Evaluation of Functional Activity of Ciliated Epithelium in Patients with Primary Ciliary Dyskinesia
Tatiana Kyian, Anna Demchenko, Olga Solovova, Maxim Tarasov, Svetlana Smirnikhina, Aleksandra Nazarova, Elizaveta Bragina, Oksana Ryzhkova, Elena Kondratyeva, Sergey KutsevPrimary ciliary dyskinesia (PCD) is a heterogeneous recessive disorder of cilia that leads to respiratory tract disorders and organ laterality defects. Objectives: This study was designed to diagnose PCD in patients using various methods: genetic, immunofluorescence, transmission electron microscopy (TEM), and assessment of ciliary beat frequency (CBF) in ALI cultures. Methods: The study included (n = 35) patients with primary ciliary dyskinesia and (n = 10) healthy controls (HC). The diagnosis of PCD was established using molecular genetic analysis: WES, TEM, immunofluorescence staining of ALI-culture, and high-speed videomicroscopy. Ciliogenesis of ciliated epithelial cells was performed in patients with PCD and HC to improve the diagnosis of PCD. Results: Genetic sequencing was conducted for all patients and variants in 17 genes were found. All variants were known. A comparison of CBF assessment in PCD patients and HC using high-speed videomicroscopy showed that the CBF in PCD patient nasal cells was significantly reduced (p < 0.0001), with sensitivity of 91% and specificity of 89.2%. CBF was independent of age and showed no statistical difference between adults and children. Conclusions: The diagnosis of PCD was confirmed in all patients using various diagnostic methods, including the assessment of CBF of ALI culture. The creation of databases of ALI cultures, combined with genotypic and structural abnormalities, is necessary both for diagnosis and, in the future, for use as models in the development of targeted therapies and for assessing their effectiveness.