Characterization & Prevention of Chemotherapy-Induced Damage to Ovarian Reserve

描述（由申请人提供）：癌症治疗引起的卵巢功能衰竭是一个重大的公共卫生问题，可能有1%的人口在生殖寿命期间受到影响。我们的总体目标是了解和防止这种治疗造成的损害。在上一个资助期，我们取得了重大发现，为下一个资助期奠定了基调。我们发现，促性腺毒性化疗主要通过引起双链（DSB） DNA断裂导致原始卵泡死亡，作为对这种损伤的反应，卵母细胞会产生ATM介导的DNA DSB修复反应（Aging, 2011）。这种反应可能使一些原始卵泡在化疗中存活下来。此外，我们发现DNA DSB修复反应在卵母细胞减轻基因毒性损伤和衰老的方式中至关重要，并且DNA DSB修复缺陷的女性，特别是BRCA1突变携带者，可能容易过早耗尽卵巢储备（Science Translational Medicine, 2013）。此外，我们发现S1P，一种天然存在的神经酰胺死亡途径抑制剂，可以减少化疗诱导的人类卵巢异种移植的原始卵泡死亡（human Reproduction 2014）。由于这些发现和DNA DSB修复反应是统一的主题，我们更新应用的具体目标是：1。为了确定brca突变携带者是否更容易因其固有的DNA DSB修复缺陷而导致化疗诱导的卵巢卵泡丢失；2. 揭示一些原始卵泡能够在化疗损伤中存活的机制；3. 了解S1P对人类卵巢原始卵泡池的保护作用。为了实现这些目标，我们将利用临床研究，单细胞实时PCR以及微阵列策略，原位杂交，基因干扰，结合人类卵巢异种移植和激光捕获方法。虽然这些研究主要是翻译性的，并将利用人类材料，但小鼠模型也将用于加强机制工作。

DESCRIPTION (provided by applicant): Cancer therapy-induced ovarian failure is a significant public health problem with potentially 1% of population being affected over reproductive life span. Our overall goal is to understand and prevent the damage caused by such treatments. In the previous grant period we made significant discoveries which set the tone for the next grant period. We discovered that gonadotoxic chemotherapeutics result in primordial follicle death primarily by causing double strand (DSB) DNA breaks, and in response to this insult, oocytes mount an ATM- mediated DNA DSB repair response (Aging, 2011). This response may enable some primordial follicles to survive chemotherapy. Furthermore, we found that DNA DSB repair response is critical in the way oocytes mitigate genotoxic insult and aging, and that women who are deficient in DNA DSB repair, specifically BRCA1- mutation carriers, maybe prone to prematurely depleting their ovarian reserve (Science Translational Medicine, 2013). In addition, we showed that S1P, a naturally occurring ceramide death-pathway inhibitor, reduces chemotherapy-induced primordial follicle death in human ovarian xenografts (Human Reproduction 2014). Stemming from these revelations and the DNA DSB repair response being the unifying theme, our specific aims for the renewal application are: 1. To determine if BRCA-mutation carriers are more prone to chemotherapy-induced ovarian follicle loss because of their inherent deficiency of DNA DSB repair; 2. To reveal the mechanisms by which some primordial follicles are able to survive chemotherapy insult; 3. To understand how S1P protects human ovarian primordial follicle pool. To achieve these aims we will utilize clinical studies, single cel real time PCR as well as microarray strategies, in situ hybridization, gene interference, coupled with human ovarian xenografting and laser capture approaches. While the studies are primarily translational and will utilize human material, mouse models will also be used to strengthen the mechanistic work.