An Integrated Analysis of Radial Spoke Head and Outer Dynein Arm Protein Defects and Ciliogenesis Abnormality in Nasal Polyps

Abstract

Background: Nasal polyp (NP) is a chronic upper airway inflammatory disease that is frequently triggered by defective host-defense. However, the mechanisms underlying the impaired barrier function such as cilia-mediated mucociliary clearance remain poorly understood. Objective: To assess ciliary ultrastructural and ciliogenesis marker expression and the phenotypes of ciliated cells in NP. Methods: NP biopsy samples were obtained from 97 NP patients and inferior turbinate from 32 healthy controls. Immunofluorescence staining, quantitative polymerase chain reaction, and single-cell cytospin staining were performed. We classified the patterns of radial spoke head protein (RSPH) 1, 4A (RSPH4A), 9 (RSPH9), and dynein axonemal heavy chain 5 (DNAH5) localization. A semi-quantitative scoring system was developed to assess their expression patterns and associations with ciliogenesis markers [centrosomal protein 110 (CP110) and forkhead box j1 (FOXJ1)]. Results: Median scores of RSPH1, RSPH4A, RSPH9, and DNAH5 were significantly higher in NP than in healthy controls, particularly in eosinophilic NPs. Expression pattern scores of RSPH1, RSPH4A, RSPH9, and DNAH5 correlated positively with each other in both groups. In primary-cell specimens, abnormal expression patterns were significantly more common in NP. The total fluorescence intensity of CP110 and FOXJ1 was significantly higher in NPs and correlated positively with expression pattern scores of RSPH1, RSPH4A, RSPH9, and DNAH5. A trend towards lengthened cilia was observed in NP. Conclusion: In the chronic airway inflammatory milieu, the up-regulated ciliogenesis correlates with the abnormal expression of ciliary ultrastructural markers (i.e., DNAH5) in NP (particularly eosinophilic NP).

An Integrated Analysis Reveals Ciliary Abnormalities in Antrochoanal Polyps

“…Abnormal expression of ciliary ultrastructural markers has been implicated in chronic lower airway inflammatory diseases that have previously been linked to genetic defects, such as DNAH5, DNALI1, GAS8 (a component of the nexin-dynein regulatory complex), RSPH4A, RSPH9 and RSPH1 . 24 , 25 According to literatures, secondary ciliary dyskinesia was found in patients with chronic rhinosinusitis, 7 , 8 , 26 , 27 bronchiectasis 28 and chronic obstructive pulmonary disease. 29 The abnormal ciliary ultrastructure consisted mainly of compound cilia, microtubular and dynein arm defects.…”

“… 27 Our published study has demonstrated the abnormal-regulation of ciliogenesis markers ( CP110 and FOXJ1 ) and overly dense and lengthened cilia, greater prevalence of absence of DNAH5, RSPH1, RSPH4A and RSPH9 expressions, and the significantly reduced ciliary beat frequency have also been observed in NPs in the aberrantly remodeled epithelium of NPs. 7 , 8 , 26 However, the characteristics of ciliary impairment in ACPs remain not fully explored. Therefore, according to the result of our sequencing analysis, we evaluated the expression of 9 common cilia-related genes and 2 ciliogenesis markers by qPCR analysis and IF staining.…”

“…It has been reported that MCC abnormality of the airway epithelium has been closely associated with the pathogenesis of several airway inflammatory disease (eg, allergic rhinitis, nasal polyps, asthma and idiopathic pulmonary fibrosis). [7][8][9][10][11] However, it remains elusive whether the MCC abnormalities are common pathologic characteristics in ACP patients, and further studies to detect the expression levels of MCC related markers as well as to characterize the morphological features of ciliated cells in ACP tissues are warranted.…”

“…Besides, the TFI of MUC5AC and KRT5 expression was non-significantly greater in ACPs than in control [5.05×10 6 (2.52×10 6 , 8.31×10 6 ) vs 3,391,055 (1.03×10 6 , 1.23×10 7 ), P = 0.443; 2.51×10 7 (2.22×10 7 , 3.17×10 7 ) vs 2.20×10 7 (1.59×10 7 , 2.68×10 7 ), P = 0.208] (Figure 4).…”

Ciliary Function, Antigen Stasis and Asthma

“…Note also that abnormal expression patterns of RSPH1, RSPH4A, RSPH9, and DNAH5 were significantly more common in patients with nasal polyps, a chronic upper airway inflammatory disease that is frequently triggered by defective host-defense [58], even in the absence of classic PCD.…”