Vascular remodeling in the ovary

卵巢血管重塑

• 批准号: 10724873
• 负责人: JOHN S DAVIS
• 金额: $ 15.35万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724873
• 关键词: 3-Dimensional; Adult; Affect; Angiogenesis Inhibitors; Angiogenic Factor; Atlases; Blood Vessels; CDH5 gene; Cattle; Cell Death; Cells; Conditioned Culture Media; Cytokine Gene; Cytokine Receptors; Data; Development; Endometrium; Endothelial Cells; Endothelium; Excision; Fertility; First Pregnancy Trimester; Foundations; Freezing; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene set enrichment analysis; Genes; Genetic Transcription; Gland; Goals; Granulosa-Lutein Cells; Human; In Vitro; Infertility; Inflammation Mediators; Inflammatory; Intramuscular Injections; Knowledge; Livestock; Luteal Cells; Luteal Phase; Luteolysis; Mediating; Mediator; Modeling; Nuclear RNA; Organ; Organoids; Ovarian; Ovary; Ovulation; Pathway interactions; Peptide Initiation Factors; Physiology; Pregnancy; Pregnancy Maintenance; Process; Production; Progesterone; Prostaglandins; Reporting; Research; Saline; Secretory Cell; Signal Transduction; Structure; System; Termination of pregnancy; Testing; Tissues; Up-Regulation; Uterus; Vascular Endothelial Cell; Vascular remodeling; Vascularization; angiogenesis; cell type; corpus luteum; cytokine; differential expression; gene induction; gene repression; genetic signature; gonad function; granulosa cell; in vitro Model; in vivo; insight; lipid mediator; nonhuman primate; novel; receptor; release factor; reproductive; response; single nucleus RNA-sequencing; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Corpus luteum removal during the first trimester of pregnancy leads to pregnancy termination. With rising infertility rates and ~25% of pregnancies ending in the first trimester, research in luteal physiology is increasingly relevant. Disruption of luteal function is mediated by inflammatory cytokines and lipid mediators. The key lipid mediator prostaglandin F2α (PGF2α) is produced within the gland in humans and non-human primates and by the non-pregnant endometrium in domestic farm animals. The corpus luteum is one of the most vascularized organs in the adult body. Development of the luteal vasculature following ovulation is vital for the production of progesterone. The luteal vasculature is unique because it is possibly the only case of full angiogenesis and angioregression in the adult. When the corpus luteum regresses at the end of a non-fertile reproductive cycle, microvascular endothelial cells are the first cells to die. However, these cells do not express receptors for the luteolytic signal, PGF2α. Only the steroidogenic large luteal cells (LLC), which derive from the granulosa cells of the ovulated follicle, possess this receptor in the corpus luteum. Therefore, the endothelial cell death response during the early stages of regression is likely initiated by factors released by large luteal cells in response to PGF2α. Our overarching hypothesis is that the disruption of endothelial cell networks during luteal regression are due to factors produced by large luteal cells in response to PGF2α. The goals of this study are to: (1) identify cell-specific temporal changes in gene transcription during induced luteal regression in vivo, and (2) conduct mechanistic studies to determine mediators of luteal angioregression utilizing a novel 3D organoid culture system. We will use single nuclear RNA sequencing (snRNA-seq) to determine the cell-type specific temporal patterns of gene expression during luteal regression. Hypothesis: Large steroidogenic luteal cells respond to PGF2α by rapidly increasing production of inflammatory mediators and anti-angiogenic factors, followed by induction of genes in endothelial cells that contribute to vascular disruption and cell death. We recently developed novel 3D luteal organoid model to allow us to determine which large luteal cell secretory product(s) affect the regression of luteal endothelial structures. Hypothesis: inflammatory mediators produced by LLC will disrupt endothelial networks and alter cell fate. The state-of-the-art approach using snRNA-seq profiling will provide the first unbiased transcriptomic atlas of the cell type specific gene expression in the corpus luteum. The studies will provide novel insight into the temporal changes in gene expression in specific luteal cell types during the process of luteal regression. Successful completion of the proposed research will fill a gap in knowledge about the pathways crucial for maintaining gonadal function and identify novel mechanisms by which the lipid mediator PGF2α disrupts gonadal function. Insight into luteal angioregression will provide the foundations for identifying mechanisms to maintain vasculature structures in the ovary and other reproductive tissues.

项目摘要 在妊娠的第一个三个月期间切除黄体会导致妊娠终止。上涨的 不孕率和~25%的妊娠在妊娠早期结束，黄体生理学的研究越来越多， 相关的黄体功能的破坏是由炎性细胞因子和脂质介质介导的。关键脂质 介质前列腺素F2α（PGF 2 α）在人类和非人类灵长类动物的腺体内产生， 家畜未怀孕的子宫内膜。黄体是血管化程度最高的 成人体内的器官。排卵后黄体脉管系统的发育对于产生 孕酮黄体脉管系统是独特的，因为它可能是唯一的完全血管生成的情况， 成人血管退行。当黄体在非生育性生殖周期结束时退化时， 微血管内皮细胞是首先死亡的细胞。然而，这些细胞不表达受体， 黄体溶解信号，PGF 2 α。只有产类固醇的大黄体细胞（LLC），它来自于卵巢颗粒细胞， 排卵的卵泡在黄体中具有这种受体。因此，内皮细胞死亡反应 在退化的早期阶段，可能是由大黄体细胞释放的因子引起的， PGF 2 α。我们的总体假设是，黄体退化过程中内皮细胞网络的破坏 是由于大的黄体细胞对PGF 2 α的反应而产生的因子。本研究的目的是：（1）识别 在体内诱导的黄体退化期间基因转录的细胞特异性时间变化，和（2）进行 利用新型3D类器官培养确定黄体血管退化介质的机制研究 系统我们将使用单核RNA测序（snRNA-seq）来确定细胞类型特异性时间序列。 黄体退化过程中的基因表达模式。假设：大的类固醇生成黄体细胞响应于 PGF 2 α通过迅速增加炎症介质和抗血管生成因子的产生， 诱导内皮细胞中导致血管破裂和细胞死亡的基因。我们最近开发 一种新的3D黄体类器官模型，使我们能够确定哪些大的黄体细胞分泌产物影响了 黄体内皮结构退化。假设：LLC产生的炎症介质会破坏 内皮网络和改变细胞命运。使用snRNA-seq分析的最先进方法将提供 黄体中细胞类型特异性基因表达的第一个无偏转录组图谱。这些研究将 提供了新的见解，在特定的黄体细胞类型的基因表达的时间变化的过程中， 黄体退化成功完成拟议的研究将填补知识的空白， 维持性腺功能的关键途径，并确定脂质介质 PGF 2 α破坏性腺功能。对黄体血管退行性病变的深入了解将为识别 维持卵巢和其他生殖组织中脉管系统结构的机制。