Single Cell Profiling and In Vivo Cellular Interrogation of Alveolar Stem Cells

肺泡干细胞的单细胞分析和体内细胞分析

基本信息

批准号：
9130386
负责人：
Tushar Jasubhai DESAI
金额：
$ 40.26万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-16 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9130386
关键词：
Ablation Acute Adult Affect Age Alveolar Alveolar Cell Alveolus Animals Bronchopulmonary Dysplasia Cancer Etiology Cell Proliferation Cell Therapy Cell physiology Cells Cessation of life Chronic Chronic Obstructive Airway Disease Clinical Development Disease Embryo Epidermal Growth Factor Epidermal Growth Factor Receptor Epithelial Epithelial Cells Etiology Exhibits Gases Genetic Genomics Goals Hamman-Rich syndrome Health Health Care Costs Heterogeneity In Vitro Individual Infant Infant respiratory distress Injury Label Life Life Cycle Stages Ligands Light Longevity Lung Lung diseases Maintenance Maps Measures Medical Molecular Morbidity - disease rate Natural regeneration Nuclear Oncogenic Pathogenesis Physiological Population Property Pulmonary Emphysema Pulmonary Fibrosis Pulmonary alveolar structure RNA Sequences Recruitment Activity Regulation Research Respiratory Failure Signal Transduction Stem cells Surface Technology Testing Therapeutic Transplantation Work adenoma alveolar epithelium attributable mortality base clinically relevant in vivo insight mortality novel progenitor prospective regenerative repaired respiratory respiratory health response self-renewal tool transcriptome sequencing treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Despite the critical importance of alveolar function for overall health and the significant morbidity and mortality attributable to diseases causing acute and chronic respiratory failure in infants and adults, little is known, or at least agreed upon, about how alveoli maintain and repair themselves throughout life. We recently used clonal analysis to map the entire life cycle of individual AT2 cells in vivo, and identified a rare subset that exhibited stem cell properties, including multi-potency, self-renewal, and activity throughout the lifespan. Here, we propose to build upon this foundational work to answer important outstanding questions about the cellular and molecular mechanisms of alveolar maintenance and stem cell regulation. Our approach employs some of the most novel and cutting-edge genetic and single cell genomic technology to accomplish the ambitious aims. Specific Aims are: A) to determine the extent and cellular mechanism of physiologic alveolar epithelial renewal in vivo by Aa) quantifying nuclear label retention by AT2 cells and Ab) measuring AT2 cell clonal fate dynamics during ageing, B) to empirically characterize molecular heterogeneity within the AT2 cell population and identify markers that distinguish AT2 stem cells Ba) using single cell RNA-sequencing of AT2 cells and Bb) by bulk cell RNA-sequencing of label-diluting versus label-retaining AT2 cells (identified in Aim Aa), C) to directly test if signaling via EGFR regulats AT2 stem cell function in vivo and whether repetitive ablation of EGFR-deleted lung epithelial cells results in alveolar enlargement or pulmonary fibrosis. By employing cutting edge technology and approaches to understand how the gas exchange surface of the lungs remain healthy throughout life, and how new cells are made to replace those that are lost from injury, we are paving the way towards developing medical or cell-based treatments to treat devastating and fatal lung diseases like emphysema and infant respiratory distress syndrome.

描述（由适用提供）：尽管肺泡功能对整体健康至关重要，并且归因于疾病的显着发病率和死亡率，导致婴儿和成人的急性和慢性呼吸衰竭，但几乎几乎是肺泡如何维持和维修肺泡如何维持和维修。我们最近使用克隆分析来绘制体内单个AT2细胞的整个生命周期，并确定了一个罕见的子集裸露的干细胞特性，包括多功能，自我更新和整个活动寿命。在这里，我们建议建立这项基础工作，以回答有关肺泡维持和干细胞调节的细胞和分子机制的重要问题。我们的方法采用了一些最新颖，最先进的遗传和单细胞基因组技术来实现雄心勃勃的目标。具体目的是：a）确定生理肺泡上皮在体内的程度和细胞机制，通过aA）通过aa）量化AT2细胞的核标记保留量，并测量AT2细胞克隆命运在衰老过程中的AT2细胞命运动力学，b）在2个细胞中的经验表征分子差异，这些分子在2个细胞中的经验表征了2个细胞的aT2细胞中的标记物，这些标记是AT2 Cell ate At2 At At aT at at at aT at aT at aT的aT2细胞的ba-ai fate a at aT 2 cell-air-ab。细胞和bb）通过散装细胞RNA的标记 - 降低与标记的at2细胞（在AIM AA中识别），c）直接测试是否通过EGFR进行信号调节Vivo中的AT2干细胞功能是否在EGFR骨中骨骼中含有肺部上皮细胞的重复性消融是否会导致肺泡增强或纯净的纯净型纤维化。通过采用尖端技术和方法来了解肺的气体交换表面如何在整个生命中保持健康，以及如何使新细胞替代因受伤而损失的细胞，我们正在壮大开发基于医学或细胞的治疗方法，以治疗毁灭性的肺部和致命性肺部疾病，例如肺气肿和婴儿呼吸窘迫综合征。