Neutrophils and Monocytes in Pediatric Ischemic Stroke

小儿缺血性中风中的中性粒细胞和单核细胞

• 批准号: 10629365
• 负责人: Chia-Yi Kuan
• 金额: $ 43.09万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629365
• 关键词: 1 year old; Acute; Adaptive Immune System; Address; Adult; Affect; Birth; Blood - brain barrier anatomy; Caring; Cells; Cerebrovascular system; Childhood; Childhood stroke; Clinical Management; Cognitive deficits; Crime; Data; Development; Dichloromethylene Diphosphonate; Family; Flow Cytometry; Gelatinase B; Gene Expression; Goals; Heterogeneity; Immune; Immune System Diseases; Immune Targeting; Immune response; Immunotherapy; Incidence; Infant; Infarction; Infection; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injections; Interleukin-1; Invaded; Ischemic Stroke; Leptomeninges; Lipopolysaccharides; Liposomes; Lytic; MMP9 gene; Measures; Meninges; Messenger RNA; Mus; Neuroglia; Neurologic; Neutrophil Infiltration; Oral; Outcome; Pathogenicity; Pathologic; Pediatric Brain Injury; Play; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Role; Stroke; Testing; Virulence; Water; brain parenchyma; cisterna magna; cohort; cytokine; design; effective therapy; functional improvement; immunomodulatory therapies; inhibitor; insight; monocyte; motor deficit; mouse model; neonate; neutrophil; novel; post stroke; postnatal; preservation; prevent; public health relevance; single-cell RNA sequencing; tissue repair; transcriptome; transcriptome sequencing

PROJECT SUMMARY (Description) Ischemic stroke affects around 1 in 1600 to 4000 births, has the highest incidence in neonates and infants (< one year of age), and may cause neurological sequalae, including motor and cognitive deficits. Infection and immune responses are a major risk factor for pediatric stroke, but no immune-directed therapies are available. In fact, supportive measures remain the mainstay of pediatric stroke care, since the lytic treatment carries a high in neonates with fragile cerebrovascular vessels. In this project, we will use a murine model of pediatric stroke, with and without lipopolysaccharides (LPS)-sensitization, to test the pathological roles of neutrophils and monocytes towards the development of immunomodulation therapies. Aim 1: To clarify how immune cells breach the pial/glia limitans barrier in pediatric LPS/stroke. We will use flow cytometry and single-cell RNA-Seq analysis to determine the compositions and transcriptome of immune cells in the meninges after LPS/stroke, and test whether anti-MMP9 treatment prevents the neutrophil influx. Aim 2: To test whether there are two waves of monocytic infiltrates with distinct functions in pediatric stroke. We will use CCR2-CreER mice to track monocytic infiltrates and single-cell RNA-Seq analysis, as well as, block the influx of monocytes at different stage after stroke to address this issue. Aim 3: To determine what constitutes the meninges-incurred pathogenicity of neutrophils after pediatric LPS/stroke. Our RNA-Seq results suggest that neutrophils acquire IL-36gamma, a novel family of the IL-1 family cytokine, in the meninges. We will validate these findings and test the benefits of anti-IL36R treatment. Successful completion of this project will shed insights into the mechanisms of pediatric ischemic stroke and whether neutrophil/monocyte-targeted immunomodulation therapies could provide better clinical management.

项目概要（描述） 缺血性中风影响大约每 1600 至 4000 名新生儿中就有 1 人患有缺血性中风，新生儿和婴儿的发病率最高（< 一岁），并可能导致神经系统后遗症，包括运动和认知缺陷。感染和 免疫反应是小儿中风的主要危险因素，但目前尚无免疫导向疗法。 事实上，支持性措施仍然是儿科中风护理的支柱，因为溶解治疗具有一定的效果。 脑血管脆弱的新生儿发病率较高。在这个项目中，我们将使用儿科小鼠模型 中风，有或没有脂多糖（LPS）致敏，以测试中性粒细胞的病理作用 和单核细胞以开发免疫调节疗法。 目标 1：阐明儿科 LPS/中风中免疫细胞如何突破软脑膜/神经胶质限制屏障。我们将使用流 细胞计数和单细胞 RNA-Seq 分析以确定免疫细胞的组成和转录组 LPS/中风后的脑膜中，并测试抗 MMP9 治疗是否可以阻止中性粒细胞流入。 目标 2：测试小儿卒中中是否存在具有不同功能的两波单核细胞浸润。我们 将使用 CCR2-CreER 小鼠追踪单核细胞浸润和单细胞 RNA-Seq 分析，以及阻断 在中风后的不同阶段单核细胞的涌入可以解决这个问题。 目标 3：确定小儿脑膜炎后中性粒细胞致病性的构成因素 LPS/行程。我们的 RNA 测序结果表明中性粒细胞获得 IL-36gamma，这是 IL-1 的一个新家族 家族细胞因子，位于脑膜中。我们将验证这些发现并测试抗 IL36R 治疗的益处。 该项目的成功完成将深入了解小儿缺血性中风的机制和 中性粒细胞/单核细胞靶向免疫调节疗法是否可以提供更好的临床管理。