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.

项目概要 扩张和刮宫术 (D&C) 是全世界对女性最常见的外科手术之一 世界上大约 40% 接受 D&C 的女性会出现宫腔粘连 (iUA) 或 Asherman。 产后（出生后 6 周期间），但在非孕妇 D&C 后非常罕见 (<1%)，原因如下 这是一种以子宫内纤维化和疤痕为特征的衰弱性疾病。 患有 iUA 的人患有不孕症、反复流产和各种危险的妊娠并发症 （例如早产）而子宫内膜间充质干细胞/祖细胞（eMPC）对于子宫内膜至关重要。 修复、这些细胞的作用以及产后易感性的潜在分子机制 子宫内膜纤维化和异常修复尚不清楚，本申请特别侧重于定义。 eMPC 及其基质成纤维细胞后代在异常子宫修复中发挥的作用核心假设。 是最近产后子宫的 eMPC 更加衰老，并且它们对 与非妊娠子宫的 eMPC 相比，子宫损伤，导致纤维化和疤痕形成。 方法是使用我们新型的产后小鼠子宫损伤模型，该模型概括了子宫损伤的易感性 利用它，我们将定义 eMPC 的动态变化及其差异。 产后与非妊娠时对子宫损伤的反应，使用谱系追踪 eMPC 子集进行识别 成为促纤维化成纤维细胞，并有条件地消融每个 eMPC 子集以定义其 在子宫内膜纤维化中的功能作用（目标1）我们将从女性身上获得新鲜的人类子宫内膜组织。 进行产后 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，以详细了解导致疾病发生的细胞和分子差异 人类子宫内膜形成iUA（目标2）在目标3中，我们将获得当时存档的子宫内膜标本。 发展阿什曼与非阿什曼的女性煽动 D&C 事件的比较 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 将其与免疫荧光相结合，以获得有关 eMPC 及其细胞的详细分子见解 组织内的相互作用，确定预防 iUA 的新治疗靶点。 在概念和技术上都具有创新性，并且共同将通过填补这一领域对该领域产生广泛的影响 我们使用组学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 方法，这是 NICHD 生育和不孕不育科的主要研究重点。 最终，从该提案中获得的知识不仅对于我们理解许多问题具有无价的价值 异常子宫内膜修复的更微妙的条件，但提供了对身体的独特见解 生理性抗纤维化伤口愈合机制使人们更深入地了解伤口的发病机制 其他器官致命的特发性纤维化疾病。