Modulation of choroid plexus immuno-secretory function to restore cerebrospinal fluid homeostasis in hydrocephalus

调节脉络丛免疫分泌功能以恢复脑积水的脑脊液稳态

• 批准号: 10247073
• 负责人: Kristopher Kahle
• 金额: $ 36.64万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247073
• 关键词: Acute; Affect; Animals; Antisense Oligonucleotides; Attenuated; Bacterial Meningitis; Binding; Biochemical; Biological Assay; Blood; Brain; Brain imaging; Carrier Proteins; Cell Wall; Cells; Cerebral Ventricles; Cerebrospinal Fluid; Cerebrospinal fluid shunts procedure; Choroid Plexus Epithelium; Clinical Trials Design; Co-Immunoprecipitations; Complication; Data; Dependence; Development; Disease; Drug Delivery Systems; Drug Formulations; Epithelial; Failure; Flagellin; Fluid Balance; Fluids and Secretions; Functional disorder; Future; Goals; Healthcare Systems; Hemorrhage; Hydrocephalus; Immune; Immunohistochemistry; In Vitro; Individual; Infection prevention; Inflammation; Inflammatory; Ion Transport; Ions; Knock-out; Knowledge; Lipopolysaccharides; Live Birth; Magnetic Resonance Imaging; Measurement; Mediating; Mediator of activation protein; Medicine; Methemoglobin; Methodology; Modeling; Molecular; Morbidity - disease rate; Nature; Operative Surgical Procedures; Pathogenesis; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Pharmacology; Phospho-Specific Antibodies; Phosphorylation; Phosphotransferases; Physiological; Physiology; Play; Plumbing; Prevention; Proteome; Publishing; Rattus; Role; Shunt Device; Signal Transduction; Structure of choroid plexus; TLR1 gene; TLR4 gene; TLR5 gene; Techniques; Testing; Therapeutic; Treatment Efficacy; United States National Institutes of Health; Ventricular; Work; base; cost; cytokine; improved; in vivo; inhibitor/antagonist; innovation; insight; intraventricular hemorrhage; mortality; neuroinflammation; novel; novel therapeutic intervention; pathogen; phosphoproteomics; preclinical study; prevent; public health relevance; real time monitoring; response; symposium; targeted treatment; therapeutic evaluation

PROJECT SUMMARY A recent NIH-sponsored Hydrocephalus Symposium highlighted the critical unmet need to develop effective non-surgical hydrocephalus therapies based on improved understanding of the choroid plexus epithelium (CPe) and its mechanisms of cerebrospinal fluid (CSF) secretion. These knowledge gaps perpetuate current reliance CSF shunting surgeries with high morbidity and failure rates. The scientific premise of this proposal is based on our recently published (Nature Medicine, 2017) and unpublished data suggesting the CPe’s immuno- secretory plasticity plays an essential role in the pathogenesis of acute post-hemorrhagic hydrocephalus (PHH) via a toll-like receptor-4 (TLR4)-dependent increase in CSF secretion regulated by the NF-κB-regulated SPAK kinase. However, several fundamental questions require elucidation: (i) How does intraventricular hemorrhage (IVH) cause CPe inflammation? (ii) Which CPe ion transporters are required for the CSF hypersecretory response? (iii) Does this mechanism contribute to the pathogenesis of post-infectious hydrocephalus (PIH)? (iv) Can drug inhibition of TLR4, SPAK, or other CPe targets post-IVH or infection prevent or attenuate hydrocephalus? These questions frame our central hypothesis that CSF-borne damage- and pathogen- associated molecular patterns (DAMPs and PAMPs) associated with IVH (methemoglobin) and bacterial meningitis (lipopolysaccharide [LPS]), stimulate TLR4/MyD88 signaling to cause CPe inflammation, and the associated CSF hypersecretory response requires a functional ensemble of SPAK-regulated CPe ion transport proteins. This hypothesis will be tested in 3 specific aims: (1) elucidate the TLR4-dependent CPe inflammatory mechanism(s) triggered by IVH; (2) identify the TLR4-dependent CPe ion transport effectors that mediate IVH- induced CSF hypersecretion; and (3) characterize the effects of bacterial PAMPs central to PIH on CPe immuno-secretory function. To do this, we will use wild type and TLR4 knockout rats in a validated model of PHH, and our novel LPS-induced model of PIH, and employ direct in vivo real-time monitoring of CSF secretion; non-invasive MR imaging of ventricular volume in live animals; quantitative CPe phospho- proteomics to interrogate signaling networks; and the intracerebroventricular delivery of drugs and antisense oligonucleotides to modulate CPe targets. Our study's overall objective is to identify specific CPe inflammatory and/or ion transport proteins that can be pharmacologically leveraged to prevent hydrocephalus, thereby bringing us nearer to our long-term goal of eliminating surgical shunt dependence. Our proposal is innovative because it challenges the status quo conceptual, methodological, and therapeutic approaches to hydrocephalus. If successful, this work could catalyze a change in our view of hydrocephalus from a neurosurgical “brain plumbing” disorder to a drug-preventable neuro-inflammatory condition. In advancing our basic understanding of CPe immuno-secretory function, this work may also inform development of novel therapeutic strategies for other conditions associated with neuroinflammation or disordered CSF dynamics.

