Dissecting the Functional Role of ASCL1 in the Human Thymus and Airway in Vitro

Abstract

Abstract This study investigates the role of ASCL1 gene deletion in differentiating Human Thymic Epithelial Cells (HTECs) and Human Bronchial Epithelial Cells (HBECs). It aims to understand ASCL1's impact on cellular differentiation and its therapeutic implications. Flow cytometry evaluated transfection efficiency, indicating higher efficacy in HTECs than in HBECs. ASCL1 knockout validation was achieved through Western blot analysis. Gene expression analyses using RT-qPCR unveiled distinct downregulation patterns in HTECs and HBECs upon ASCL1 deletion. The study highlights ASCL1's significance in regulating genes crucial for cortical and medullary differentiation in HTECs including FOXN1, CSTV, FOXI1, SOX2, SYP and CLDN4 whereas the expression of LY75 remained unchanged. Also, the expression of SYP in HBECs showed a significant reduction although we did not notice the deletion of ASCL1. These findings expand our understanding of ASCL1's multifaceted role in guiding epithelial cell differentiation, offering potential prospects for regenerative medicine and targeted therapies for epithelial tissue-related disorders.