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.

项目摘要/摘要： 语音功能障碍会损害受影响患者的生活质量并治疗这些疾病 实质性和广泛的社会，心理和经济成本。声带（VF）炎症病变 导致与上皮稳态中断有关的常见语音障碍，并伴有炎症 浸润和固有椎板的变化。在体内其他地方的上皮研究的研究中，细胞类型 起源于增生的变化是上皮干细胞。这些细胞具有自我更新的能力，并且 引起分化子细胞的后代，该细胞受局部微环境和 通过Notch信号传导细胞自主。在炎性细胞因子存在下Notch1的失活可以 导致上皮增生，可以使用体外器官进行建模。总体目标 建议是为VF上皮干细胞提供全面的特征，它们对自我的要求 生理条件下的更新和分化以及应力因素，即伤害和 机械载荷，同时还创建VF器官以阐明强调异常的分子机制 上皮重塑如良性炎症VF病变中所见。为了实现我们的目标，我们将在基因上标记 靶向LRIG1基因的上皮干细胞已与大多数上皮的干性特性有关 我们的初步数据表明，LRIG1也在人和鼠VF上皮细胞中表达。在AIM 1中， 我们将对鼠和人VF LRIG1细胞进行转录组分析，并测量LRIG1细胞响应 在稳态期间进行机械负荷。我们将描述鼠LRIG1细胞在 稳态，并在鼠LRIG1细胞中遗传灭活Notch1，以确定其对增殖的影响 体内分化。在AIM 2中，我们将在鼠模型中诱导VF上皮损伤，执行转录组 鼠LRIG1细胞的分析，并在上皮恢复期间测量其对机械负荷的反应。我们 将确定鼠LRIG1细胞和Notch1信号在重新上述化中的功能作用。在AIM 3中，我们 将使用体内下肾上腺接枝测定法确定鼠和人VF LRIG1细胞的分化潜力 和体外器官。我们将利用VF类器官对Notch介导的上皮增生进行建模 遗传，药理方法和炎症细胞因子。我们将创建一个可靠的文化系统 将提高我们对与VF上皮细胞生物学的理解 个性化医学。