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.
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 ultr...
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| Format: | Article |
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Oxford University Press
2014
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| Online Access: | https://eprints.nottingham.ac.uk/34237/ |
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| author | Onoufriadis, Alexandros Shoemark, Amelia Schmidts, Miriam Patel, Mitali Jimenez, Gina Liu, Hui Thomas, Biju Dixon, Melissa Hirst, Robert A. Rutman, Andrew Burgoyne, Thomas Williams, Christopher Scully, Juliet Bolard, Florence Lafitte, Jean-Jacques Beales, Philip L. Hogg, Claire Yang, Pinfen Chung, Eddie M.K. Emes, Richard D. O'Callaghan, Christopher Bouvagnet, Patrice Mitchison, Hannah M. |
| author_facet | Onoufriadis, Alexandros Shoemark, Amelia Schmidts, Miriam Patel, Mitali Jimenez, Gina Liu, Hui Thomas, Biju Dixon, Melissa Hirst, Robert A. Rutman, Andrew Burgoyne, Thomas Williams, Christopher Scully, Juliet Bolard, Florence Lafitte, Jean-Jacques Beales, Philip L. Hogg, Claire Yang, Pinfen Chung, Eddie M.K. Emes, Richard D. O'Callaghan, Christopher Bouvagnet, Patrice Mitchison, Hannah M. |
| author_sort | Onoufriadis, Alexandros |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:22:01Z |
| format | Article |
| id | nottingham-34237 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:22:01Z |
| publishDate | 2014 |
| publisher | Oxford University Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-342372020-05-04T16:43:15Z https://eprints.nottingham.ac.uk/34237/ 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. Onoufriadis, Alexandros Shoemark, Amelia Schmidts, Miriam Patel, Mitali Jimenez, Gina Liu, Hui Thomas, Biju Dixon, Melissa Hirst, Robert A. Rutman, Andrew Burgoyne, Thomas Williams, Christopher Scully, Juliet Bolard, Florence Lafitte, Jean-Jacques Beales, Philip L. Hogg, Claire Yang, Pinfen Chung, Eddie M.K. Emes, Richard D. O'Callaghan, Christopher Bouvagnet, Patrice Mitchison, Hannah M. 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. Oxford University Press 2014-02-11 Article PeerReviewed Onoufriadis, Alexandros, Shoemark, Amelia, Schmidts, Miriam, Patel, Mitali, Jimenez, Gina, Liu, Hui, Thomas, Biju, Dixon, Melissa, Hirst, Robert A., Rutman, Andrew, Burgoyne, Thomas, Williams, Christopher, Scully, Juliet, Bolard, Florence, Lafitte, Jean-Jacques, Beales, Philip L., Hogg, Claire, Yang, Pinfen, Chung, Eddie M.K., Emes, Richard D., O'Callaghan, Christopher, Bouvagnet, Patrice and Mitchison, Hannah M. (2014) 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. Human Molecular Genetics, 23 (13). pp. 3362-3374. ISSN 1460-2083 http://hmg.oxfordjournals.org/content/23/13/3362.long doi:10.1093/hmg/ddu046 doi:10.1093/hmg/ddu046 |
| spellingShingle | Onoufriadis, Alexandros Shoemark, Amelia Schmidts, Miriam Patel, Mitali Jimenez, Gina Liu, Hui Thomas, Biju Dixon, Melissa Hirst, Robert A. Rutman, Andrew Burgoyne, Thomas Williams, Christopher Scully, Juliet Bolard, Florence Lafitte, Jean-Jacques Beales, Philip L. Hogg, Claire Yang, Pinfen Chung, Eddie M.K. Emes, Richard D. O'Callaghan, Christopher Bouvagnet, Patrice Mitchison, Hannah M. 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. |
| 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. |
| title_full | 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. |
| title_fullStr | 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. |
| title_full_unstemmed | 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. |
| title_short | 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. |
| title_sort | 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. |
| url | https://eprints.nottingham.ac.uk/34237/ https://eprints.nottingham.ac.uk/34237/ https://eprints.nottingham.ac.uk/34237/ |