THE ROLE OF MESENCHYMAL PROGENITOR CELLS IN ABNORMAL UTERINE REPAIR

间充质祖细胞在异常子宫修复中的作用

• 批准号: 10660189
• 负责人: Reshef Tal
• 金额: $ 55.08万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660189
• 关键词: Ablation; Adhesions; Adult; Affect; Archives; Asherman Syndrome; Bar Codes; Biology; Birth; Cell Aging; Cell Communication; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Cicatrix; Cre-LoxP; Dangerousness; Data; Development; Dilatation and Curettage; Disease; Endometrial; Endometrium; Event; Fertility; Fibroblasts; Fibrosis; Functional disorder; Gene Expression Profile; Genetic; Goals; Habitual Abortion; Heart; Human; Immunofluorescence Immunologic; Immunohistochemistry; In Vitro; Infertility; Injury; Knowledge; Liver; Lung; Mechanics; Medical; Menstrual cycle; Mesenchymal Stem Cells; Modeling; Molecular; Mus; National Institute of Child Health and Human Development; Natural regeneration; Operative Surgical Procedures; Organ; Pathogenesis; Pathway interactions; Patients; Physiological; Play; Population; Postpartum Period; Predisposition; Pregnancy; Pregnancy Complications; Pregnancy loss; Premature Birth; Prevention; Property; Proteins; Recurrence; Reporter; Research Priority; Role; Specimen; System; Time; Tissues; Trauma; Uterus; Woman; Work; cell type; exhaustion; iatrogenic injury; idiopathic pulmonary fibrosis; in vitro Assay; innovation; insight; mouse model; multiple omics; new therapeutic target; novel; prevent; profibrotic fibroblast; repaired; reproductive; response; response to injury; senescence; sequencing platform; single cell technology; single-cell RNA sequencing; stem cells; stemness; tissue mapping; transcriptome; wound healing

PROJECT SUMMARY Dilation & curettage (D&C) is one of the most common surgical procedures performed on women throughout the world. Intrauterine adhesions (iUA), or Asherman, develop in about 40% of women who undergo D&C in the postpartum (6 week period after birth), but is very rare after D&C in nonpregnant women (<1%) for reasons that are not well understood. It is a debilitating condition characterized by intrauterine fibrosis and scarring. Patients with iUA suffer from infertility, recurrent pregnancy loss and a broad range of dangerous pregnancy complications (e.g. preterm birth). While endometrial mesenchymal stem/progenitor cells (eMPCs) are crucial for endometrial repair, the role of these cells and underlying molecular mechanisms in this postpartum susceptibility of the endometrium to fibrosis and abnormal repair are unknown. This application is specifically focused on defining the role that eMPCs and their stromal fibroblast progeny play in abnormal uterine repair. The central hypothesis is that eMPCs of the recently postpartum uterus are more senescent and inherently different in their response to uterine injury compared to eMPCs of the nonpregnant uterus, leading to fibrosis and scar formation. The approach is to use our novel postpartum mouse uterine injury model which recapitulates the susceptibility of the human postpartum uterus to injury. Using it, we will define the dynamic changes in eMPCs and their differential response to uterine injury in the postpartum vs. nonpregnant, identify using lineage tracing the eMPC subsets that become the profibrotic fibroblast cells, and conditionally ablate each of these eMPC subsets to define their functional role in endometrial fibrosis (Aim 1). We will obtain fresh human endometrial tissue from women undergoing postpartum D&C and compare it to nonpregnant tissue using innovative single cell technology and functional in vitro studies to gain detailed insights into the cellular and molecular differences that predispose the human endometrium to form iUA (Aim 2). In Aim 3, we will obtain archived endometrial specimens from the time of inciting D&C event from women who developed Asherman vs. non-Asherman. We will use the innovative deterministic barcoding in tissue spatial multi-omics sequencing (DBiT-seq) platform and integrate it with immunofluorescence to gain detailed molecular insights regarding eMPCs and their cell interactions within the tissue, identifying novel therapeutic targets for iUA prevention. The proposed aims are conceptually and technically innovative and together will have a broad impact on the field by filling a substantial gap in our fundamental knowledge of endometrial biology and infertility pathogenesis using Omics approach, which are major research priorities of the Fertility and Infertility Branch of the NICHD. Ultimately, the knowledge gained from this proposal will not only be invaluable to our understanding of many more subtle conditions of abnormal endometrial repair, but provide unique insights into the body’s physiological anti-fibrotic wound healing mechanisms leading to a deeper understanding of the pathogenesis of fatal idiopathic fibrotic diseases in other organs.

项目摘要 扩张和刮宫术（D&C）是整个世界对女性进行的最常见的外科手术之一。 世界子宫内粘连（iUA），或Asherman，发生在约40%的妇女谁接受D&C在世界各地。 产后（出生后6周），但在非妊娠妇女中D&C后非常罕见（<1%），原因是 并没有得到很好的理解。这是一种使人衰弱的疾病，其特征是宫内纤维化和疤痕。患者 iUA患者患有不孕症、反复流产和各种危险的妊娠并发症 (e.g.早产）。虽然子宫内膜间充质干/祖细胞（eMPCs）在子宫内膜异位症中至关重要， 修复，这些细胞的作用和潜在的分子机制，在这种产后易感性的 子宫内膜纤维化和异常修复是未知的。此应用程序专门用于定义 eMPC及其基质成纤维细胞后代在异常子宫修复中的作用。核心假设 最近产后子宫的eMPCs更衰老，并且在它们对 子宫损伤，导致纤维化和瘢痕形成。的 方法是使用我们的新的产后小鼠子宫损伤模型，该模型概括了 人体产后子宫损伤。使用它，我们将定义eMPC的动态变化及其差异 产后与非妊娠期对子宫损伤的反应，使用谱系追踪eMPC子集进行鉴定 成为促纤维化成纤维细胞，并有条件地消融这些eMPC亚群中的每一个，以确定它们的 在子宫内膜纤维化中的作用（目的1）。我们将从女性身上获取新鲜的人类子宫内膜组织 接受产后D&C，并使用创新的单细胞技术将其与非妊娠组织进行比较， 功能性体外研究，以详细了解易患糖尿病的细胞和分子差异， 人子宫内膜形成iUA（Aim 2）。在目标3中，我们将获得存档的子宫内膜标本， 激发D&C事件的女性谁开发了Asherman与非Asherman。我们将使用 组织空间多组学测序（DBiT-seq）平台中的创新确定性条形码 将其与免疫荧光结合，以获得关于eMPC及其细胞详细分子见解 组织内的相互作用，识别用于iUA预防的新的治疗靶点。拟议目标 在概念和技术上都是创新的，它们将共同对该领域产生广泛影响， 使用组学我们对子宫内膜生物学和不孕症发病机制的基础知识存在巨大差距 这是NICHD生育和不育分支的主要研究重点。 最终，从这一建议中获得的知识不仅对我们了解许多 子宫内膜修复异常的更微妙的条件，但提供了独特的见解，身体的 生理抗纤维化伤口愈合机制，从而更深入地了解 其他器官的致命性特发性纤维化疾病。