Gain-of-function Variants in SMAD4 compromise Respiratory Epithelial cells

To investigate how SMAD4 mutations associated with Myhre syndrome impact SMAD4 protein levels, activation and physiological functions in patient derived nasal epithelial cells. We derived primary nasal basal stem cells from inferior turbinate brushings of healthy subjects and Myhre syndrome patients (SMAD4-Ile500Val, Arg496Cys, Ile500Thr). After ALI differentiation, RNA was collected from healthy nasal basal epithelial cells and Myhre syndrome-epithelial cells to perform mRNA Deep sequencing. We then performed gene expression profiling analysis using data obtained from RNA-seq. Total RNA was extracted from nasal basal epithelial cells (passage P1). Comparative gene expression profiling analysis was done of RNA-seq data for nasal basal epithelial cells and Myhre syndrome-epithelial cells.本研究旨在探究与Myhre综合征相关的SMAD4突变，在患者来源的鼻上皮细胞中对SMAD4蛋白水平、激活状态及生理功能的影响机制。我们从健康受试者与Myhre综合征患者（携带SMAD4-Ile500Val、Arg496Cys、Ile500Thr突变）的下鼻甲刷取样本中分离原代鼻基底干细胞。经气液界面（Air-Liquid Interface, ALI）分化培养后，分别收集健康鼻基底上皮细胞与Myhre综合征源性上皮细胞的RNA，开展mRNA深度测序。随后基于RNA测序（RNA-seq）获得的数据进行基因表达谱分析。从第1代（P1）鼻基底上皮细胞中提取总RNA。针对健康鼻基底上皮细胞与Myhre综合征源性上皮细胞的RNA-seq数据，完成对比性基因表达谱分析。