Jean Francois Papon

Abnormal respiratory cilia in non-syndromic Leber congenital amaurosis with CEP290 mutations

Background Leber congenital amaurosis (LCA) is the earliest and most severe inherited retinal degeneration. Isolated forms of LCA frequently result from mutation of the CEP290 gene which is expressed in various ciliated tissues. Methods Seven LCA patients with CEP290 mutations were investigated to study otorhinolaryngologic phenotype and respiratory cilia. Nasal biopsies and brushing were performed to study cilia ultrastructure using transmission electron microscopy and ciliary beating using high-speed videomicroscopy, respectively. CEP290 expression in normal nasal epithelium was studied using real-time RT-PCR. Results When electron microscopy was feasible (5/7), high levels of respiratory cilia defects were detected. The main defects concerned dynein arms, central complex and/or peripheral microtubules. All patients had a rarefaction of ciliated cells and a variable proportion of short cilia. Frequent but moderate and heterogeneous clinical and ciliary beating abnormalities were found. CEP290 was highly expressed in the neural retina and nasal epithelial cells compared with other tissues. Discussion These data provide the first clear demonstration of respiratory cilia ultrastructural defects in LCA patients with CEP290 mutations. The frequency of these findings in LCA patients along with the high expression of CEP290 in nasal epithelium suggest that CEP290 has an important role in the proper development of both the respiratory ciliary structures and the connecting cilia of photoreceptors. The presence of respiratory symptoms in patients could represent additional clinical criteria to direct CEP290 genotyping of patients affected with the genetically heterogeneous cone-rod dystrophy subtype of LCA.

RSPH3 Mutations Cause Primary Ciliary Dyskinesia with Central-Complex Defects and a Near Absence of Radial Spokes

Primary ciliary dyskinesia (PCD) is a rare autosomal-recessive condition resulting from structural and/or functional defects of the axoneme in motile cilia and sperm flagella. The great majority of mutations identified so far involve genes whose defects result in dynein-arm anomalies. By contrast, PCD due to CC/RS defects (those in the central complex [CC] and radial spokes [RSs]), which might be difficult to diagnose, remains mostly unexplained. We identified non-ambiguous RSPH3 mutations in 5 of 48 independent families affected by CC/RS defects. RSPH3, whose ortholog in the flagellated alga Chlamydomonas reinhardtii encodes a RS-stalk protein, is mainly expressed in respiratory and testicular cells. Its protein product, which localizes within the cilia of respiratory epithelial cells, was undetectable in airway cells from an individual with RSPH3 mutations and in whom RSPH23 (a RS-neck protein) and RSPH1 and RSPH4A (RS-head proteins) were found to be still present within cilia. In the case of RSPH3 mutations, high-speed-videomicroscopy analyses revealed the coexistence of immotile cilia and motile cilia with movements of reduced amplitude. A striking feature of the ultrastructural phenotype associated with RSPH3 mutations is the near absence of detectable RSs in all cilia in combination with a variable proportion of cilia with CC defects. Overall, this study shows that RSPH3 mutations contribute to disease in more than 10% of PCD-affected individuals with CC/RS defects, thereby allowing an accurate diagnosis to be made in such cases. It also unveils the key role of RSPH3 in the proper building of RSs and the CC in humans.

Clinical characteristics, functional respiratory decline and follow-up in adult patients with primary ciliary dyskinesia

Introduction Primary ciliary dyskinesia (PCD) is a genetic disease characterised by abnormalities in ciliary function, responsible for chronic pulmonary and sinonasal diseases. Adult clinical features and outcome are poorly described. Objectives To assess the clinical characteristics and disease progression in adults with PCD. Methods Bicentric retrospective study, focusing on adults (≥18 years) with an asserted diagnosis of PCD based on the presence of bronchiectasis with typical ultrastructural defect of cilia and/or situs inversus (SI). Clinical symptoms, respiratory function, extent of bronchiectasis, microbiology and molecular analysis were assessed. Results are expressed as median (25th; 75th centile). Results 78 patients were included with a median follow-up of 8.1 years. 91% of patients had respiratory symptoms and 95% had chronic rhinosinusitis. Half of ultrastructural defects concerned dynein arms. Respiratory function was significantly lower in women (FEV1=60% predicted (50; 76), vs 77% (62; 95), p=0.009) and in patients with chronic airway Pseudomonas aeruginosa (PA, n=21) infection (FEV1=60% (48; 71) vs 75% (55; 89), p=0.04). FEV1 was associated with gender (regression coefficient for men =13.8, p=0.009), chest CT score (r=−0.42, p<0.001) but not with age at diagnosis, SI or body mass index. FEV1 decline was −13.4 mL/year (−42.8; +11.9) and was greater in women (−29.3 mL/year, (−59.7; −11.9), vs –2.0 mL/year (−26.9; +25.4), p=0.002). Three patients had severe respiratory failure. Conclusions Alteration of respiratory function in adults with PCD is heterogeneous and usually moderate but appears more severe in women and in patients with chronic PA infection. Only 4% of patients develop chronic respiratory failure.

Dossier thématiqueLa chirurgie thyroïdienne endoscopique par techniques vidéo-assistées et totalement endoscopiquesEndoscopic thyroid surgery using video-assisted and totally endoscopic techniques

R.E. Kania *, M. Parodi , A. Coste , P. Herman , P. Tran Ba Huy , J.-F. Papon b,c,d,e a CNRS, LNRS 7060, service d’ORL & CCF, pôle neurosensoriel tête et cou, hôpital Lariboisière, AP–HP, université Paris-Diderot Paris-7, 2, rue Ambroise-Paré, 75475 Paris cedex 10, France b Service ORL & CCF, hôpital Intercommunal, 94010 Créteil, France c Service ORL & CCF, groupe hospitalier Henri-Mondor – Albert-Chenevier, AP–HP, 94010 Créteil, France d Inserm, U955, 94010 Créteil, France e UMR_S841, faculté de médecine, université Paris-12, 94010 Créteil, France

La chirurgie thyroïdienne endoscopique par techniques vidéo-assistées et totalement endoscopiques

R.E. Kania *, M. Parodi , A. Coste , P. Herman , P. Tran Ba Huy , J.-F. Papon b,c,d,e a CNRS, LNRS 7060, service d’ORL & CCF, pôle neurosensoriel tête et cou, hôpital Lariboisière, AP–HP, université Paris-Diderot Paris-7, 2, rue Ambroise-Paré, 75475 Paris cedex 10, France b Service ORL & CCF, hôpital Intercommunal, 94010 Créteil, France c Service ORL & CCF, groupe hospitalier Henri-Mondor – Albert-Chenevier, AP–HP, 94010 Créteil, France d Inserm, U955, 94010 Créteil, France e UMR_S841, faculté de médecine, université Paris-12, 94010 Créteil, France

Characterization of upper airway ciliary beat by coupling isolated and collective cilia motion analysis.

Methods Ciliated cells issued from nasal brushing (controls and primary ciliary dyskinesia patients) were recorded by high-speed video-microscopy (350 frames s). We have performed an original quantitative analysis of ciliary beat dynamics (CBD) by following cilium tips. It allows to describe different parameters including ciliary beat frequency also measured by Fast-Fourier-Transform and Video-Kymography. We have also developed the microbeads tracking method (MBT) to get an index of the global efficiency of ciliary beat. Here, micro-beads (4.5 μm) have been used as markers of the flow generated by beating cilia.