Mechanisms underlying sex-dependent pregnancy outcomes caused by fetal and maternal genomic instability

不良妊娠结局可能是多种因素缺陷的结果，例如胎儿或母亲 遗传、环境暴露、子宫功能障碍、先兆子痫、营养、感染、炎症和 胎盘功能不全。从滋养外胚层前体发育出健康的胎盘能够使胎盘正常发育 胎儿和母亲之间的营养物质、气体和废物交换。此外，母体和胎盘固有的 必须控制界面炎症以保护胎儿。该项目解决了基因组如何 卵子发生和胚胎发生过程中的不稳定性（GIN）可能导致性别二态性妊娠结局， 以及胎盘如何特别容易受到这种情况的影响。对这一被低估的研究 不良妊娠结局的原因是由于发现雌性小鼠胚胎显着 当携带某些导致 GIN 的 DNA 复制或修复基因突变时，更容易致命。合而为一 该项目将使用这种模型，女性偏向的致死率是由于易感性增加 炎症，而男性胚胎则受到睾酮的抗炎作用的保护。 初步数据表明胎盘是导致胚胎死亡的敏感组织。值得注意的是， 只有当母鼠也具有 GIN 基因型时，才会发生这种性别偏向的致死率。该项目的目标是 了解驱动杜松子酒诱导致命炎症的组织、细胞和机制。这将是 利用小鼠遗传学、基因组学和胚胎操作的力量来完成。目标 1 将测试是否 胎盘、胚胎或两者都是造成女性致死率的原因。我们假设高度 多倍体滋养层巨细胞可能对 DNA 复制受损和 GIN 特别敏感， 引发先天性炎症。目标 2 解决了为什么卵母细胞必须来自高 GIN 母亲才能进行性别 偏向发生在携带突变致死基因型的胎儿中。初步实验表明，此类水坝 产生线粒体受损的卵母细胞，这一假设将使用线粒体进行测试 增强和组学分析。目标 3 寻求确定致死胚胎的分子基础 炎症，重点关注先天免疫的触发因素。遗传和分子检测的结合将 用于检验核 GIN 导致线粒体 RNA 和 DNA 泄漏，激活 刺激炎症驱动干扰素基因转录的途径。 总的来说，如果成功，结果将有助于解释和解决复发性个案 妊娠丢失可能是妊娠期间内在炎症的基础，并提供了见解 导致不良妊娠结局（包括流产和宫内妊娠）的机制未被充分认识 生长迟缓。

Adverse pregnancy outcomes can be the consequence of defects in several factors, such fetal or maternal genetics, environmental exposures, uterine dysfunction, preeclampsia, nutrition, infection, inflammation, and placental insufficiency. Development of a healthy placenta from trophectoderm precursors enables proper nutrient, gas and waste exchange between the fetus and mother. Additionally, maternal and placenta-intrinsic inflammation at the interface must be controlled to protect the fetus. This project addresses how genomic instability (GIN) during oogenesis and embryogenesis can cause sexually dimorphic pregnancy outcomes, and how the placenta may be especially susceptible to this condition. Research into this underappreciated cause of adverse pregnancy outcomes is motivated by findings that female mouse embryos are dramatically more prone to lethality when bearing certain GIN-causing mutations of DNA replication or repair genes. In one such model that will be utilized in this project, the female-biased lethality was due to increased susceptibility to inflammation, whereas male embryos were protected by the anti-inflammatory effects of testosterone. Preliminary data implicate the placenta as the sensitive tissue underlying the embryonic death. Remarkably, this sex-biased lethality occurred only if the dam also had a GIN genotype. The goals of this project are to understand the tissue(s), cells, and mechanisms driving GIN-induced lethal inflammation. This will be accomplished using the power of mouse genetics, genomics, and embryo manipulation. Aim 1 will test whether the placenta, the embryo, or both, are responsible for female-biased lethality. We hypothesize that the highly polyploid trophoblast giant cells may be especially sensitive to compromised DNA replication and GIN, triggering innate inflammation. Aim 2 addresses why oocytes must come from high GIN mothers for the sex bias to occur in fetuses bearing the mutant lethal genotype. Preliminary experiments implicate that such dams produce oocytes with compromised mitochondria, and this hypothesis will be tested using mitochondrial augmentation and -omics analyses. Aim 3 seeks to identify the molecular basis of lethal embryonic inflammation, with a focus on triggers of innate immunity. A combination of genetic and molecular assays will be used to test the hypothesis that nuclear GIN leads to mitochondrial RNA and DNA leakage, activating a pathway(s) that stimulates transcription of inflammation-driving interferon genes. Overall, if successful, the results will be relevant for interpreting and addressing individual cases of recurrent pregnancy loss that may have a basis in intrinsic inflammation during gestation, and provide insights into underappreciated mechanisms causing adverse pregnancy outcomes including miscarriage and intrauterine growth retardation.