Causes and consequences of neutrophil dysfunction in early onset Crohn's disease

早发型克罗恩病中性粒细胞功能障碍的原因和后果

基本信息

批准号：
9932706
负责人：
LEE ARMISTEAD DENSON
金额：
$ 17.06万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-17 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9932706
关键词：
Abdominal colic Affect American Ancillary Study Animal Model Anti-Tumor Necrosis Factor Therapy Automobile Driving Award Biogenesis Biological Models CRISPR/Cas technology CYBA gene Cell physiology Cells Child Coculture Techniques Code Cohort Studies Colitis Complex Crohn&apos s disease DNA Methylation Data Development Dietary Supplementation Disease Enrollment Enterocolitis Epithelial Epithelial Cells Excision Exhibits Extracellular Matrix Fibrosis Functional disorder Gene Expression Gene Expression Profile Gene Mutation Genes Genetic Genetic Transcription Genetic Variation Genetic screening method Genomics Goals Health Human Hypoxia Incidence Inflammatory Inflammatory Response Intestines Knowledge Link Lipid Peroxidation Metabolic Metabolic dysfunction Methylation Microbe Missense Mutation Mitochondria Mitochondrial Proteins Modeling Molecular Molecular Profiling Mus Mutation Myofibroblast NADPH Oxidase Operative Surgical Procedures Organoids Outcome Pathway interactions Patients Pediatric Crohn&apos s disease Phenotype Physiological Play Population Precision therapeutics Process Production Property Quality of life Reactive Oxygen Species Regulation Relapse Research Role Site Small Intestines Stem cells Supplementation TGFB1 gene TNF gene Testing Untranslated RNA Variant Work Wound Healing base design early onset follow-up gastrointestinal genetic architecture genetic profiling genetic signature genetic variant genome sequencing genomic variation ileum improved induced pluripotent stem cell innovation insight loss of function mutation methylome microbial neutrophil novel novel strategies novel therapeutic intervention personalized medicine pre-clinical prebiotics preclinical study prevent programs rectal response stability testing transcriptome transcriptome sequencing whole genome

项目摘要

PROJECT SUMMARY/ABSTRACT Crohn's Disease (CD) is a relapsing and remitting gastrointestinal inflammatory disorder which can have devastating effects upon the health and quality of life of affected children. Despite the increased use of anti-TNF therapy over the past two decades, a population-level decline in rates of stricturing complications and surgeries has not been observed. The long-term goal of our multi-center research program is to define the cellular and molecular processes driving disease complications in children with CD, and to use this new knowledge to inform novel therapeutic approaches. We have made significant progress during the period of the current award in establishing a genetic basis for variation in reactive (ROS) production in pediatric CD, which we have in turn linked to stricturing complications in children. In a pre-treatment multivariable model, higher ileal expression of genes encoding mitochondrial proteins was associated with lower rates of stricturing. Studies in an animal model showed that the prebiotic 2'-Fucosyllactose would induce the protective mitochondrial gene signature. Based upon these exciting data, we hypothesize that genomic variation in NADPH oxidase ROS production regulates the cellular mitochondrial transcriptome, and 2'-Fucosyllactose primed microbial metabolites afford a novel approach to boost this protective mechanism. We propose the following complementary Aims to mechanistically test our hypotheses: • In Aim 1, we will define the ileal epithelial cell and neutrophil methylome and transcriptome in CD patients with and without NADPH oxidase complex gene mutations. These studies will define the methylome and transcriptome of ileal epithelial cells and neutrophils from CD patients carrying genetic variants associated with ROS production, and will test for stability of these signatures in patient-derived enteroids. • In Aim 2, we will test the effects of missense variants and 2'-Fucosyllactose primed microbial metabolites on ROS production and the cellular metabolic transcriptome using induced pluripotent stem cell (iPSC) derived model systems. We will differentiate iPSC carrying NADPH oxidase missense variants into neutrophils and small bowel epithelial organoids and will determine the effects of lipid peroxidation products and 2'-Fucosyllactose primed microbial metabolites upon ROS production and the mitochondrial biogenesis methylome and transcriptome in these model systems. Innovation and Impact. We will utilize primary ileal cells and novel iPSC-derived model systems to test genetic and environmental regulation of IEC and PMN ROS production and mitochondrial function, and the ability of 2'- FL primed microbial metabolites to improve cell function. These studies will provide critical insights into fundamental cellular processes regulating stricturing complications in pediatric CD, and will inform novel personalized therapies in patients classified using molecular profiles.

项目概要/摘要克罗恩病 (CD) 是一种复发性和缓解性胃肠道炎症性疾病，可导致尽管抗肿瘤坏死因子的使用有所增加，但对受影响儿童的健康和生活质量造成了毁灭性影响。过去二十年的治疗，狭窄并发症和手术率在人群水平上下降我们多中心研究计划的长期目标是定义细胞和驱动 CD 儿童疾病并发症的分子过程，并利用这一新知识来告知新颖的治疗方法在当前获奖期间取得了重大进展。为儿科 CD 中反应性 (ROS) 产生的变异建立遗传基础，我们又得到了这一基础在治疗前的多变量模型中，回肠表达较高。编码线粒体蛋白的基因与动物模型中较低的狭窄率相关。研究表明，益生元 2'-岩藻糖基乳糖会诱导保护性线粒体基因特征。根据这些令人兴奋的数据，我们探究了 NADPH 氧化酶 ROS 产生的基因组变异调节细胞线粒体转录组和 2'-岩藻糖基乳糖引发的微生物代谢物提供了一种新的我们提出以下补充目标来机械地促进这种保护机制。检验我们的假设： • 在目标 1 中，我们将定义 CD 患者的回肠上皮细胞和中性粒细胞甲基化组和转录组有或没有 NADPH 氧化酶复合物基因突变，这些研究将定义甲基化组和携带遗传变异的 CD 患者的回肠上皮细胞和中性粒细胞的转录组与 ROS 产生相关，并将测试这些特征在患者来源的肠类中的稳定性。 • 在目标 2 中，我们将测试错义变体和 2'-岩藻糖基乳糖引发的微生物代谢物的影响使用诱导多能干细胞 (iPSC) 观察 ROS 产生和细胞代谢转录组我们将携带 NADPH 氧化酶错义变体的 iPSC 分化为衍生模型系统。中性粒细胞和小肠上皮类器官，并将确定脂质过氧化的影响产品和 2'-岩藻糖基乳糖引发的微生物代谢物在 ROS 产生和线粒体这些模型系统中的生物发生甲基化组和转录组。创新和影响。我们将利用原代回肠细胞和新型 iPSC 衍生模型系统来测试遗传。 IEC 和 PMN ROS 产生和线粒体功能的环境调节，以及 2'- 的能力 FL 引发微生物代谢物以改善细胞功能。调节儿科 CD 狭窄并发症的基本细胞过程，并将为新的研究提供信息使用分子图谱分类的患者的个性化治疗。