项目概要 最近由美国国立卫生研究院主办的脑积水研讨会强调了开发有效的脑积水的关键未满足的需求 基于对脉络丛上皮的了解的非手术脑积水治疗 （CPe）及其脑脊液（CSF）分泌机制。这些知识差距使当前的 依赖脑脊液分流手术的发病率和失败率很高。该提案的科学前提是 基于我们最近发表的（Nature Medicine，2017）和未发表的数据表明 CPe 的免疫 分泌可塑性在急性出血后脑积水（PHH）的发病机制中起着重要作用 通过由 NF-κB 调节的 SPAK 调节的 Toll 样受体 4 (TLR4) 依赖性脑脊液分泌增加 激酶。然而，有几个基本问​​题需要阐明：（i）脑室内出血是如何发生的？ （IVH）引起CPe炎症？ (ii) CSF 过度分泌需要哪些 CPe 离子转运蛋白 回复？ (iii) 这种机制是否有助于感染后脑积水（PIH）的发病机制？ (iv) 药物抑制 TLR4、SPAK 或其他 CPe 靶点是否可以预防或减弱 IVH 后或感染的效果 脑积水？这些问题构成了我们的中心假设，即脑脊液引起的损伤和病原体 与 IVH（高铁血红蛋白）和细菌相关的相关分子模式（DAMP 和 PAMP） 脑膜炎（脂多糖 [LPS]），刺激 TLR4/MyD88 信号传导导致 CPe 炎症，以及 相关的 CSF 分泌过多反应需要 SPAK 调节的 CPe 离子转运的功能整体 蛋白质。该假设将在 3 个具体目标中得到检验：(1) 阐明 TLR4 依赖性 CPe 炎症 IVH 触发的机制； (2) 鉴定介导 IVH- 的 TLR4 依赖性 CPe 离子转运效应子 诱导脑脊液分泌过多； (3) 表征对 PIH 至关重要的细菌 PAMP 对 CPe 的影响 免疫分泌功能。为此，我们将在经过验证的模型中使用野生型和 TLR4 敲除大鼠 PHH 和我们的新型 LPS 诱导的 PIH 模型，并采用直接体内实时监测 CSF 分泌;活体动物心室容积的无创 MR 成像；定量 CPe 磷酸化 蛋白质组学研究信号网络；以及药物和反义药物的脑室内递送 寡核苷酸来调节 CPe 靶标。我们研究的总体目标是确定特定的 CPe 炎症 和/或离子转运蛋白，可以在药理学上利用来预防脑积水，从而 使我们更接近消除手术分流依赖的长期目标。我们的建议很创新 因为它挑战了现状的概念、方法和治疗方法 脑积水。如果成功，这项工作可能会促使我们对脑积水的看法发生改变 神经外科“大脑管道”紊乱导致药物可预防的神经炎症性疾病。在推进我们的 对 CPe 免疫分泌功能的基本了解，这项工作也可能为新型药物的开发提供信息 与神经炎症或脑脊液动力学紊乱相关的其他疾病的治疗策略。