The Role of Lysyl Oxidase in Epithelial Differentiation in Eosinophilic Esophagitis

赖氨酰氧化酶在嗜酸性食管炎上皮分化中的作用

• 批准号: 9902422
• 负责人: Amanda Brooke Muir
• 金额: $ 13.2万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9902422
• 关键词: 3-Dimensional; Advisory Committees; Affect; Allergic; Allergic Disease; Automobile Driving; Award; Basal Cell Hyperplasia; Biological Assay; Biology; Cell Differentiation process; Cell Proliferation; Cells; Cellular Assay; Chronic; Collagen; Coupled; Data; Defect; Deglutition Disorders; Development; Diet; Disease; ES05; Enzymes; Eosinophilic Esophagitis; Epithelial; Epithelial Cells; Epithelium; Esophageal Squamous Cell; Esophageal Stenosis; Esophageal mucous membrane; Esophagus; Evaluation; Event; Failure; Family; Fibrosis; Flow Cytometry; Food; Functional disorder; Future; Genetic Models; Genetic Transcription; Goals; Histologic; Histology; Homeostasis; Human; Hyperplasia; Immune; Immune system; Immunoblotting; Immunosuppression; Impairment; In Vitro; Infiltration; Inflammation; Inflammatory; Interleukin-13; Intracellular Space; Lead; Luciferases; Mediating; Mentors; Methodology; Modeling; Molecular; Mus; NOTCH3 gene; Neoplasm Metastasis; Organoids; Pathology; Patients; Pattern; Pharmacology; Phenotype; Physiological; Production; Protein-Lysine 6-Oxidase; Publications; Regulation; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Squamous Differentiation; Squamous Epithelium; Steroids; Stimulus; Structure; System; TNF gene; Testing; Therapeutic; Tumor Cell Invasion; United States National Institutes of Health; Work; base; career development; crosslink; cytokine; eosinophil; feeding; gain of function; in vivo; innovation; keratinocyte; new therapeutic target; notch protein; novel; novel therapeutic intervention; overexpression; receptor; side effect; skills; standard of care; stem; tissue regeneration; transcriptome sequencing

PROJECT SUMMARY This R03 proposal stems directly from studies and career development activities outlined in Dr. Muir’s K08. This award represents a new research direction that will enhance Dr. Muir’s advancement towards independence. This award will allow Dr. Muir to obtain additional skills in combining genetically altered cells with three-dimensional culture systems in order to define novel mechanisms of esophageal epithelial differentiation with direct relevance to eosinophilic esophagitis (EoE). This will be achieved with the guidance of research mentors, Drs. Rustgi and Nakagawa and my K08 interdisciplinary advisory committee of Drs. Herlyn (Chair), Tong, and Heuckeroth. Eosinophilic esophagitis (EoE) is an allergic disease characterized by esophageal infiltration of eosinophils. In addition to inflammatory cell invasion, this disease is characterized histologically by epithelial changes, specifically failure of epithelial differentiation leading to basal cell hyperplasia. Under homeostatic conditions, esophageal epithelial differentiation is known to be regulated by the Notch family of receptors, however, little is known about Notch regulation in the context of EoE. Through the work of the ongoing K08, Dr. Muir has investigated the role of collagen cross linking enzyme, lysyl oxidase (LOX) in promoting fibrosis in EoE. In the work presented in this proposal, she now seeks to determine the effects of this enzyme on esophageal epithelial differentiation. Dr. Muir’s preliminary data demonstrate that LOX affects the epithelial proliferation- differentiation gradient in the esophagus. In LOX over-expressing esophageal epithelial cells (EPC-LOX) there is loss of epithelial differentiation in 3D organoid culture and Notch signaling is suppressed. Based on these findings the overall hypothesis is that LOX-mediated suppression of Notch signaling contributes to EoE pathobiology by limiting squamous differentiation and promoting barrier defects. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Determine the relationship between LOX and EoE-associated impairment of epithelial differentiation and integrity and 2) Determine the how LOX-mediated modulation of Notch signaling influences squamous differentiation. Under the first aim, she will evaluate the effects of LOX overexpression on proliferation-differentiation and barrier integrity utilizing functional assays of cell proliferation and barrier integrity. In the second aim, she will define how Notch signaling contributes to impaired squamous differentiation in the context of LOX over-expression. The approach in this proposal is innovative because it utilizes a novel esophageal 3D organoid culture system which recapitulates esophageal histology and epithelial differentiation observed in vivo. The proposed research is significant as it seeks to elucidate the role of LOX in esophageal epithelial differentiation and provide the basis for future studies with translational applications in the management of EoE.

项目总结 这份R03提案直接源于Muir博士的 K08。这一奖项代表了一个新的研究方向，将促进缪尔博士在 独立。这一奖项将允许缪尔博士获得更多将转基因细胞与 三维培养体系研究食道上皮细胞分化的新机制 与嗜酸性食管炎(EoE)直接相关。这将在研究的指导下实现 各位导师，Rustgi博士和Nakagawa博士以及我的K08跨学科咨询委员会Herlyn博士(主席)， 童和希克罗斯。 嗜酸性食管炎(EoE)是一种以食道浸润性食管炎为特征的过敏疾病。 嗜酸性粒细胞。除了炎性细胞的侵袭，这种疾病的组织学特征是上皮样病变。 改变，特别是上皮分化失败，导致基底细胞增生。在动态平衡下 条件下，已知的食道上皮分化受Notch受体家族调节， 然而，人们对EoE背景下的Notch监管知之甚少。通过正在进行的K08的工作，Dr。 Muir研究了胶原交联酶，赖氨酰氧化酶(LOX)在促进EoE纤维化中的作用。 在这项提案中，她现在试图确定这种酶对食道的影响 上皮分化。缪尔博士的初步数据表明，LOX影响上皮细胞的增殖- 食道的分化梯度。在LOX高表达的食道上皮细胞(EPC-LOX)中 在3D器官培养中，上皮细胞分化丧失，Notch信号被抑制。基于这些 发现总体假设是LOX介导的Notch信号抑制与EoE有关 限制鳞状细胞分化和促进屏障缺陷的病理生物学。以Strong为指导 初步数据，这一假设将通过追求两个具体目标来检验：1)确定关系 LOX和EoE相关的上皮分化和完整性损害之间的关系以及2)决定如何 LOX介导的Notch信号调节影响鳞状细胞分化。在第一个目标下，她将 应用功能评价LOX过表达对细胞增殖分化和屏障完整性的影响 细胞增殖和屏障完整性的检测。在第二个目标中，她将定义Notch信号如何 在LOX过度表达的背景下，有助于鳞状细胞分化受损。其中的方法是 该方案具有创新性，因为它使用了一种新的食道3D有机培养系统，它概括了 体内观察食道组织学和上皮分化情况。拟议的研究具有重要意义，因为它 旨在阐明LOX在食道上皮细胞分化中的作用，为今后的研究提供基础 EoE管理中的转换应用程序。