Septins in intestinal fibrosis

肠道纤维化中的脓毒症

基本信息

批准号：
10656661
负责人：
Andrei Ivanovich Ivanov
金额：
$ 63.01万
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：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10656661
关键词：
Acceleration Actomyosin Affect Animal Model Anti-Inflammatory Agents Atlases Attenuated Automobile Driving Binding Biology Cell Differentiation process Cell physiology Cells Collagen Complex Complication Crohn&apos s disease Cytoskeletal Proteins Cytoskeleton Data Deposition Development Disease Disease Progression Effector Cell Endocytosis Excision Exposure to Extracellular Matrix Extracellular Matrix Proteins Fibroblasts Fibronectins Fibrosis Filament Flagellin Fostering GTP-Binding Proteins Gene Expression Generations Genetic Genetic Induction Goals Growth Factor Health Benefit Human Immune In Vitro Incidence Inflammation Intestinal Fibrosis Intestinal Obstruction Intestines Investigation Link LoxP-flanked allele Maps Mediating Membrane Microfilaments Molecular Motor Mus Myofibroblast Myosin ATPase Operative Surgical Procedures Organ Organelles Patients Phenotype Polymers Prevention Production Property Protein Isoforms Proteins Receptor Signaling Regulation Resolution Role Signal Transduction Small Interfering RNA Structure Testing Thick Tissue Sample Tissues Transforming Growth Factors Translations United States National Institutes of Health Up-Regulation Vesicle antifibrotic treatment cell motility fibrogenesis gain of function genetic approach gut inflammation improved in vivo loss of function migration mouse model novel novel strategies novel therapeutic intervention novel therapeutics overexpression pharmacologic posttranscriptional prevent scaffold self assembly single-cell RNA sequencing therapeutically effective trafficking translatome

项目摘要

ABSTRACT Over their disease course more than half of Crohn’s disease (CD) patients develop fibrosis-induced intestinal obstruction and ultimately require surgery. No specific anti-fibrotic therapies are available. Despite advances of anti-inflammatory therapies the incidence of strictures remains high, suggesting that inflammation- independent mechanisms are crucial in the progression of the disease. The main effector cell mediating fibrosis is the myofibroblast that is activated by multiple pro-fibrotic growth factors, such as transforming growth factor (TGF)-1. Such activation results in accelerated secretion of extracellular matrix (ECM) and remodeling of the actomyosin cytoskeleton. Septins are understudied cytoskeletal proteins that regulate secretory and actomyosin- dependent cellular functions. No data on the roles and regulation of the septin cytoskeleton in intestinal fibrosis exists. Our preliminary data shows a high gene expression of septins 2, 6, 7, 8, 9, 10, 11 in human intestinal tissues with septin 7 as the most predominant isoform, which is upregulated in CD. Pharmacologic or genetic disruption of the septin cytoskeleton inhibited TGF-β1-dependent increase in ECM production (Collagen I & fibronectin) and migration in immortalized and primary human myo/fibroblasts. Preliminary evidence suggests this is post-transcriptionally regulated. Septin modulation improved experimental murine fibrosis. We hence propose to investigate the hypothesis that remodeling of the septin cytoskeleton is a driver of intestinal fibrosis and targeting the septin cytoskeleton is a novel approach to therapy of fibrostenosing Crohn’s disease. This hypothesis will be tested by three specific aims: AIM1. Characterization of alterations in septin expression and distribution in tissue samples of IBD patients. This includes development of a high-resolution map of septin expression profiles in human intestinal tissues and primary human intestinal myofibroblasts, including generation of the first full thickness single cell RNA sequencing gut atlas for stricturing CD and controls. AIM2. Investigation of the roles and mechanisms of septin dependent regulation of pro-fibrotic myofibroblast activation. We will test if septin disruption or overexpression modulates TGF-β1-signaling, intracellular vesicular trafficking or the translatome and post-transcriptionally regulated networks using a loss-of-function and gain-of- function approach. AIM3. Functional exploration if targeting septins ameliorates intestinal fibrosis in vivo. We will modulate septins in vivo using a pharmacologic and genetic approach and induce experimental fibrosis in two different animal models. We will temporally control the deletion of the central septin 7 prior to (prevention) and after induction (reversal) of experimental intestinal fibrosis specifically in Col I positive cells. If successful, this proposal will challenge the paradigm of purely immune-driven ECM deposition driving stricture formation and provide proof-of-concept for a novel mechanism to prevent or treat stricture associated intestinal obstruction in CD patients.

抽象的在病程中，超过一半的克罗恩病 (CD) 患者会出现肠道纤维化尽管取得了进展，但仍没有特定的抗纤维化疗法。在抗炎治疗中，狭窄的发生率仍然很高，这表明炎症- 独立机制在疾病进展中至关重要。主要效应细胞介导纤维化。是由多种促纤维化生长因子（例如转化生长因子）激活的肌成纤维细胞 (TGF)-1。这种激活导致细胞外基质 (ECM) 的加速分泌和细胞的重塑。肌动球蛋白细胞骨架是一种正在研究的细胞骨架蛋白，可调节分泌和肌动球蛋白。没有关于肠中隔膜细胞骨架的作用和调节的数据。我们的初步数据显示，septins 2、6、7、8、9、10、11 在人类中存在高基因表达。肠道组织中，septin 7 是最主要的异构体，在 CD 中表达上调。 septin 细胞骨架的遗传破坏抑制了 TGF-β1 依赖性 ECM 产生的增加（胶原蛋白） I 和纤连蛋白）和永生化和原代人类肌/成纤维细胞的迁移。这是转录后调节的，Septin 调节改善了实验性小鼠纤维化。提议研究以下假设：脓毒症细胞骨架的重塑是肠道的驱动因素纤维化和靶向脓毒症细胞骨架是治疗纤维狭窄克罗恩病的新方法该假设将通过三个具体目标进行检验： AIM1 的改变特征。 IBD 患者组织样本中的表达和分布这包括高分辨率的开发。人肠道组织和原代人肠道肌成纤维细胞中的 septin 表达谱图，包括生成第一个全厚度单细胞 RNA 测序肠道图谱，用于严格 CD 和对照。 AIM2. 促纤维化肌成纤维细胞的 septin 依赖性调节的作用和机制的研究我们将测试 Septin 破坏或过度表达是否会调节 TGF-β1 信号传导、细胞内囊泡。贩运或使用功能丧失和获得的翻译组和转录后调控网络 AIM3。针对败血症改善体内肠道纤维化的功能探索。使用药理学和遗传学方法在体内调节脓毒症并诱导两种实验性纤维化我们将在（预防）和之前暂时控制中央 septin 7 的删除。特别是在 Col I 阳性细胞中诱导（逆转）实验性肠纤维化后。该提案将挑战纯粹免疫驱动的 ECM 沉积驱动狭窄的范式形成并为预防或治疗相关狭窄的新机制提供概念验证 CD患者肠梗阻。