Please use this identifier to cite or link to this item: http://hdl.handle.net/2381/36837
Title: Targeted NGS gene panel identifies mutations in RSPH1 causing primary ciliary dyskinesia and a common mechanism for ciliary central pair agenesis due to radial spoke defects
Authors: Onoufriadis, A.
Shoemark, A.
Schmidts, M.
Patel, M.
Jimenez, G.
Liu, H.
Thomas, Biju
Dixon, M.
Hirst, Robert Anthony
Rutman, Andrew
Burgoyne, T.
Williams, Christopher
Scully, J.
Bolard, F.
Lafitte, J. J.
Beales, P. L.
Hogg, C.
Yang, P.
Chung, E. M.
Emes, R. D.
O'Callaghan, Christopher
UK10K
Bouvagnet, P.
Mitchison, H. M.
First Published: 11-Feb-2014
Publisher: Oxford University Press (OUP)
Citation: Human Molecular Genetics, 2014, 23 (13), pp. 3362-3374
Abstract: Primary ciliary dyskinesia (PCD) is an inherited chronic respiratory obstructive disease with randomized body laterality and infertility, resulting from cilia and sperm dysmotility. PCD is characterized by clinical variability and extensive genetic heterogeneity, associated with different cilia ultrastructural defects and mutations identified in >20 genes. Next generation sequencing (NGS) technologies therefore present a promising approach for genetic diagnosis which is not yet in routine use. We developed a targeted panel-based NGS pipeline to identify mutations by sequencing of selected candidate genes in 70 genetically undefined PCD patients. This detected loss-of-function RSPH1 mutations in four individuals with isolated central pair (CP) agenesis and normal body laterality, from two unrelated families. Ultrastructural analysis in RSPH1-mutated cilia revealed transposition of peripheral outer microtubules into the 'empty' CP space, accompanied by a distinctive intermittent loss of the central pair microtubules. We find that mutations in RSPH1, RSPH4A and RSPH9, which all encode homologs of components of the 'head' structure of ciliary radial spoke complexes identified in Chlamydomonas, cause clinical phenotypes that appear to be indistinguishable except at the gene level. By high-resolution immunofluorescence we identified a loss of RSPH4A and RSPH9 along with RSPH1 from RSPH1-mutated cilia, suggesting RSPH1 mutations may result in loss of the entire spoke head structure. CP loss is seen in up to 28% of PCD cases, in whom laterality determination specified by CP-less embryonic node cilia remains undisturbed. We propose this defect could arise from instability or agenesis of the ciliary central microtubules due to loss of their normal radial spoke head tethering.
DOI Link: 10.1093/hmg/ddu046
ISSN: 0964-6906
eISSN: 1460-2083
Links: http://hmg.oxfordjournals.org/content/23/13/3362
http://hdl.handle.net/2381/36837
Version: Publisher Version
Status: Peer-reviewed
Type: Journal Article
Rights: Copyright © The Author 2014. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Appears in Collections:Published Articles, Dept. of Infection, Immunity and Inflammation



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