Phenotyping ciliary dynamics and coordination in response to CFTR-modulators in Cystic Fibrosis respiratory epithelial cells

Abstract

The diagnosis and treatment of respiratory disorders are challenging and would benefit from new approaches to systematically assess ciliary beating dynamics and to test new drugs. A novel approach based on multiscale differential dynamic microscopy (multi-DDM) is shown to quantitatively assess collective beating of cilia in a non-biased automated manner, in human airway epithelial cells (HAECs) derived from subjects with cystic fibrosis (CF) and grown in 2D air-liquid interface culture. Multi-DDM can readily detect changes in both ciliary beat frequency (CBF) and cilia coordination that result from perturbations to the mucosal layer. The efficacy of three CFTR-modulating treatments is investigated: ivacaftor (VX-770) with lumacaftor (VX-809), VX-809 alone and Thymosin alpha 1 (T1) alone. All three treatments restore coordination of cilia beating in the CF cells, albeit to varying degrees. We argue cilia are affected by these treatments through the physical properties of the mucus. Phenotyping cilia dynamics through multi-DDM provides novel insight into the response of ciliary beating following treatment with drugs, and has application in the broader context of respiratory disease and for drug screening.\n\nOne sentence summaryA semi-automated and unbiased assay based on multiscale differential dynamic microscopy (multi-DDM) detects changes in the coordination and frequency of ciliary beating in F508del/F508del primary human airway cells under different conditions and in response to CFTR-modulating compounds.