Whole Ovary Vitrification for Fertility Preservation

项目摘要 该提案的长期目标是开发一种安全、有效和用户友好的玻璃化系统， 冷冻保存整个人类卵巢。在美国，超过140 000名0-49岁的女孩和妇女被诊断为 每年的癌症。虽然诊断和治疗的改善提高了生存率（75%）， 绝经前和85%的儿童癌症患者），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。胚胎冷冻保存时 而卵母细胞是有效的，卵巢皮质和整个卵巢冷冻保存是未来生育的唯一选择 青春期前患者和需要立即进行癌症治疗的患者。冷冻保存整个卵巢， 与卵巢组织相比，具有以下优点：1）保留所有类型的卵泡， 对于体外成熟，2）更长的移植物寿命，和3）移植后由现有血管支持的更快恢复。 血管吻合移植然而，目前用于全卵巢冷冻保存的方法是 次优，并且没有设计用于冷冻保存人类卵巢的技术，尽管它是最好的 青春期前癌症患者未来生育能力和完整内分泌功能的选择， 外源激素的帮助。这项研究是世纪医学更广泛项目的一部分 开发上级器官冷冻保存系统，包括冷冻保护剂（CPA）溶液， 用于CPA装载和卸载的灌注装置（硬件和软件）和液氮储存单元。 第二阶段将帮助我们开始努力实现我们作为器官银行公司的最终目标。我们 公司已经成功地证明了玻璃化冷冻兔肾移植后的功能保存。 在这项SBIR II期研究中，我们将验证并继续改善我们迄今为止在全卵巢中的成功 玻璃化冷冻使用猪和绵羊中卵巢移植的大动物模型（绵羊具有已建立的 卵巢移植手术，它们的卵巢结构比猪更接近人类）。我们还将 开发用于卵巢移植的非人灵长类动物临床前模型，并表征低温 猴子卵巢的特性此外，我们将最终确定卵巢灌注装置，并开发一个封闭的 用于储存个体冷冻保存的卵巢以供最终临床使用的系统。由此产生的技术 二期项目将是第一个为人类卵巢设计的灌注装置，可以同时控制 温度、压力、流速和CPA浓度，并具有内置反馈机制， 在灌注期间保持恒定压力。在第二阶段成功完成后，我们将提交第二阶段b 申请纳入使用非人灵长类动物模型、人体组织异种移植的生育力试验 实验，并最终确定商业化战略。这一新的系统将为人类提供新的希望。 数以千计的癌症幸存者，其影响可能是重大和持续的。

PROJECT SUMMARY The long-term goal of this proposal is to develop a safe, effective and user-friendly vitrification system for cryopreservation of whole human ovaries. In the U.S., over 140,000 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. While cryopreservation of embryos and oocytes are effective, ovarian cortex and whole ovary cryopreservation is the only option for future fertility in prepubertal patients and those who require immediate cancer treatment. Cryopreservation of whole ovaries, compared to ovarian tissue, has added advantages of 1) preserving all classes of follicles, which can be used for in vitro maturation, 2) longer graft lifespan, and 3) faster recovery supported by existing blood vessels after transplantation with vascular anastomosis. However, current methods for whole ovary cryopreservation are suboptimal and there is no technology designed for cryopreserving human ovaries, despite it being the best choice for prepubertal cancer patients for future fertility and intact endocrine function for decades without the help of exogenous hormones. The proposed research is part of 21st Century Medicine’s more general program of developing superior organ cryopreservation systems, including cryoprotective agent (CPA) solutions, perfusion apparatus (hardware and software) for CPA loading and unloading, and liquid nitrogen storage units. This Phase II will help us initiate our effort in achieving our ultimate goal as an organ banking company. Our company has successfully demonstrated functional preservation of vitrified rabbit kidneys after transplantation. In this SBIR Phase II study, we will validate and continue improving our success thus far in whole ovary vitrification using large animal models of ovary transplantation in pigs and sheep (sheep has an established ovary transplant procedure and their ovarian structure is closer to the human than the pig). We will also develop a nonhuman primate preclinical model for ovary transplantation as well as characterize the cryogenic properties of monkey ovaries. Furthermore, we will finalize the Ovary Perfusion Device and develop a closed system for storing individual cryopreserved ovaries for eventual clinical use. The resultant technology of this Phase II project will be the first perfusion unit designed for the human ovary and can simultaneously control temperature, pressure, flow rate and CPA concentration with a build-in feedback mechanism to maintain constant pressure during perfusion. Upon successful completion of Phase II, we will submit a Phase IIb application to include fertility trials using the nonhuman primate model, human tissue xenografting experiments, and finalizing strategies for commercialization. This novel system will offer new hope to the thousands of cancer survivors and the impact could be significant and sustained.