Primary ciliary dyskinesia with normal ultrastructure: three-dimensional tomography detects absence of DNAH11

Amelia Shoemark (Lead / Corresponding author), Thomas Burgoyne, Robert Kwan, Mellisa Dixon, Mitali P. Patel, Andrew V. Rogers, Alexandros Onoufriadis, Juliet Scully, Farheen Daudvohra, Thomas Cullup, Michael R. Loebinger, Robert Wilson, Eddie M. K. Chung, Andrew Bush, Hannah M. Mitchison, Claire Hogg

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Abstract

In primary ciliary dyskinesia (PCD), motile ciliary dysfunction arises from ciliary defects usually confirmed by transmission electron microscopy (TEM). In 30% of patients, such as those with DNAH11 mutations, apparently normal ultrastructure makes diagnosis difficult. Genetic analysis supports diagnosis, but may not identify definitive causal variants. Electron tomography, an extension of TEM, produces three-dimensional ultrastructural ciliary models with superior resolution to TEM. Our hypothesis is that tomography using existing patient samples will enable visualisation of DNAH11-associated ultrastructural defects. Dual axis tomograms from araldite-embedded nasal cilia were collected in 13 PCD patients with normal ultrastructure (DNAH11 n=7, HYDIN n=2, CCDC65 n=3 and DRC1 n=1) and six healthy controls, then analysed using IMOD and Chimera software.DNAH11 protein is localised to the proximal ciliary region. Within this region, electron tomography indicated a deficiency of >25% of proximal outer dynein arm volume in all patients with DNAH11 mutations (n=7) compared to other patients with PCD and normal ultrastructure (n=6) and healthy controls (n=6). DNAH11 mutations cause a shared abnormality in ciliary ultrastructure previously undetectable by TEM. Advantageously, electron tomography can be used on existing diagnostic samples and establishes a structural abnormality where ultrastructural studies were previously normal.

Original languageEnglish
Article number1701809
JournalEuropean Respiratory Journal
Volume51
Issue number2
Early online date21 Feb 2018
DOIs
Publication statusPublished - Feb 2018

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Kartagener Syndrome
Electron Microscope Tomography
Transmission Electron Microscopy
Tomography
Mutation
Dyneins
Cilia
Nose
Software
Proteins

Cite this

Shoemark, Amelia ; Burgoyne, Thomas ; Kwan, Robert ; Dixon, Mellisa ; Patel, Mitali P. ; Rogers, Andrew V. ; Onoufriadis, Alexandros ; Scully, Juliet ; Daudvohra, Farheen ; Cullup, Thomas ; Loebinger, Michael R. ; Wilson, Robert ; Chung, Eddie M. K. ; Bush, Andrew ; Mitchison, Hannah M. ; Hogg, Claire. / Primary ciliary dyskinesia with normal ultrastructure : three-dimensional tomography detects absence of DNAH11. In: European Respiratory Journal. 2018 ; Vol. 51, No. 2.
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abstract = "In primary ciliary dyskinesia (PCD), motile ciliary dysfunction arises from ciliary defects usually confirmed by transmission electron microscopy (TEM). In 30{\%} of patients, such as those with DNAH11 mutations, apparently normal ultrastructure makes diagnosis difficult. Genetic analysis supports diagnosis, but may not identify definitive causal variants. Electron tomography, an extension of TEM, produces three-dimensional ultrastructural ciliary models with superior resolution to TEM. Our hypothesis is that tomography using existing patient samples will enable visualisation of DNAH11-associated ultrastructural defects. Dual axis tomograms from araldite-embedded nasal cilia were collected in 13 PCD patients with normal ultrastructure (DNAH11 n=7, HYDIN n=2, CCDC65 n=3 and DRC1 n=1) and six healthy controls, then analysed using IMOD and Chimera software.DNAH11 protein is localised to the proximal ciliary region. Within this region, electron tomography indicated a deficiency of >25{\%} of proximal outer dynein arm volume in all patients with DNAH11 mutations (n=7) compared to other patients with PCD and normal ultrastructure (n=6) and healthy controls (n=6). DNAH11 mutations cause a shared abnormality in ciliary ultrastructure previously undetectable by TEM. Advantageously, electron tomography can be used on existing diagnostic samples and establishes a structural abnormality where ultrastructural studies were previously normal.",
author = "Amelia Shoemark and Thomas Burgoyne and Robert Kwan and Mellisa Dixon and Patel, {Mitali P.} and Rogers, {Andrew V.} and Alexandros Onoufriadis and Juliet Scully and Farheen Daudvohra and Thomas Cullup and Loebinger, {Michael R.} and Robert Wilson and Chung, {Eddie M. K.} and Andrew Bush and Mitchison, {Hannah M.} and Claire Hogg",
note = "Support statement: H.M. Mitchison is supported by the Great Ormond Street Hospital Children's Charity, the UK PCD Family Support Group and the National Institute for Health Research (NIHR) Biomedical Research Centre at Great Ormond Street Hospital for Children NHS Foundation Trust and University College London. A. Shoemark is funded by a postdoctoral research fellowship from the NIHR and Health Education England (HEE-HCS-P13-04-004), mentored by C. Hogg, H.M. Mitchison and A. Bush. A. Bush was supported by the NIHR Respiratory Disease Biomedical Research Unit at the Royal Brompton and Harefield NHS Foundation Trust and Imperial College London. This report is independent research arising from a postdoctoral research fellowship supported by the NIHR and Health Education England. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. Funding information for this article has been deposited with the Crossref Funder Registry.",
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Shoemark, A, Burgoyne, T, Kwan, R, Dixon, M, Patel, MP, Rogers, AV, Onoufriadis, A, Scully, J, Daudvohra, F, Cullup, T, Loebinger, MR, Wilson, R, Chung, EMK, Bush, A, Mitchison, HM & Hogg, C 2018, 'Primary ciliary dyskinesia with normal ultrastructure: three-dimensional tomography detects absence of DNAH11', European Respiratory Journal, vol. 51, no. 2, 1701809. https://doi.org/10.1183/13993003.01809-2017

