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Pluripotent stem cell-based modeling of cigarette smoke injury

基于多能干细胞的香烟烟雾损伤模型

基本信息

批准号：
10383130
负责人：
Kristine Abo
金额：
$ 5.1万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10383130
关键词：
3-Dimensional Acute Address Affect Air Airway Disease Apoptosis Biological Models Biology Bronchoscopy Caspase Cell Differentiation process Cells Chronic Obstructive Airway Disease Cigarette Clustered Regularly Interspaced Short Palindromic Repeats Data Data Set Diagnosis Disease Distal Epithelial Epithelial Cells Etiology Exhibits Exposure to Extrahepatic Gases Gene Expression Profile Genes Genetic Genetic Transcription Genotype HIV Seropositivity Health Human In Vitro Individual Injury Leukocyte Elastase Liquid substance Liver Liver diseases Lung Lung diseases Measures Mediating Mental Health Modeling Molecular Morbidity - disease rate Mutation NF-kappa B Organoids Pathogenesis Pathologic Pathway interactions Patients Phase Phenotype Pluripotent Stem Cells Polymers Poverty Predisposition Protein Isoforms Proteins Pulmonary Emphysema Recording of previous events Sampling Serum Proteins Signal Transduction Smoke Smoker Smoking Structure of parenchyma of lung System Testing Tissues Toxic effect United States Vulnerable Populations Work airway epithelium alpha 1-Antitrypsin alpha 1-Antitrypsin Deficiency alveolar destruction alveolar epithelium base bronchial epithelium cell type cigarette smoke differentiation protocol endoplasmic reticulum stress epithelial injury exposed human population exposure to cigarette smoke gain of function gene environment interaction in vivo induced pluripotent stem cell insight loss of function mortality non-smoker novel preventable death pulmonary function decline resilience response response to injury single-cell RNA sequencing stem cell differentiation stem cell model transcriptomics

项目摘要

Abstract Chronic obstructive pulmonary disease (COPD) is an important cause of morbidity and mortality in the US and worldwide. COPD encompasses disease of the airways as well as alveolar epithelial destruction, but the response of the alveolar epithelium to cigarette smoke has been much less well-studied than that of the airway epithelium. This is a result of poor accessibility of alveolar epithelial cells compared to airway cells, the latter of which can be obtained by bronchoscopy and brushing, and a lack of in vitro culture systems that can support the study of cigarette smoke exposure of the human alveolar epithelium. Primary human alveolar epithelial type 2 cells (AEC2s) can be cultured in three-dimensional organoids, but are difficult to culture at air-liquid interface (ALI) due to their tendency to transdifferentiate and senesce in culture. This project proposes the establishment of a novel model system to interrogate alveolar epithelial injury in response to cigarette smoke based on human induced pluripotent stem cells (iPSCs) differentiated to alveolar epithelial type 2 cells (iAEC2s) and cultured at ALI. We hypothesize that iAEC2s exhibit a distinct molecular response to cigarette smoke injury relative to syngeneic airway epithelial cells and that AATD alveolar epithelial cells are intrinsically more susceptible to cigarette smoke injury than their wild type counterparts. In aim 1, we will define the iAEC2 transcriptomic response to cigarette smoke injury. We will generate iAEC2s and expose them to gas-phase cigarette smoke at ALI. The response will be assessed by single-cell RNA sequencing of iAEC2 ALI cultures with and without cigarette smoke exposure, in comparison to the response of airway epithelial ALI cultures from the same patient. This will result in the elucidation of an alveolar epithelial-specific smoke response transcriptional signature when compared to airway epithelial cells from the same genetic background. In aim 2, we will extend our model to the only monogenic form of COPD, alpha-1 antitrypsin deficiency (AATD). We will utilize AATD-specific iPSCs that are homozygous for the polymerizing form of AAT, a genotype known as “PiZZ”, alongside their CRISPR-edited wild type isogenic counterparts (“PiMM”). We hypothesize that alpha-1 antitrypsin (AAT) is upregulated in the alveolar epithelium in response to cigarette smoke, and that PiZZ iAEC2s exhibit increased susceptibility to cigarette smoke compared to isogenic PiMM iAEC2s due to gain-of-function toxicity of the Z form of AAT that manifests as activation of the unfolded protein response and ER stress pathways. In aim 3, we will establish a PiZZ-specific transcriptomic disease signature using patient distal lung tissue. We will leverage an existing microarray dataset from PiZZ and PiMM COPD patient lung parenchymal tissue. Importantly, all patients in this study have a history of smoke exposure. We hypothesize that transcriptomic differences between the disease states will include greater pathway enrichment of the unfolded protein response and ER stress pathways in PiZZ distal lung. Completion of these aims will result in an increased understanding of lung epithelial cell type-specific responses to cigarette smoke and will add to the paradigm of disease pathogenesis in AATD.

摘要