Unconventional myosins and the regulation of gut barrier integrity and restitution during inflammation

非常规肌球蛋白以及炎症期间肠道屏障完整性和恢复的调节

基本信息

批准号：
10443882
负责人：
Andrei Ivanovich Ivanov
金额：
$ 45.26万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10443882
关键词：
3-Dimensional Actomyosin Acute Address Adherens Junction Adhesions Affect Air Apical Basal Cell Binding Biochemical Biological Markers Biosensor CRISPR/Cas technology Celiac Disease Cell Culture Techniques Cell Death Cell Differentiation process Cell Line Cell Polarity Cell membrane Cells Chronic Phase Colitis Complex Crohn&apos s disease Cytoskeleton Data Defect Defense Mechanisms Destinations Detergents Development Digestive System Disorders Disease Disease remission Down-Regulation Epithelial Epithelial Attachment Epithelial Cells Event Extracellular Matrix Fluorescence Resonance Energy Transfer Fractionation Gastrointestinal Diseases Genes Genetic Gut Mucosa Health Human Image Immunoprecipitation Impairment In Vitro Incidence Infectious colitis Inflammation Inflammatory Inflammatory Bowel Diseases Injury Intestinal Diseases Intestinal Mucosa Intestinal permeability Knock-out Knockout Mice Knowledge Leaky Gut Link Liquid substance Malignant Neoplasms Measurement Mediating Microfilaments Microscopy Modeling Molecular Morbidity - disease rate Morphogenesis Motor Mucositis Mucous Membrane Muscle Myosin ATPase Myosin Type II Neurons Organoids Outcome Pathogenesis Patients Periodicity Permeability Pharmacology Physiology Prevention Process Public Health Recovery Regulation Resolution Role Scaffolding Protein Signal Transduction Structure Testing Therapeutic Intervention Tight Junctions Ulcerative Colitis Western Blotting attenuation cell motility cohort cytokine disorder control epithelial injury experimental study gut homeostasis gut inflammation healing improved in vivo injured innovation insight intestinal barrier intestinal epithelium knockout animal monolayer mortality motor control mouse model mutant new therapeutic target non-muscle myosin novel overexpression pathogenic bacteria prevent repaired restoration targeted treatment tumor growth wound wound healing

项目摘要

PROJECT SUMMARY: Disruption of the intestinal epithelial barrier in patients with inflammatory bowel diseases (IBD) involves disassembly of epithelial tight junctions (TJ) and adherens junctions (AJ), along with formation of mucosal wounds due to excessive epithelial cell death. Restoration of the gut barrier (epithelial restitution) is critical for achieving remission of the disease and it involves mucosal wound healing and AJ/TJ reassembly. Both processes are mediated by common molecular events, most notably, dramatic remodeling of the epithelial actomyosin cytoskeleton. An unconventional myosin 18A (Myo18A) is a unique PDZ-domain containing scaffolding protein that acts as an essential regulator of actin filaments and their motor, non-muscle myosin II (NM II). Myo18A is known to control neuronal and muscle morphogenesis, as well as tumor growth and dissemination, however its functions in the gut have not been previously investigated. Our preliminary data suggests that Myo18A is abundantly expressed in normal human intestinal epithelium and it is a novel component of epithelial junctions, Furthermore, Myo18A depletion leads to various functional defects, including disruption of the epithelial barrier, attenuation of wound healing and impairment of 3-D epithelial morphogenesis. Importantly, we observed a marked downregulation of Myo18A expression in the intestinal epithelium of ulcerative colitis patients. This exciting preliminary data provides a strong scientific premise for the following innovative hypothesis: Myo18A is a novel regulator of the intestinal epithelial barrier integrity and restitution, and its depletion promotes gut leakiness and inhibits mucosal healing in IBD patients. This hypothesis will be tested in the following Aims: (1) to determine the roles of Myo18A in the regulation of intestinal epithelial cell migration and ECM adhesion; (2) to delineate the mechanisms of Myo18A-dependent assembly of the epithelial barrier and establishment of the apico-basal cell polarity; 3) to examine the roles of Myo18A in regulating disruption and restitution of the intestinal epithelial barrier in vivo. The Aims will be accomplished by studies utilizing in vitro intestinal epithelial cell monolayers, ex vivo colonic organoids generated from IBD mucosa and in vivo murine models of epithelial injury and restitution. Roles of Myo18A will be examined by a combination of functional (permeability measurements, wound healing), biochemical (immunoblotting, detergent fractionation), imaging (FRET biosensors, super-resolution microscopy), and genetic (CRISPR/Cas9 gene editing, overexpression of deletion mutants, knockout mice) approaches. Significance: the proposed study will yield novel insights into the mechanisms that regulate intestinal epithelial injury and restitution during inflammation. It will also identify new therapeutic targets to prevent breakdown and enhance reparation of the gut barrier in patients with digestive diseases.

