Autophagy regulation of esophageal basal cell dynamics

自噬调节食管基底细胞动力学

基本信息

批准号：
9586311
负责人：
Kelly A Whelan
金额：
$ 11.89万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9586311
关键词：
Area Autophagocytosis Basal Cell Biological Assay Biology Biomedical Research Biopsy CRISPR/Cas technology Cell Compartmentation Cell Culture Techniques Cell Cycle Cell Fraction Cell Lineage Cell Proliferation Cells Cellular biology Coupling Data Diagnosis Disease Dyes Endoscopic Biopsy Epithelial Epithelium Equilibrium Esophageal Esophageal Diseases Esophagitis Esophagus Evaluation Fluorescence-Activated Cell Sorting Foundations Future Genetic Genetic Transcription Genetically Engineered Mouse Goals Grant Growth Health Homeostasis Human Impairment In Situ Investigation Kinetics Knowledge Maintenance Malignant Neoplasms Maps Mediating Mediator of activation protein Mesenchymal Methods Mitochondria Molecular Monitor Mus Organoids Oxidative Stress Patients Pharmacology Play Population Proliferating Publishing Regulation Research Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Specimen Squamous Epithelium Stains Stem cells Stratum Basale Tamoxifen Testing Tissues United States National Institutes of Health Vesicle Work cancer stem cell carcinogenesis collaborative approach improved innovation keratinocyte novel progenitor programs self-renewal stem stem cell biology stem cell population stemness terminally differentiated effector memory (TEM) T cells transcriptome transcriptome sequencing tumor microenvironment

项目摘要

Project Summary Esophageal squamous epithelium comprises a basal layer of proliferative cells that undergoes differentiation in the suprabasal layer and luminal desquamation, facilitating epithelial renewal. Although stem cells responsible for and Through work supported by my K01 grant, we have recently demonstrated that autophagy-mediated modulation of oxidative stress and mitochondrial function supports expansion of cancer stem cells in the tumor microenvironment; however, what role, if any, autophagy plays in normal esophageal stem cell biology remains to be determined. Our published and preliminary data demonstrate evidence of autophagy in a subset of basal cells in murine and human esophageal epithelium in situ. Additionally, pharmacologic or genetic autophagy impairment enhances 3D esophageal organoid formation ex vivo. The goal of the current R03 proposal is to investigate the role of autophagy in regulating basal cell dynamics, namely the balance between actively proliferating and quiescent stem cells. We hypothesize that autophagy limits basal cell proliferation to maintain a pool of slow-cycling esophageal stem/progenitor cells. To test this hypothesis we will pursue the following Specific Aims: Aim 1: To determine the relation relationship between autophagy level and the basal cell stemness/proliferation axis in esophageal epithelium. Aim 2: To define the functional consequences of genetic autophagy impairment upon esophageal basal cell dynamics. These innovative studies utilize functional evaluation of human endoscopic tissue biopsies, 3D esophageal organoids, RNA-sequencing, and a genetically engineered murine model with lineage tracing capacity to investigate autophagy as a novel regulator of esophageal basal cell dynamics, building a platform for future investigations into the specific molecular mechanisms underlying autophagy-mediated esophageal cell fate determination under conditions of health and disease. This R03 proposal represents a logical progression from my K01 grant seeking to establish mitophagy as a critical mediator of epithelial-mesenchymal transition under conditions of homeostasis and carcinogenesis and will facilitate expansion of my research program into the exciting field of stem cell biology. These studies will lay the foundation for an NIH R01 proposal, thereby supporting my transition to a fully independent investigator. the maintenance of esophageal epithelium characterization of these cells remains are thought to reside in the basal cell compartment, identification elusive.

项目概要食管鳞状上皮包含增殖细胞的基底层，这些细胞在基底上层和腔内剥落，促进上皮更新。尽管干细胞负责为了通过我的 K01 补助金支持的工作，我们已经最近证明自噬介导的氧化应激和线粒体功能的调节支持肿瘤微环境中癌症干细胞的扩增；然而，自噬有何作用（如果有的话）在正常食管干细胞生物学中的作用仍有待确定。我们公布的初步数据证明小鼠和人类食管上皮的基底细胞亚群存在自噬的证据现场。此外，药理学或遗传自噬损伤可增强 3D 食管类器官离体形成。当前 R03 提案的目标是研究自噬在调节中的作用基底细胞动力学，即活跃增殖和静止干细胞之间的平衡。我们假设自噬限制基底细胞增殖以维持缓慢循环的池食管干/祖细胞。为了检验这一假设，我们将追求以下具体目标：目标 1：确定自噬水平与基底细胞干性/增殖轴之间的关系食管上皮。目标 2：定义遗传自噬损伤的功能后果食管基底细胞动力学。这些创新研究利用人体内窥镜功能评估组织活检、3D 食管类器官、RNA 测序和基因工程小鼠模型谱系追踪能力研究自噬作为食管基底细胞动力学的新型调节剂，建立未来研究自噬介导的特定分子机制的平台健康和疾病条件下食管细胞的命运决定。这个 R03 提案代表了一个合乎逻辑的我的 K01 资助的进展，旨在将线粒体自噬确立为上皮间充质的关键介质稳态和致癌条件下的转变，将有助于扩大我的研究计划进入令人兴奋的干细胞生物学领域。这些研究将为 NIH R01 提案奠定基础，从而支持我向完全独立的调查员过渡。食管上皮的维护这些细胞的特征仍然存在被认为存在于基底细胞室中，鉴定难以捉摸。