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： 发声功能障碍损害患者的生活质量，治疗这些障碍与 巨大而广泛的社会、心理和经济代价。声带(VF)炎性病变 引起常见的嗓音障碍，与上皮细胞动态平衡紊乱伴随炎症有关 固有层渗入和改变。在对身体其他部位的上皮细胞的研究中，细胞类型 引起增生性改变的是上皮干细胞。这些细胞具有自我更新和 产生分化子细胞的后代，受局部微环境和 细胞-通过Notch信号自主地进行。在炎性细胞因子存在的情况下，Notch1的失活可能 导致上皮细胞增殖，这可以用体外有机物质来模拟。这样做的总体目标是 建议全面表征VF上皮干细胞，其对自身的要求 在生理条件下的更新和分化，以及对应激因素的响应，即损伤和 机械负荷，同时还创建VF有机化合物来阐明突出异常的分子机制 良性炎症性室性心动过速病变的上皮重塑。为了实现我们的目标，我们将在基因上贴上 靶向Lrig1基因的上皮干细胞与大多数上皮细胞的干细胞特性有关 我们的初步数据显示，Lrig1在人和小鼠的VF上皮细胞中也有表达。在目标1中， 我们将对小鼠和人类VF Lrig1细胞进行转录组谱分析，并测量Lrig1细胞的反应 动态平衡期间的机械负荷。我们将描述小鼠Lrig1细胞在 动态平衡和基因失活Notch1在小鼠Lrig1细胞中以确定其对增殖和 体内分化。在目标2中，我们将在小鼠模型中诱导室颤上皮损伤，进行转录组分析 建立小鼠Lrig1细胞的图谱，并测量其在上皮恢复过程中对机械负荷的反应。我们 将确定小鼠Lrig1细胞和Notch1信号在重新上皮化中的功能作用。在目标3中，我们 将通过体内肾下移植实验确定小鼠和人VF Lrig1细胞的分化潜能 和体外有机化合物。我们将利用VF类有机物建立Notch介导的上皮细胞增生模型 遗传学、药理学方法和炎性细胞因子。我们将建立一个可靠的文化体系， 将提高我们对VF上皮细胞生物学与VF炎性损害相关的理解 个性化医疗。