项目摘要：炎症性肠患者肠上皮屏障的破坏疾病（IBD）涉及上皮紧密连接（TJ）和粘附连接（AJ）的分解，以及由于上皮细胞过度死亡而形成粘膜伤口。恢复肠道屏障（上皮细胞恢复）对于实现疾病缓解至关重要，它涉及粘膜伤口愈合和 AJ/TJ 重新组装。这两个过程都是由常见的分子事件介导的，最值得注意的是，上皮肌动球蛋白细胞骨架。非常规肌球蛋白 18A (Myo18A) 是一个独特的 PDZ 结构域含有支架蛋白，可作为肌动蛋白丝及其运动、非肌肉的重要调节剂肌球蛋白 II (NM II)。 Myo18A 已知可控制神经元和肌肉形态发生以及肿瘤生长和传播，然而其在肠道中的功能以前尚未被研究过。我们的初步数据表明Myo18A在正常人肠上皮中大量表达，并且是一种新的此外，Myo18A 耗竭会导致各种功能缺陷，包括上皮屏障破坏、伤口愈合减弱和 3-D 上皮损伤形态发生。重要的是，我们观察到肠道中 Myo18A 表达显着下调溃疡性结肠炎患者的上皮。这一令人兴奋的初步数据为以下创新假设：Myo18A 是肠上皮屏障的新型调节剂完整性和恢复性，其消耗会促进 IBD 肠道渗漏并抑制粘膜愈合患者。该假设将在以下目的中得到检验：（1）确定Myo18A在调节中的作用肠上皮细胞迁移和ECM粘附； (2) 描绘Myo18A依赖性机制上皮屏障的组装和顶端基底细胞极性的建立； 3）检查角色 Myo18A 调节体内肠上皮屏障的破坏和恢复。目标将是通过利用体外肠上皮细胞单层、离体结肠类器官的研究完成由 IBD 粘膜和上皮损伤和恢复的体内小鼠模型产生。 Myo18A 的角色将通过功能（渗透性测量、伤口愈合）、生化的组合进行检查（免疫印迹、洗涤剂分级分离）、成像（FRET 生物传感器、超分辨率显微镜）和遗传（CRISPR/Cas9 基因编辑、缺失突变体的过度表达、基因敲除小鼠）方法。意义：拟议的研究将为调节肠上皮细胞的机制提供新的见解炎症期间的损伤和恢复。它还将确定新的治疗靶点，以防止崩溃和增强消化系统疾病患者肠道屏障的修复。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Regulation of Epithelial and Endothelial Barriers by Molecular Chaperones.

分子伴侣对上皮和内皮屏障的调节。

DOI：
发表时间：
2024-02-21
期刊：
影响因子：
6
作者：
Lechuga, Susana;Marino;Naydenov, Nayden G;Zafar, Atif;Braga;Ivanov, Andrei I
通讯作者：
Ivanov, Andrei I

Unique and redundant functions of cytoplasmic actins and nonmuscle myosin II isoforms at epithelial junctions.

上皮连接处细胞质肌动蛋白和非肌肉肌球蛋白 II 亚型的独特和冗余功能。

DOI：
发表时间：
2022-09
期刊：
影响因子：
5.2
作者：
Ivanov, Andrei I;Lechuga, Susana;Marino;Naydenov, Nayden G
通讯作者：
Naydenov, Nayden G

Inflammation modulates intercellular adhesion and mechanotransduction in human epidermis via ROCK2.

炎症通过 ROCK2 调节人表皮细胞间粘附和机械转导。

DOI：
发表时间：
2023-03-17
期刊：
影响因子：
5.8
作者：
Shutova, Maria S;Borowczyk, Julia;Russo, Barbara;Sellami, Sihem;Drukala, Justyna;Wolnicki, Michal;Brembilla, Nicolo C;Kaya, Gurkan;Ivanov, Andrei I;Boehncke, Wolf
通讯作者：
Boehncke, Wolf