Optimized protocols for ovarian tissue cryopreservation and novel modRNA-based approaches to improve transplantation outcomes for fertility preservation in chronically diseased girls

项目摘要 随着诊断精度的提高和治疗选择的扩大，儿童的健康和寿命 出生时患有慢性病的人已经显著延长到成年，这引起了人们对保护他们的 生活质量和生殖选择。不幸的是，许多慢性病的病理可能有 对性腺产生负面影响，导致不孕和/或过早绝经。 整个卵巢组织的冷冻保存(以及后来的自体移植)可以提供一种“安全的”方法. 保护卵巢储备免受疾病病理的性腺毒性影响，这种方法一直以来 自十多年前的早期尝试以来，该技术在癌症患者中的应用至少产生了100例活产。 尽管这种方法是有效的，但冷冻保存的卵巢组织的活力和生产力是 急性期移植物内的炎性、缺血环境严重破坏 移植后。 该方案利用了强大的生物活性和直接传递修饰的mrna。 (ModRNA)作为一种提高自体卵巢组织移植物活力和产量的治疗方法。 使用我们团队开发的异种移植平台，我们将研究modRNA的潜力：1) 提高卵巢组织移植的存活率和产量，以保存生育能力；2) 在静止的原始状态下维持一个强大的卵泡池，这将作为一个长期的 内分泌功能和卵母细胞输出正常化的储存库；以及3)证明了这一方法的有效性 在高度相关的临床前动物模型恒河猴中的方法。通过这些实验，我们将 为稳健、可重复和FDA批准的卵巢组织治疗方法奠定基础 冷冻保存/自体移植在希望保持生育能力的慢性疾病女孩中的应用 选择。

Project Summary With increased diagnostic precision and expanded treatment options, the health and lifespan of children born with chronic diseases has extended significantly into adulthood, drawing attention to preserving their quality of life and reproductive options. Unfortunately, the pathology of many chronic conditions can have negative consequences for the gonad, resulting in infertility and/or premature menopause. Cryopreservation (and later auto-transplantation) of whole ovarian tissue could provide a means of “safe- guarding” ovarian reserve from the gonadotoxic effects of disease pathology, and this approach has been utilized in cancer patients to yield at least 100 live births since early attempts more than a decade ago. Although this approach is effective, the viability and productivity of cryopreserved ovarian tissue is drastically undermined by the inflammatory, ischemic environment within the graft during the acute phase post-transplant. This proposal leverages the potent bioactivity and straightforward delivery of modified mRNA (modRNA) as a therapeutic approach to improving the viability and output of ovarian tissue autografts. Using a xenograft platform developed by our group, we will investigate the potential for modRNA to: 1) improve viability and output of ovarian tissue transplanted for the purpose of fertility preservation; 2) maintain a robust pool of ovarian follicles in a quiescent primordial state that will serve as a long-term reservoir for normalization of endocrine function and oocyte output; and 3) demonstrate the efficacy of this approach in a highly relevant pre-clinical animal model, rhesus macaque. With these experiments we will establish a foundation for robust, repeatable and FDA-approvable methods for ovarian tissue cryopreservation/auto-transplantation in chronically diseased girls that wish to maintain their reproductive options.