ROCK, tight junctions and prematurity in the pathogenesis of necrotizing enterocolitis and neonatal sepsis.

坏死性小肠结肠炎和新生儿败血症发病机制中的 ROCK、紧密连接和早产。

• 批准号: 10659615
• 负责人: Catherine Jane Hunter
• 金额: $ 52.81万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659615
• 关键词: 12 year old; Accounting; Actins; Affect; Agonist; Apoptosis; Attenuated; Cause of Death; Cell Death; Cell physiology; Cessation of life; Clinical; Cyclic AMP-Dependent Protein Kinases; Cytoskeleton; Data; Development; Disease; E-Cadherin; Early Intervention; Endocytosis; Enterocytes; Epithelial Cells; Epithelium; Event; Excision; Experimental Models; Functional disorder; Future; Gestational Age; Health; Homeostasis; Hospitalization; Human; Immune; Impairment; Infant; Inflammation; Inflammatory; Injury; Intercellular Junctions; Intervention; Intestinal Diseases; Intestines; Knockout Mice; Knowledge; Mediating; Medical; Mission; Molecular; Mucositis; Mucous Membrane; Mus; Necrotizing Enterocolitis; Neonatal; Neonatal Intensive Care Units; Newborn Infant; Organ; Organoids; Outcome; Pathogenesis; Pathway interactions; Patient Admission; Patients; Pattern; Permeability; Predisposition; Premature Infant; Prevention strategy; Process; Protein Kinase; Protein-Serine-Threonine Kinases; Proteins; Public Health; Regulation; Research; Rho-associated kinase; Sampling; Sepsis; Signal Transduction; Specimen; Testing; Therapeutic Intervention; Tight Junctions; Tissues; United States National Institutes of Health; Work; caveolin 1; cytokine; defined contribution; feeding; genetic approach; human disease; human mortality; improved; in vivo; in vivo evaluation; insight; intestinal barrier; intestinal epithelium; microbial; microbial colonization; neonatal sepsis; neonate; occludin; pharmacologic; premature; prevent; protein expression; protein function; repaired; response; restoration; rho; systemic inflammatory response; targeted treatment; therapeutically effective; therapy development; translational potential; treatment strategy

ABSTRACT: Necrotizing enterocolitis (NEC) is a deadly disease of the newborn that affects 7% of patients admitted to the neonatal intensive care unit and is a leading cause of gram negative, neonatal sepsis. Sepsis is the second major cause of death among neonates, accounting for one million global deaths per year. Although we have made progress understanding the effect of maternal colonization patterns on neonatal sepsis, there is a gap in the understanding of the regulatory processes that regulate intestinal barrier function that contribute to susceptibility to sepsis. Indeed, the intestine is the largest immune organ in the body and is responsible for maintaining a barrier to the microbial world within its lumen. Therefore, it is a high priority for the development of medical interventions to prevent and treat neonatal sepsis. 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 found that Rho-associated protein kinase (ROCK) is activated during NEC, and inhibition of ROCK confers protection against intestinal barrier injury and disruption of normal TJ homeostasis. Our data suggest that TJ regulation, intestinal barrier function and apoptosis are regulated by a ROCK dependent, caveolin-1 mediated pathway. Furthermore, we have identified differences in TJ subcellular distribution and caveolin-1 both in patients with and without sepsis and NEC, and in those of different gestational ages. The objective of our proposed study is to define the mechanism by which barrier loss occurs in NEC and to leverage this in order to understand the potential of epithelial-specific ROCK inhibition in preventing NEC and neonatal sepsis. Our central hypothesis is that the response of the intestinal barrier in premature infants to injury is altered and is characterized by changes in epithelial ROCK signaling, aberrant TJ regulation, increased mucosal and systemic inflammation, sepsis and epithelial cell death. We will test this hypothesis by determining whether inhibition of TJ protein expression and redistribution or caveolin-1-mediated endocytosis prevent paracellular permeability increases, mucosal and systemic inflammatory responses, and epithelial cell death in experimental sepsis and NEC. The use of intestinal organoids generated from human infants and from ROCK and caveolin-1 knockout mice, will provide tremendous translational power and allow us to probe epithelial functions at molecular, cellular, tissue, and organismal levels. Using pharmacological and genetic approaches, we will determine if and how TJ responsiveness and ROCK signaling increases cell death to worsen outcomes in premature infants and whether our interventions prevent or induce sepsis. These findings have a significant positive impact on human health by providing a new understanding of the mechanisms governing epithelial intestinal barrier function of the premature infant during NEC and neonatal sepsis.

摘要： 坏死性小肠结肠炎（NEC）是一种新生儿的致命疾病，影响7%的住院患者， 新生儿重症监护病房，是革兰氏阴性菌，新生儿败血症的主要原因。脓毒症是第二个 这是新生儿死亡的主要原因，每年造成全球100万人死亡。虽然我们 在了解母体定植模式对新生儿败血症的影响方面取得了进展，但在 了解调节肠道屏障功能的调节过程， 对脓毒症的易感性。事实上，肠道是人体内最大的免疫器官， 在其内腔中保持对微生物世界的屏障。因此，发展 预防和治疗新生儿败血症的医疗干预措施。Rho激酶（ROCK）是丝氨酸/苏氨酸激酶 并参与多种细胞过程，包括调节紧密连接功能、肌动蛋白细胞骨架 收缩、炎性细胞因子和细胞死亡。我们发现Rho相关蛋白激酶（ROCK）是一种 在NEC期间激活，并且ROCK的抑制赋予针对肠屏障损伤和破坏的保护 正常的TJ内稳态我们的数据表明TJ调节，肠屏障功能和细胞凋亡是 由ROCK依赖性小窝蛋白-1介导的途径调节。此外，我们还发现了 TJ亚细胞分布和小窝蛋白-1在有和没有脓毒症和NEC的患者中，以及在那些 不同的胎龄我们提出的研究的目的是确定屏障丧失的机制， 发生在NEC，并利用这一点，以了解上皮特异性ROCK抑制的潜力， 预防NEC和新生儿败血症。我们的中心假设是，肠道屏障的反应， 早产儿对损伤的反应是改变的，其特征在于上皮ROCK信号的变化，异常TJ 调节、粘膜和全身炎症增加、败血症和上皮细胞死亡。我们将测试这个 通过确定抑制TJ蛋白表达和再分布或小窝蛋白-1介导的 内吞作用防止细胞旁通透性增加、粘膜和全身炎症反应， 实验性脓毒症和NEC中的上皮细胞死亡。使用从人类产生的肠类器官 婴儿和ROCK和caveolin-1敲除小鼠，将提供巨大的翻译能力， 在分子、细胞、组织和有机体水平上探测上皮功能。使用药理学和 通过遗传学方法，我们将确定TJ反应性和ROCK信号传导是否以及如何增加细胞死亡 早产儿的预后恶化以及我们的干预措施是否能预防或诱发败血症。这些发现 通过提供对机制的新理解，对人类健康产生重大积极影响 控制NEC和新生儿败血症期间早产儿的上皮肠屏障功能。