Autophagy regulation of esophageal basal cell dynamics

自噬调节食管基底细胞动力学

• 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
• 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

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.

项目摘要 食管鳞状上皮包括增殖细胞的基底层，其经历分化， 基底上层和管腔脱落，促进上皮更新。虽然干细胞 为 通过我的K 01基金支持的工作， 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，自噬的作用是什么，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。我们公布的和初步的数据 在小鼠和人食管上皮的基底细胞亚群中证实了自噬的证据， 原地。此外，药理学或遗传性自噬损伤增强了3D食管类器官 离体形成。目前R 03提案的目标是研究自噬在调节细胞凋亡中的作用。 基础细胞动力学，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一个缓慢循环池， 食管干/祖细胞。为了验证这一假设，我们将追求以下具体目标：目标1： 探讨自噬水平与基底细胞干/增殖轴的关系。 食管上皮目的2：确定遗传性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的功能评估 组织活检、3D食管类器官、RNA测序和遗传工程小鼠模型， 谱系追踪能力，以研究自噬作为食管基底细胞动力学的新型调节剂， 为未来研究自噬介导的特定分子机制提供了平台。 健康和疾病条件下的食管细胞命运测定。R 03提案代表了一个逻辑 从我的K 01资助的进展寻求建立线粒体自噬作为上皮间充质细胞的关键介质 在稳态和致癌条件下的过渡，并将促进我的研究扩展 进入干细胞生物学这一激动人心的领域。这些研究将为NIH R 01提案奠定基础， 从而支持我转型为完全独立的调查员 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， 难以捉摸。