Defining the microenvironment that will enable a long-term bioprosthetic ovary transplant

项目摘要/摘要 卵巢含有有限的潜在卵子和产生性激素的细胞，因此，生命-- 保留辐射或化学诱导卵巢内细胞死亡的癌症治疗可能会导致 卵巢功能不全(POI)伴卵巢激素减少和不孕症。选择拥有生物学的 儿童，是癌症幸存者生活质量的关键指标。患有POI的女性将经历共病 与卵巢激素丢失和预期寿命缩短有关。保持生育能力的唯一方法 对于还没有卵子的儿科患者，则是卵巢组织冷冻保存(OTC)。然后这个组织就可以 移植回来恢复生育和荷尔蒙功能。然而，只有20%-30%的移植会导致 活产，平均产生2-5年的荷尔蒙恢复，使许多人没有生物学上的 儿童和癌症后几十年的生存没有必要的激素产生。一位主要的贡献者 移植卵巢组织的功能缩短是卵巢储备激活的显著高峰 (原始卵泡)和随后的衰竭，至少部分是由于微环境的破坏。 此外，一些患者卵巢内有转移性疾病，因此无法在卵巢内使用该组织。 当前表单。因此，一个安全、长期的生育和荷尔蒙恢复解决方案将包括隔离 卵巢细胞对潜在的癌细胞的功能是必不可少的，并将它们储存在 维持潜在卵库并延长荷尔蒙产生的微环境。我们有 之前开发了一种经过改造的3D打印支架，可以恢复小鼠的生育能力和激素功能。我们 将检验这样一种假设，即母体施加生化和物理线索来控制原始卵泡 激活。此外，我们预测基质细胞的贡献对于完全的卵泡发生是必要的。我们 将使用牛卵巢作为人类卵巢的单排卵模型。在目标1中，我们将研究生物化学 母体的信号及其如何通过定义母体蛋白调节原始卵泡的激活 存在于卵巢中，并调节可能是控制原始卵泡的关键候选蛋白 通过细胞外抑制或诱导细胞内途径激活。在目标2中，我们将调查 卵巢的物理特性，以及它们如何通过定义天然的 卵巢微环境的僵硬和监测原始卵泡对不同物理状态的反应 文化中的属性。在目标3中，我们将研究基质细胞在卵泡发生中的作用。 可移植的支架。我们将定义基质分泌的母体蛋白和旁分泌因子。 细胞，并研究它们在3D打印生物修复支架中对卵泡发生的作用 移植模型。这项申请的目标将建立在我们之前的成功和生物假体卵巢的基础上， 根据这里的结果，这将改善女性目前的生育和荷尔蒙恢复选择。

PROJECT SUMMARY/ABSTRACT The ovaries contain a finite resource of potential eggs and sex hormone-producing cells, and therefore, life- saving cancer treatments that irradiate or chemically induce cell death within the ovaries will likely result in premature ovarian insufficiency (POI) with reduced ovarian hormones and infertility. The option to have biological children, is a key quality of life measure for cancer survivors. Women with POI will experience co-morbidities associated with loss of ovarian hormones and a shorter life expectancy. The only method for fertility preservation for pediatric patients, who do not yet make eggs, is ovarian tissue cryopreservation (OTC). This tissue can then be transplanted back to restore fertility and hormone function. However, only 20 – 30% of transplants result in livebirth and it produces an average of 2 – 5 years of hormone restoration, leaving many without biological children and decades of post-cancer survival without essential hormone production. One major contributor to the shortened function of transplanted ovarian tissue is the significant spike in activation of the ovarian reserve (primordial follicles) and subsequent depletion, at least in part, due to the disruption in the microenvironment. Additionally, some patients have metastatic disease within the ovary and therefore cannot use that tissue in its current form. Therefore, a safe, long-term solution for fertility and hormone restoration would involve isolating the ovarian cells that are essential for function from potential cancer cells and housing them in a microenvironment that maintains the bank of potential eggs and prolongs hormone production. We have previously developed an engineered 3D printed scaffold that restored fertility and hormone function in mice. We will test the hypothesis that the matrisome imposes biochemical and physical cues that controls primordial follicle activation. Additionally, we predict that the contribution of stromal cells is necessary for full folliculogenesis. We will use cow ovaries as mono-ovulatory models of human ovaries. In Aim 1, we will investigate the biochemical cues of the matrisome and how they regulate primordial follicle activation, by defining the matrisome proteins that exist in the ovary and modulating candidate proteins that may be key to controlling primordial follicle activation through extracellular inhibition or induction of intracellular pathways. In Aim 2, we will investigate the physical properties of the ovary and how they control primordial follicle quiescence, by defining the native stiffness of the ovarian microenvironment and monitoring how primordial follicles response to different physical properties in culture. In Aim 3, we will investigate the role of stromal cells in folliculogenesis supported by a transplantable scaffold. We will define the matrisome proteins and paracrine factors that are secreted by stromal cells and investigate how they contribute to folliculogenesis within a 3D printed bioprosthetic scaffold in a transplant model. The aims in this application will build on our previous successes and a bioprosthetic ovary, defined by the results here, would improve current options for fertility and hormone restoration for women.