RHO-ASSOCIATED KINASE-DEPENDENT CYTOSKELETAL AND TIGHTJUNCTION DYSREGULATION IN NECROTIZING ENTEROCOLITIS

坏死性小肠结肠炎中 RHO 相关激酶依赖性细胞骨架和紧密连接失调

基本信息

批准号：
10093948
负责人：
Catherine Jane Hunter
金额：
$ 6.53万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10093948
关键词：
Actins Actomyosin Affect Animal Model Antibiotics Apoptosis Binding Cell Culture Techniques Cell Death Cell Line Cell physiology Characteristics Childhood Clinical Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytoskeleton Data Disease Emergency Situation Enterocytes Epithelial Epithelium Event Excision Functional disorder Future Goals Health Human Hypoxia In Vitro Infant Inflammation Inflammatory Intervention Intestines Knowledge Lipopolysaccharides Location Mediating Medical Mission Modeling Molecular Mucous Membrane Necrosis Necrotizing Enterocolitis Neonatal Intensive Care Units Newborn Infant Operative Surgical Procedures Outcome Oxidative Stress Pathogenesis Pathology Pathway interactions Patients Pattern Permeability Pharmacology Prevention strategy Process Production Protein Kinase Protein-Serine-Threonine Kinases Proteins Public Health Rattus Research Rest Rho-associated kinase Risk Factors Role Sampling Signal Pathway Signal Transduction Specimen Stress Structure Surgeon TLR4 gene Testing Therapeutic Therapeutic Intervention Tight Junctions Time Tissues United States National Institutes of Health Work clinically relevant cytokine feeding gastrointestinal genetic approach human disease human mortality improved improved outcome in vivo in vivo Model innovation insight intestinal barrier intestinal epithelium microbial colonization nature therapy novel novel therapeutic intervention premature preservation prevent protein distribution protein expression pup response rho targeted treatment treatment strategy

项目摘要

PROJECT SUMMARY/ ABSTRACT: Necrotizing enterocolitis (NEC) is the most common gastrointestinal emergency of newborns, and affects 7% of patients admitted to a neonatal intensive care unit. Despite years of research there is a gap in the understanding of the underlying pathophysiology of disease, and a lack of novel therapeutic approaches. Rho kinases (ROCK) are serine/ threonine kinases and are involved in multiple cellular processes including regulating tight junction function, actin cytoskeleton contraction, inflammatory cytokines and cell death. We and others, have previously demonstrated the relevance of these pathways to the pathophysiology of NEC. The objectives of this R03 proposal are to define mechanism(s) of ROCK activation, identify molecular pathways targeted by ROCK during experimental NEC and to determine the mechanisms by which ROCK inhibition limits NEC progression. The central hypothesis is that oxidative stress and LPS induce ROCK activation, resulting in cytoskeletal contraction and tight junction degradation that enhances mucosal and systemic inflammation and epithelial apoptosis. If this hypothesis is correct then ROCK inhibition will be protective against these effects and NEC. To test this hypothesis, we will examine the effects of signaling through ROCK pathway on tight junction proteins, epithelial permeability, inflammation and apoptosis during experimental NEC. The objective of this application is to define the ROCK-mediated molecular interactions that direct epithelial function during NEC. These studies will have great power since they will be performed in vitro, in enteroids, and in vivo models of NEC as well as in human intestinal samples from infants with and without NEC. We will determine whether inhibition of the ROCK pathway (by pharmacological and genetic approaches) can stabilize tight junctions and minimize inflammation, decrease cell death, and influence the outcomes and survival in experimental NEC. These findings will build upon my current studies, and have a significant positive impact on human health by providing a new understanding of the mechanisms governing epithelial intestinal barrier function during NEC.

项目摘要/摘要：坏死性小肠结肠炎 (NEC) 是新生儿最常见的胃肠道急症，影响 7% 的新生儿患者被送往新生儿重症监护病房。尽管经过多年的研究，人们的理解仍然存在差距疾病的潜在病理生理学，以及缺乏新的治疗方法。 Rho 激酶 (ROCK) 是丝氨酸/苏氨酸激酶，参与多种细胞过程，包括调节紧密连接功能、肌动蛋白细胞骨架收缩、炎症细胞因子和细胞死亡。我们和其他人之前曾证明了这些途径与 NEC 病理生理学的相关性。 R03 的目标建议定义 ROCK 激活机制，确定 ROCK 在过程中靶向的分子途径实验性 NEC 并确定 ROCK 抑制限制 NEC 进展的机制。这中心假设是氧化应激和 LPS 诱导 ROCK 激活，导致细胞骨架收缩紧密连接降解会增强粘膜和全身炎症以及上皮细胞凋亡。如果这个假设是正确的，那么 ROCK 抑制将能够防止这些影响和 NEC。为了测试这个假设，我们将检查通过 ROCK 通路的信号传导对紧密连接蛋白、上皮细胞的影响实验性 NEC 期间的通透性、炎症和细胞凋亡。该应用程序的目标是定义 ROCK 介导的分子相互作用在 NEC 期间指导上皮功能。这些研究将有强大的力量，因为它们将在 NEC 的体外、肠类和体内模型以及人类中进行来自患有和不患有 NEC 的婴儿的肠道样本。我们将确定是否抑制 ROCK 通路（通过药理学和遗传学方法）可以稳定紧密连接并最大限度地减少炎症，减少细胞死亡，并影响实验 NEC 的结果和存活。这些发现将建立在我的当前的研究，通过提供对人类健康的新认识，对人类健康产生重大积极影响 NEC 期间控制上皮肠屏障功能的机制。