Investigating Functional Changes in the Proteins Encoded by Differentially Expressed Genes in Mouse Tracheal Epithelial Cells — 23a — Celia Brekken, Casey McKenzie, Lance Lee

Primary ciliary dyskinesia (PCD) is a rare pediatric disorder affecting one in every 16,000 children and is the result of defective motile cilia and flagella. Because of this, some common phenotypes of PCD include: respiratory issues, including sinusitis and bronchitis; hydrocephalus, accumulation of fluid in the brain; situs inversus, reversal of left-right body asymmetry; and male infertility or subfertility because of the defective sperm flagella. Unlike primary cilia, motile cilia move, beating in a wave-like motion. Each cilium consists of nine microtubule doublets surrounding a central pair of microtubules called the central pair apparatus (CPA). In our lab, we have three mouse models: nm1054 with a deletion of the gene Cfap221; Cfap54^gt/gt, a gene-trapped allele of Cfap54; bgh with a nonsense mutation in Spef2.  Previously, our lab used single-cell RNA sequencing (scRNAseq) to investigate the response of the epithelial cells in the trachea to defects in motile cilia function.  An analysis of the differentially expressed genes (DEGs) was performed and revealed important cellular information for each of the cell types. In this study, I have investigated the functional changes in the proteins that are encoded by DEGs of interest in mouse tracheal epithelial cells (mTECs) from wild type and Cfap54^gt/gt mutant mice. The cultures were differentiated, fixed, and analyzed by immunofluorescence microscopy. By comparing the subcellular localization of the proteins in the wild type mouse to the mutant mouse, we can begin to understand how these proteins and their pathways are altered in cells responding to the cilia defects.

Sanford Research
Lance Lee