CRISPR/Cas9 genome editing reveals an essential role for basigin in maintaining a nonkeratinized squamous epithelium in cornea

• Published in: FASEB bioAdvances 2021-11, Vol.3 (11), p.897-908
• Main Authors: Woodward, Ashley M., Feeley, Marissa N., Rinaldi, Jamie, Argüeso, Pablo
• Format: Article
• Language: English
• Subjects: Amino acids; Antibodies; basigin; Biosynthesis; CD147 antigen; Cell culture; Cell differentiation; Cell permeability; Cloning; Cornea; COVID-19; CRISPR; CRISPR/Cas9; Electrical resistivity; Epithelial cells; Epithelium; Esophagus; Extracellular matrix; Genomes; human cornea; Matrix metalloproteinase; Metalloproteinase; nonkeratinized squamous epithelium; Plasmids; Proteins; Scanning electron microscopy; Severe acute respiratory syndrome coronavirus 2; Telomerase; Wound healing
• ISSN: 2573-9832; 2573-9832
• DOI: 10.1096/fba.2021-00067

One of the primary functions of nonkeratinized stratified squamous epithelia is to protect underlying tissues against chemical, microbial, and mechanical insult. Basigin is a transmembrane matrix metalloproteinase inducer commonly overexpressed during epithelial wound repair and cancer but whose physiological significance in normal epithelial tissue has not been fully explored. Here we used a CRISPR/Cas9 system to study the effect of basigin loss in a human cornea model of squamous epithelial differentiation. We find that epithelial cell cultures lacking basigin change shape and fail to produce a flattened squamous layer on the apical surface. This process is associated with the abnormal expression of the transcription factor SPDEF and the decreased biosynthesis of MUC16 and involucrin necessary for maintaining apical barrier function and structural integrity, respectively. Expression analysis of genes encoding tight junction proteins identified a role for basigin in promoting physiological expression of occludin and members of the claudin family. Functionally, disruption of basigin expression led to increased epithelial cell permeability as evidenced by the decrease in transepithelial electrical resistance and increase in rose bengal flux. Overall, these results suggest that basigin plays a distinct role in maintaining the normal differentiation of stratified squamous human corneal epithelium and could have potential implications to therapies targeting basigin function.