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期，这引起了人们对保护他们的健康的关注。 生活质量和生殖选择。不幸的是，许多慢性疾病的病理学可能具有 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（以及随后的自体移植）可以提供一种“安全”的方法， 保护”卵巢储备免受疾病病理学的性腺毒性影响，这种方法已经被 自十多年前的早期尝试以来，在癌症患者中使用该技术至少产生了100例活产。 虽然这种方法是有效的，但冷冻保存的卵巢组织的活力和生产力是有限的。 在急性期，移植物内的炎性、缺血性环境严重破坏 移植后 该提议利用了修饰的mRNA的有效生物活性和直接递送 因此，本发明提供了一种将modRNA（modRNA）作为改善卵巢组织自体移植物的存活力和输出的治疗方法的方法。 使用我们小组开发的异种移植平台，我们将研究modRNA的潜力：1） 提高为保持生育力而移植的卵巢组织的活力和产量; 2） 维持一个强大的池卵泡在一个静止的原始状态，将作为一个长期的 用于内分泌功能和卵母细胞输出正常化的储库;以及3）证明这种储库的功效。 在高度相关的临床前动物模型恒河猴中采用这种方法。通过这些实验， 为卵巢组织的稳健、可重复和FDA批准的方法奠定基础 冷冻保存/自体移植在慢性疾病的女孩，希望保持其生殖 选项.

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.