Understanding Lrig1+ in vocal fold epithelium and organoid biology

了解声带上皮和类器官生物学中的 Lrig1

• 批准号: 10732733
• 负责人: Vlasta Lungova
• 金额: $ 55.22万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10732733
• 关键词: Adult; Affect; Basal Cell; Basement membrane; Benign; Biological Assay; Biological Markers; Biology; Bromodeoxyuridine; Cell Differentiation process; Cells; Cellular biology; Data; Developmental Biology; Disease; Dyes; Ensure; Eosinophilic Esophagitis; Epithelial Cells; Epithelium; Equilibrium; Exclusion; Functional disorder; Genes; Genetic; Goals; Health; Histologic; Homeostasis; Human; Hyperplasia; Impairment; In Vitro; Inflammatory; Inflammatory Infiltrate; Injury; Label; Lamina Propria; Larynx; Lesion; Link; Maintenance; Measures; Mediating; Methods; Modeling; Molecular; Morphogenesis; Mucous Membrane; Mus; Natural regeneration; Organoids; Outcome; Patients; Physiological; Polymerase Chain Reaction; Population; Proliferating; Property; Quality of life; Recovery; Research; Role; Signal Transduction; Stratified Epithelium; Stratified Squamous Epithelium; Stratum Basale; Stress; System; Tissues; Up-Regulation; Voice; Voice Disorders; capsule; cell type; cytokine; daughter cell; economic cost; epithelial injury; epithelial stem cell; human model; improved; in vitro Model; in vivo; inflammatory milieu; insight; keratinization; mechanical load; mouse model; next generation sequencing; notch protein; personalized medicine; pharmacologic; postnatal; programs; psychologic; reconstitution; response; self-renewal; social; stem cell biology; stemness; tissue repair; transcriptomic profiling; vocal cord

Project Summary/Abstract: Voice dysfunction impairs the quality of life of affected patients and treating these disorders is associated with substantial and far-ranging social, psychological, and economic costs. Vocal fold (VF) inflammatory lesions cause common voice disorders related to disrupted epithelial homeostasis accompanied by inflammatory infiltrates and changes in the lamina propria. In studies of the epithelium from elsewhere in the body, cell types that originate hyperplasic changes are epithelial stem cells. These cells have the capability to self-renew and give rise to the progeny of differentiated daughter cells which is regulated by the local microenvironment and cell-autonomously via Notch signaling. Inactivation of Notch1 in the presence of inflammatory cytokines can lead to epithelial hyperplasia, which can be modeled using in vitro organoids. The overall objective of this proposal is to provide a comprehensive characterization of VF epithelial stem cells, their requirements for self- renewal and differentiation under physiological conditions and in response to stress factors, namely injury and mechanical load, while also creating VF organoids to elucidate molecular mechanisms that underline aberrant epithelial remodeling as seen in benign inflammatory VF lesions. To achieve our goal we will genetically label epithelial stem cells targeting the Lrig1 gene that has been linked to stemness properties in majority of epithelia and our preliminary data show that Lrig1 is also expressed in human and murine VF epithelial cells. In Aim 1, we will perform transcriptome profiling of murine and human VF Lrig1 cells and measure Lrig1 cell responses to mechanical load during homeostasis. We will delineate the mechanistic role of murine Lrig1 cells in homeostasis, and genetically inactivate Notch1 in murine Lrig1 cells to determine its effect on proliferation and differentiation in vivo. In Aim 2, we will induce VF epithelial injury in a murine model, perform transcriptome profiling of murine Lrig1 cells and measure their responses to mechanical load during epithelial recovery. We will determine the functional role of murine Lrig1 cells and Notch1 signaling in re-epithelization. In Aim 3, we will determine differentiation potential of murine and human VF Lrig1 cells using in vivo subrenal graft assay and in vitro organoids. We will utilize VF organoids to model Notch-mediated epithelial hyperplasia using genetic, pharmacologic approaches, and inflammatory cytokines. We will create a reliable culture system that will improve our understanding of VF epithelial cell biology related to VF inflammatory lesions in the context of personalized medicine.

项目概要/摘要： 声音功能障碍损害受影响患者的生活质量，治疗这些疾病与 社会、心理和经济代价巨大而深远。声带（VF）炎性病变 引起与上皮稳态破坏相关的常见声音障碍，伴有炎症 浸润和固有层的变化。在对身体其他部位上皮细胞的研究中， 是上皮干细胞。这些细胞具有自我更新的能力， 产生受局部微环境调节的分化子细胞的后代， 通过Notch信令进行小区自主。在炎症细胞因子存在下Notch 1的失活可以 导致上皮增生，这可以使用体外类器官来建模。本报告的总体目标 建议是提供VF上皮干细胞的全面表征，它们对自身免疫的要求， 在生理条件下和响应于应激因素，即损伤和 机械负荷，同时也创造VF类器官，以阐明强调异常的分子机制， 良性炎性VF病变中所见的上皮重塑。为了实现我们的目标，我们将在基因上标记 靶向Lrig 1基因的上皮干细胞，该基因与大多数上皮细胞的干性特性有关 我们的初步数据显示Lrig 1也在人和鼠VF上皮细胞中表达。在目标1中， 我们将对鼠和人VF Lrig 1细胞进行转录组分析，并测量Lrig 1细胞反应， 承受机械负荷的能力我们将描述小鼠Lrig 1细胞在 稳态，并在鼠Lrig 1细胞中遗传标记Notch 1，以确定其对增殖和 体内分化。在目标2中，我们将在小鼠模型中诱导VF上皮损伤，进行转录组学研究， 分析鼠Lrig 1细胞并测量它们在上皮恢复期间对机械负荷的响应。我们 将确定鼠Lrig 1细胞和Notch 1信号传导在再上皮化中的功能作用。在目标3中，我们 将使用体内肾下移植物测定法测定鼠和人VF Lrig 1细胞的分化潜能 和体外类器官。我们将利用VF类器官来模拟Notch介导的上皮增生， 遗传学、药理学方法和炎症细胞因子。我们将建立一个可靠的文化体系， 将提高我们对VF上皮细胞生物学的理解， 个性化医疗