The Impact of Leukocyte-Derived OSM on Ovulation and Luteinization in the Human Ovary

白细胞衍生的 OSM 对人类卵巢排卵和黄体化的影响

• 批准号: 10684327
• 负责人: Yohan Choi
• 金额: $ 7.65万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10684327
• 关键词: Acute; Affect; Binding; Bioinformatics; Biological; Cell Culture Techniques; Cells; Data; Dinoprostone; EGF gene; Family; Fertility; Fertilization in Vitro; Gene Expression; Genes; Gonadotropins; Human; Human Chorionic Gonadotropin; IL6 gene; Infertility; Inflammation; Inflammatory Response; Interleukin 6 Receptor; Knowledge; Leukocytes; Luteal Cells; Luteinization; MAPK3 gene; Mediator; Messenger RNA; OSMR gene; Ovary; Ovulation; PTGS2 gene; Pathway interactions; Phase; Procedures; Process; Production; Progesterone; Progesterone Receptors; Proteins; Recombinants; Role; Sampling; Signal Pathway; Signal Transduction; Signaling Molecule; Stat3 Signaling Pathway; Supplementation; Testing; Time; Tissues; Transducers; Up-Regulation; Woman; aspirate; design; differential expression; female fertility; fertility improvement; granulosa cell; improved; in vivo; leukemia inhibitory factor receptor; male; novel; oncostatin M; ovarian dysfunction; proliferative phase Menstrual cycle; receptor; receptor expression; recruit; single-cell RNA sequencing; spatiotemporal; transcriptome; transcriptome sequencing

ABSTRACTS The ovulatory process is the mid-cycle gonadotropin surge-induced acute inflammatory response. Leukocytes are critical effectors in the inflammatory response. However, the mechanisms of leukocyte recruitment and the specific roles of leukocytes in the ovulatory follicle remain elusive, especially in humans. The current study proposes to elucidate how the mid-cycle LH surge coordinates leukocyte recruitment and the expression of genes in preovulatory follicular cells to bring about successful ovulation and luteinization in the human ovary. From our pilot single-cell transcriptome analyses using human follicular aspirates from women undergoing in vitro fertilization, we discovered that OSM (oncostatin-M) is exclusively expressed in specific types of leukocytes, whereas its receptors, OSMR and LIFR (leukemia inhibitory factor receptor) and their co- receptor, IL6ST (IL6 family signal transducer) are present predominantly in granulosa cells. Similarly, in granulosa cells of dominant follicles collected from normally cycling women throughout the periovulatory period, we found that OSMR mRNA levels were markedly increased after hCG stimulation. In addition, our preliminary data showed that hCG increased the expression of OSM receptors, OSMR and IL6ST in primary human granulosa cell cultures, mimicking in vivo up-regulation of OSMR. Our preliminary data also revealed that OSM could activate specific signaling pathways and stimulate progesterone and prostaglandin E2 production, two crucial mediators of ovulation. Based on these novel data, we hypothesized that 1) the LH surge induces the recruitment of OSM- expressing leukocytes, while increasing the expression of its receptors (OSMR, LIFR, IL6ST) in follicular cells of preovulatory follicles, and 2) leukocyte-derived OSM, upon binding to its receptors in preovulatory follicular cells, activates specific signaling pathways, and regulates the expression of specific ovulatory genes, thereby promoting necessary ovulatory changes. These hypotheses will be tested by first characterizing expression profiles of OSM in leukocytes and its receptors, OSMR, LIFR, and IL6ST in follicular cells of the dominant follicles collected throughout the periovulatory period and examining the regulatory mechanisms involved in the expression of OSM receptors in human granulosa cells (Specific Aim 1). Next, we will delineate OSM- activated intracellular signaling pathways in human granulosa cells (Specific Aim 2). Lastly, we will elucidate the impact of leukocyte-derived OSM on the periovulatory process by identifying OSM-regulated downstream genes (RNA-seq analysis) and evaluating cellular/biological impacts on affected pathways using human granulosa cell cultures. The proposed study will demonstrate “for the first time” OSM as a novel ovulatory factor of leukocyte-origin and identify its specific roles in the human ovary, thus filling the gap in our understahopteanding of the role of leukocytes in the ovulatory process.

摘要 排卵过程是周期中促性腺激素激增引起的急性炎症反应。 白细胞是炎症反应的关键效应物，然而，白细胞的机制。 排卵卵泡中白细胞的募集和具体作用仍然难以捉摸，尤其是在人类中。 目前的研究旨在阐明周期中期 LH 激增如何协调白细胞募集和 排卵前卵泡细胞中的基因表达，以实现成功排卵和黄体化 人类卵巢。 来自我们使用女性卵泡抽吸物进行的试点单细胞转录组分析 在体外受精过程中，我们发现 OSM（制瘤素-M）仅在特定的细胞中表达。 白细胞的类型，而其受体 OSMR 和 LIFR（白血病抑制因子受体）及其共同 类似地，IL6ST（IL6家族信号转导器）受体主要存在于颗粒细胞中。 从正常周期女性的整个排卵期收集优势卵泡的颗粒细胞 期间，我们发现 hCG 刺激后 OSMR mRNA 水平显着增加。 初步数据显示，hCG 增加了原发性小鼠中 OSM 受体、OSMR 和 IL6ST 的表达。 我们的初步数据还揭示了人颗粒细胞培养物，模拟体内 OSMR 的上调。 OSM 可以激活特定的信号通路并刺激黄体酮和前列腺素 E2 生产，排卵的两个重要介质。 基于这些新数据，我们勇敢地承认 1) LH 激增会诱导 OSM- 表达白细胞，同时增加滤泡细胞中其受体（OSMR、LIFR、IL6ST）的表达 排卵前卵泡的，以及 2) 白细胞衍生的 OSM，在与排卵前卵泡中的受体结合后 细胞，激活特定的信号通路，调节特定排卵基因的表达，从而 促进必要的排卵变化将通过首先表征表达来检验。 白细胞及其受体中的 OSM 谱、优势滤泡细胞中的 OSMR、LIFR 和 IL6ST 在整个排卵期收集卵泡并检查参与的调节机制 OSM 受体在人颗粒细胞中的表达（具体目标 1） 接下来，我们将描述 OSM-。 激活人颗粒细胞的细胞内信号通路（具体目标 2）。 通过识别 OSM 调节的下游，研究白细胞来源的 OSM 对排卵期过程的影响 基因（RNA-seq 分析）并使用人类评估对受影响途径的细胞/生物学影响 拟议的研究将“首次”证明 OSM 作为一种新型排卵剂。 并确定其在人类卵巢中的具体作用，从而填补了我们的空白 了解白细胞在排卵过程中的作用。