title: Immunofluorescence Analysis and Diagnosis of Primary Ciliary Dyskinesia with Radial Spoke Defects.
abstract: Primary ciliary dyskinesia (PCD) is a genetically heterogeneous recessive disorder caused by several distinct defects in genes responsible for ciliary beating, leading to defective mucociliary clearance often associated with randomization of left/right body asymmetry. Individuals with PCD caused by defective radial spoke (RS) heads are difficult to diagnose owing to lack of gross ultrastructural defects and absence of situs inversus. Thus far, most mutations identified in human radial spoke genes (RSPH) are loss-of-function mutations, and missense variants have been rarely described. We studied the consequences of different RSPH9, RSPH4A, and RSPH1 mutations on the assembly of the RS complex to improve diagnostics in PCD. We report 21 individuals with PCD (16 families) with biallelic mutations in RSPH9, RSPH4A, and RSPH1, including seven novel mutations comprising missense variants, and performed high-resolution immunofluorescence analysis of human respiratory cilia. Missense variants are frequent genetic defects in PCD with RS defects. Absence of RSPH4A due to mutations in RSPH4A results in deficient axonemal assembly of the RS head components RSPH1 and RSPH9. RSPH1 mutant cilia, lacking RSPH1, fail to assemble RSPH9, whereas RSPH9 mutations result in axonemal absence of RSPH9, but do not affect the assembly of the other head proteins, RSPH1 and RSPH4A. Interestingly, our results were identical in individuals carrying loss-of-function mutations, missense variants, or one amino acid deletion. Immunofluorescence analysis can improve diagnosis of PCD in patients with loss-of-function mutations as well as missense variants. RSPH4A is the core protein of the RS head.
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Adrien
authors: A
authors: Frommer
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Rim
authors: R
authors: Hjeij
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Niki T
authors: NT
authors: Loges
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Christine
authors: C
authors: Edelbusch
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Charlotte
authors: C
authors: Jahnke
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Johanna
authors: J
authors: Raidt
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Claudius
authors: C
authors: Werner
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Julia
authors: J
authors: Wallmeier
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Jörg
authors: J
authors: Große-Onnebrink
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Heike
authors: H
authors: Olbrich
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Sandra
authors: S
authors: Cindrić
authors: 2 Department of Otorhinolaryngology, University Hospital Leuven, Leuven, Belgium.
authors: Martine
authors: M
authors: Jaspers
authors: 3 Department of Pediatrics, Pediatric Pulmonology, University Hospital of Leuven, Leuven, Belgium.
authors: Mieke
authors: M
authors: Boon
authors: 4 Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom.
authors: Yasin
authors: Y
authors: Memari
authors: 4 Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom.
authors: Richard
authors: R
authors: Durbin
authors: 4 Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom.
authors: Anja
authors: A
authors: Kolb-Kokocinski
authors: 5 Max-Planck-Institute for Molecular Genetics, Berlin, Germany.
authors: Sascha
authors: S
authors: Sauer
authors: 6 Danish Primary Ciliary Dyskinesia (PCD) Centre and Pediatrics Pulmonary Service, Department of Pediatrics and Adolescent Medicine, Copenhagen University Hospital, Rigshospitalet, Denmark.
authors: June K
authors: JK
authors: Marthin
authors: 6 Danish Primary Ciliary Dyskinesia (PCD) Centre and Pediatrics Pulmonary Service, Department of Pediatrics and Adolescent Medicine, Copenhagen University Hospital, Rigshospitalet, Denmark.
authors: Kim G
authors: KG
authors: Nielsen
authors: 7 Department of Pediatrics, University of Alberta, Edmonton, Alberta, Canada.
authors: Israel
authors: I
authors: Amirav
authors: 8 Saint Vincent De-Paul Hospital, Nazareth, Israel.
authors: Nael
authors: N
authors: Elias
authors: 9 Cystic Fibrosis and PCD Center, Hadassah Hebrew University Hospital, Jerusalem, Israel; and.
authors: Eitan
authors: E
authors: Kerem
authors: 9 Cystic Fibrosis and PCD Center, Hadassah Hebrew University Hospital, Jerusalem, Israel; and.
authors: David
authors: D
authors: Shoseyov
authors: 10 Department of Pediatrics, University Hospital Freiburg, Freiburg, Germany.
authors: Karsten
authors: K
authors: Haeffner
authors: 1 Department of General Pediatrics, University Children's Hospital Muenster, Muenster, Germany.
authors: Heymut
authors: H
authors: Omran
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007619:Kartagener Syndrome / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D008297:Male; D020125:Mutation, Missense; D055503:Protein Multimerization; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D055815:Young Adult
chemical_list: D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D011506:Proteins; C484746:RSPH1 protein, human; C000589692:RSPH4A protein, human; C573132:RSPH9 protein, human
keywords: cilia; human radial spoke protein 1; human radial spoke protein 4A; human radial spoke protein 9; primary ciliary dyskinesia