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.

项目摘要 此R03提案直接从Muir博士的研究和职业发展活动中直接得出 K08。该奖项代表了一个新的研究方向，将增强缪尔博士对 独立。该奖项将使Muir博士获得将一般改变的细胞与 三维培养系统为了定义食管上皮分化的新机制 与嗜酸性粒细胞性静脉炎（EOE）直接相关。这将在研究的指导下实现 导师，博士。 Rustgi和Nakagawa和我的K08 DRS跨学科咨询委员会。赫林（椅子）， 汤，哈克罗斯。 嗜酸性食管炎（EOE）是一种过敏性疾病，其特征是食管浸润 嗜酸性粒细胞。除了炎性细胞侵袭外，该疾病在组织学上是通过上皮的特征 变化，特别是导致基本细胞增生的上皮分化失败。在稳态下 疾病，已知食管上皮分化会受到接收器的凹陷家族的调节， 但是，在EOE的背景下，对Notch调节知之甚少。通过正在进行的K08的工作，博士 Muir研究了胶原蛋白交叉连接酶，赖赛氧化物（LOX）在EOE促进纤维化中的作用。 在本提案中提出的工作中，她现在试图确定该酶对食管的影响 上皮分化。 Muir博士的初步数据表明，LOX会影响上皮增殖 - 食道中的分化梯度。在Lox表达过表达的食管上皮细胞（EPC-LOX）中 是3D器官培养物中上皮分化的丧失，并抑制了Notch信号传导。基于这些 调查结果总体假设是LOX介导的Notch信号抑制有助于EOE 病理生物学通过限制鳞状分化和促进屏障缺陷。在强者的指导下 初步数据，该假设将通过追求两个具体目的来检验：1）确定关系 在LOX和EOE相关的上皮分化和完整性的损害之间，2）确定方式 LOX介导的Notch信号传导的调制会影响正方形分化。在第一个目标下，她会 评估LOX过表达对利用功能的增殖分化和屏障完整性的影响 细胞增殖和屏障完整性的测定。在第二个目标中，她将定义刻痕信号 在LOX过表达的背景下导致鳞状分化受损。这种方法 提案具有创新性，因为它利用了一种新型的食管3D器官培养系统，该系统概括了 食管组织学和上皮分化在体内观察到。拟议的研究很重要 试图阐明LOX在食管上皮分化中的作用，并为将来的研究提供基础 具有EOE管理中的翻译应用。