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.

项目概要 妊娠前三个月切除黄体会导致妊娠终止。 不孕率和约 25% 的妊娠在妊娠早期结束，黄体生理学研究日益增多 黄体功能的破坏是由炎症细胞因子和脂质介质介导的。 介质前列腺素 F2α (PGF2α) 在人类和非人类灵长类动物的腺体中产生，通过 家养农场动物的非妊娠子宫内膜是血管化程度最高的子宫内膜之一。 排卵后黄体血管系统的发育对于成人体内的器官的产生至关重要。 黄体血管系统是独特的，因为它可能是唯一完全血管生成的情况。 成人的血管退化 当黄体在非生育周期结束时退化时， 微血管内皮细胞是最先死亡的细胞，但是这些细胞不表达受体。 黄体分解信号，PGF2α 仅来自类固醇生成大黄体细胞 (LLC)，其源自颗粒细胞。 排卵的卵泡在黄体中拥有这种受体，因此，内皮细胞会发生死亡反应。 在退化的早期阶段，可能是由大黄体细胞响应于 PGF2α。我们的首要假设是黄体退化过程中内皮细胞网络的破坏 是由于大黄体细胞响应 PGF2α 产生的因子所致。本研究的目标是：(1) 确定。 体内诱导黄体退化过程中基因转录的细胞特异性时间变化，以及（2）进行 利用新型 3D 类器官培养物进行机制研究以确定黄体血管消退的介质 我们将使用单核 RNA 测序 (snRNA-seq) 来确定细胞类型的特定时间。 黄体退化过程中基因表达的模式假设：大类固醇黄体细胞对 PGF2α 通过快速增加炎症介质和抗血管生成因子的产生，然后 我们最近开发了在内皮细胞中诱导导致血管破坏和细胞死亡的基因。 新颖的 3D 黄体类器官模型使我们能够确定哪些大黄体细胞分泌产物影响 黄体内皮结构的退化假设：LLC 产生的炎症介质会破坏。 使用 snRNA-seq 分析的最先进方法将提供内皮网络和改变细胞命运。 该研究将建立第一个关于黄体中细胞类型特异性基因表达的无偏转录组图谱。 为特定黄体细胞类型在此过程中基因表达的时间变化提供新的见解 黄体退化的成功完成将填补有关黄体退化的知识空白。 维持性腺功能的关键途径并确定脂质介质的新机制 PGF2α 会破坏性腺功能，深入了解黄体血管退化将为识别提供基础。 维持卵巢和其他生殖组织中脉管系统结构的机制。