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 扩张和凝结（D＆C）是对整个女性进行的最常见的外科手术程序之一 世界。插入粘附（IUA）或Asherman在大约40％的妇女中发展了D＆C 产后（出生后6周），但在非怀孕妇女的D＆C之后非常罕见（<1％） 不太了解。这是一种以宫内纤维化和疤痕为特征的令人衰弱的状况。患者 IUA患有不育，反复怀孕丧失和广泛的危险怀孕并发症 （例如早产）。子宫内膜间质茎/祖细胞（EMPC）对于子宫内膜至关重要 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中 子宫内膜纤维化和异常修复尚不清楚。该应用程序专门针对定义 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。中心假设 是最近产后子宫的EMPC在对其对的反应时更加感官和固有不同 与非怀孕子宫的EMPC相比，子宫损伤导致纤维化和疤痕形成。这 方法是使用我们新颖的产后小鼠子宫损伤模型，该模型概括了 人产后子宫受伤。使用它，我们将定义EMPC及其差异的动态变化 对产后与非怀孕的子宫损伤的反应，使用谱系追踪EMPC子集 这变成了纤维细胞的纤维细胞，并有条件地消除了这些EMPC子集以定义其 子宫内膜纤维化中的功能作用（AIM 1）。我们将从女性那里获得新鲜的人子宫内膜组织 使用创新的单细胞技术和 功能性研究，以获取对细胞和分子差异的详细见解 人子宫内膜形成IUA（AIM 2）。在AIM 3中，我们将从时间开始获得存档的子宫内膜标本 煽动D＆C事件的妇女，这些妇女发展了Asherman vs.非Asherman。我们将使用 在组织空间多摩学测序（DBIT-SEQ）平台和组织空间中的创新确定性条形码和 将其与免疫荧光整合，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，鉴定了预防IUA的新型治疗靶标。拟议的目标 在概念上和技术上都是创新的，并且通过填充一个 我们对使用OMICS的子宫内膜生物学和不孕发病机理的基本知识的巨大差距 方法是NICHD生育和不育分支的主要研究优先事项。 最终，从这一提议中获得的知识不仅对我们对许多人的理解非常宝贵 异常子宫内膜修复的更微妙的条件，但为人体提供了独特的见解 生理抗纤维化伤口愈合机制，从而更深入地了解 其他器官中致命的特发性纤维化疾病。