Improving Primordial Follicle Survival After Transplantation of Cryopreserved Hum

冷冻保存的蜂移植后提高原始卵泡的存活率

• 批准号: 8113055
• 负责人: KUTLUK H OKTAY
• 金额: $ 24.15万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8113055
• 关键词: Aftercare; Apoptotic; Biological Preservation; Cancer Patient; Cell Death; Ceramides; Cryopreservation; Development; Dyes; Effectiveness; Endocrine; Fertility; Freezing; Goals; Graft Survival; Human; Human Chorionic Gonadotropin; Hypoxia; Imaging Techniques; Infertility; Injection of therapeutic agent; Ischemia; Knowledge; Late Effects; Life; Longevity; Malignant Neoplasms; Minor; Molecular; Morphology; Mus; Oocytes; Outcome; Ovarian; Ovarian Stimulations; Ovarian Tissue; Ovary; Ovulation; Pathway interactions; Patients; Pharmacological Treatment; Phase; Physiological; Premature Ovarian Failure; Primordial Follicle; Procedures; Process; Production; Public Health; Quality of life; Research; Retrieval; Severe Combined Immunodeficiency; Survival Rate; Techniques; Testing; Time; Tissue Survival; Translating; Transplantation; Vascularization; Xenograft Model; Xenograft procedure; base; cancer therapy; chemotherapy; egg; graft function; human tissue; improved; inhibitor/antagonist; innovation; neovascularization; novel strategies; oocyte maturation; research study; response; sphingosine 1-phosphate; steroid hormone; vasculogenesis

DESCRIPTION (provided by applicant): Ovarian tissue cryopreservation is one of the key approaches to fertility preservation. It does not require ovarian stimulation and can be done in a relatively short time and even in minors. While previous research indicated that primordial follicle losses are relatively small after freezing and thawing of ovarian tissue, a large fraction of the ovarian reserve is lost during the initial ischemia after ovarian transplantation. Thus any approach that can enhance and facilitate revascularization of ovarian transplants is likely to improve the outcome of the procedure. Hence, the long-term objective of this project is to develop novel strategies to improve tissue survival after transplantation of ovarian tissue in humans. The overarching hypothesis of our proposal is that the primordial follicle survival and ovarian graft function can be improved by pharmacological treatments enhancing vasculogenesis. This will be studied under the following specific aims: 1. To determine whether a ceramide-induced cell death pathway inhibitor, sphingosine-1 phosphate (S1P), protects against post-transplant ischemia-induced damage to ovarian primordial follicle pool in a human ovarian xenograft model. Recent studies have shown that S1P, an inhibitor of ceramide-induced apoptotic cell death, can enhance physiological vascularization processes. Thus we hypothesize that S1P will enhance primordial follicle survival during the initial ischemic phase after ovarian transplantation by promoting neovascularization of ovarian grafts. To test the effectiveness of S1P in augmenting ovarian transplant neovascularization, we will employ a human ovarian xenograft model based on Severe-Combined- Immunodeficiency (SCID) mice. We will determine whether S1P improves graft survival and neovascularization by primordial follicle survival, histological and molecular assessment of neovascularization, evidence of hypoxia by HIF-1a, as well as by utilization of intravital dye injections and innovative live imaging techniques. 2. To determine whether S1P treatment improves the functionality of frozen-thawed ovarian tissue transplants in a xenograft model. While S1P may enhance vascularization of ovarian transplants and increase survival of primordial follicles, this does not necessarily mean that these grafts will have extended function. Thus, we hypothesize that not only S1P will improve primordial follicle survival in ovarian xenografts but these follicles will be functional. A larger surviving functional primordial follicle pool should translate into extended functionality of ovarian grafts. To show that S1P treatment can thus enhance functionality of ovarian transplants, we will conduct long-term ovarian xenografting experiments. After 16 weeks of xenografting, functionality will be assessed and compared between S1P treated and control grafts by analysis of follicle and oocyte morphology, oocyte maturation and ovulation rates in response to hCG administration, steroid hormone production, as well as parthenogenic activation. PUBLIC HEALTH RELEVANCE: Premature ovarian failure and infertility are significant consequences of cancer treatments with major impact on quality of life and public health. Transplantation of cryopreserved ovarian tissue is one of the emerging key fertility preservation strategies in cancer patients but the technique is limited due to low survival of eggs in ovarian tissue. The overall goal of this project is to enhance ovarian graft survival and function by developing pharmacological treatments that can enhance new vessel development in ovarian transplants.

描述（由申请人提供）：卵巢组织冷冻保存是保存生育能力的关键方法之一。它不需要卵巢刺激，并且可以在相对较短的时间甚至未成年人中完成。虽然先前的研究表明，冷冻和融化卵巢组织后原始卵泡损失相对较小，但在卵巢移植后初始缺血期间，卵巢储备的很大一部分丢失。因此，任何可以增强和促进卵巢移植的方法的方法都可能改善该程序的结果。因此，该项目的长期目标是制定新的策略，以改善人类卵巢组织移植后的组织存活。我们建议的总体假设是，可以通过增强血管生成的药理治疗来改善原始卵泡存活和卵巢移植功能。这将在以下特定目的下进行研究：1。确定神经酰胺诱导的细胞死亡途径抑制剂鞘氨氨酸-1磷酸盐（S1P）是否可以防止移植后缺血诱导的人类卵巢Xenogograft模型中卵巢原始卵泡对卵巢原始卵泡池的损害。最近的研究表明，S1P是神经酰胺诱导的凋亡细胞死亡的抑制剂，可以增强生理血管化过程。因此，我们假设S1P通过促进卵巢移植物的新血管形成，在卵巢移植后的初始缺血阶段增强原始卵泡存活。为了测试S1P在增强卵巢移植新血管形成方面的有效性，我们将采用基于严重混合的免疫缺陷（SCID）小鼠的人类卵巢异种移植模型。我们将确定S1P是否通过原始卵泡存活，组织学和分子评估新生血管形成，HIF-1A缺氧的证据以及利用浸润性染料注射和创新的实时成像技术来改善移植物的存活和新血管化。 2。确定S1P处理是否改善了异种移植模型中冷冻透视卵巢组织移植的功能。尽管S1P可以增强卵巢移植的血管化并增加原始卵泡的存活率，但这并不一定意味着这些移植物将具有扩展功能。因此，我们假设S1P不仅会改善卵巢异种移植物中的原始卵泡存活率，而且这些卵泡将起作用。较大的存活功能原始卵泡池应转化为卵巢移植物的扩展功能。为了表明S1P处理可以增强卵巢移植的功能，我们将进行长期的卵巢异种反应实验。在定异武后16周后，通过分析卵泡和卵母细胞形态，卵母细胞成熟和排卵率，对HCG给药，类固醇激素的产生，以及par源激活，将评估和对照移植物之间的功能和对照移植物进行比较。 公共卫生相关性：过早的卵巢衰竭和不育是癌症治疗的重大后果，对生活质量和公共卫生产生了重大影响。冷冻保存的卵巢组织的移植是癌症患者的新兴关键生育策略之一，但由于卵巢组织中卵的存活较低，该技术受到限制。该项目的总体目标是通过开发可以增强卵巢移植中新血管发展的药理治疗方法来增强卵巢移植物的生存和功能。