlled studies will establish the safety and feasibilit of experimental therapies prior to clinical translation so girls and young women can be offered the best chances of becoming mothers after surviving cancer.

 描述（由申请人提供）：该提案的长期目标是开发用于卵巢组织冷冻保存和移植的稳健且可重复的方法，作为癌症或卵巢功能受损的其他良性病症幸存者的生育力保存的选择。虽然目前 80% 的年轻患者在癌症中幸存下来，但化疗或放疗的毁灭性副作用使女性幸存者面临因卵巢功能衰竭而导致的不孕。对于患有癌症的青春期前、青少年和年轻女性以及需要立即进行癌症治疗的成年患者来说，卵巢组织的冷冻保存仍然是唯一的选择。但是，人类卵巢组织冷冻保存被认为是实验性的，并受到多种因素的阻碍。美国的胚胎学诊所更倾向于胚胎和卵母细胞玻璃化冷冻而不是慢速冷冻，但卵巢组织玻璃化冷冻没有统一的方案。尽管已经报道了 30 例通过慢速冷冻和移植到原位部位的人类分娩，但只有 4 例可以声称已知移植的组织是卵母细胞的来源。大多数年轻幸存者在癌症治疗后不会有健康的卵巢血管床作为移植部位。由于存在再次引入恶性细胞的风险，冷冻保存也是禁忌卵巢移植的患者的唯一选择。虽然体外卵泡培养正在开发中，但大多数研究使用从非冷冻保存组织中分离的卵泡。我们成功设计了一种在封闭系统中玻璃化恒河猴卵巢组织的方法，并在体内和体外证明了解冻后的功能。当移植到异位部位时，玻璃化组织恢复卵巢功能，并产生能够进行植入前胚胎发育的成熟卵母细胞。从玻璃化组织中分离并封装用于 3 维 (3D) 培养的次级卵泡能够存活、生长到窦期并在体外产生类固醇激素，但卵母细胞质量较差。由于卵巢皮质由复杂的细胞类型组成，使冷冻保存变得复杂，因此单个卵泡的玻璃化冷冻和随后的 3D 培养是一种为患有高侵袭性癌症的年轻患者保存配子的新方法。由共同研究人员和合作者组成的精英小组带来了最先进的冷冻生物学专业知识以及临床卵巢移植方面的卓越经验，将帮助我们推进当前的技术，以便更快地转化为患者的临床实践。利用恒河猴，我们建议确定青春期前和年轻成人卵巢皮质组织最佳生育功能的移植位点，通过局部施用对血运重建和卵泡存活重要的因子来改善长期移植功能，并优化分离的腔前卵泡的玻璃化冷冻及其在体外3D培养后产生有能力卵母细胞的能力。在这些对照研究之后，非人类灵长类动物的丽芙分娩将在临床转化之前确定实验疗法的安全性和可行性，以便女孩和年轻女性在癌症幸存后成为母亲的最佳机会。