Primary ciliary dyskinesia with normal ultrastructure : three-dimensional tomography detects absence of DNAH11. / Shoemark, Amelia (Lead / Corresponding author); Burgoyne, Thomas; Kwan, Robert; Dixon, Mellisa; Patel, Mitali P.; Rogers, Andrew V.; Onoufriadis, Alexandros; Scully, Juliet; Daudvohra, Farheen; Cullup, Thomas; Loebinger, Michael R.; Wilson, Robert; Chung, Eddie M. K. ; Bush, Andrew; Mitchison, Hannah M.; Hogg, Claire.

In: European Respiratory Journal, Vol. 51, No. 2, 1701809, 02.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Primary ciliary dyskinesia with normal ultrastructure

T2 - three-dimensional tomography detects absence of DNAH11

AU - Shoemark, Amelia

AU - Burgoyne, Thomas

AU - Kwan, Robert

AU - Dixon, Mellisa

AU - Patel, Mitali P.

AU - Rogers, Andrew V.

AU - Onoufriadis, Alexandros

AU - Scully, Juliet

AU - Daudvohra, Farheen

AU - Cullup, Thomas

AU - Loebinger, Michael R.

AU - Wilson, Robert

AU - Chung, Eddie M. K.

AU - Bush, Andrew

AU - Mitchison, Hannah M.

AU - Hogg, Claire

N1 - Support statement: H.M. Mitchison is supported by the Great Ormond Street Hospital Children's Charity, the UK PCD Family Support Group and the National Institute for Health Research (NIHR) Biomedical Research Centre at Great Ormond Street Hospital for Children NHS Foundation Trust and University College London. A. Shoemark is funded by a postdoctoral research fellowship from the NIHR and Health Education England (HEE-HCS-P13-04-004), mentored by C. Hogg, H.M. Mitchison and A. Bush. A. Bush was supported by the NIHR Respiratory Disease Biomedical Research Unit at the Royal Brompton and Harefield NHS Foundation Trust and Imperial College London. This report is independent research arising from a postdoctoral research fellowship supported by the NIHR and Health Education England. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. Funding information for this article has been deposited with the Crossref Funder Registry.

PY - 2018/2

Y1 - 2018/2

N2 - In primary ciliary dyskinesia (PCD), motile ciliary dysfunction arises from ciliary defects usually confirmed by transmission electron microscopy (TEM). In 30% of patients, such as those with DNAH11 mutations, apparently normal ultrastructure makes diagnosis difficult. Genetic analysis supports diagnosis, but may not identify definitive causal variants. Electron tomography, an extension of TEM, produces three-dimensional ultrastructural ciliary models with superior resolution to TEM. Our hypothesis is that tomography using existing patient samples will enable visualisation of DNAH11-associated ultrastructural defects. Dual axis tomograms from araldite-embedded nasal cilia were collected in 13 PCD patients with normal ultrastructure (DNAH11 n=7, HYDIN n=2, CCDC65 n=3 and DRC1 n=1) and six healthy controls, then analysed using IMOD and Chimera software.DNAH11 protein is localised to the proximal ciliary region. Within this region, electron tomography indicated a deficiency of >25% of proximal outer dynein arm volume in all patients with DNAH11 mutations (n=7) compared to other patients with PCD and normal ultrastructure (n=6) and healthy controls (n=6). DNAH11 mutations cause a shared abnormality in ciliary ultrastructure previously undetectable by TEM. Advantageously, electron tomography can be used on existing diagnostic samples and establishes a structural abnormality where ultrastructural studies were previously normal.

AB - In primary ciliary dyskinesia (PCD), motile ciliary dysfunction arises from ciliary defects usually confirmed by transmission electron microscopy (TEM). In 30% of patients, such as those with DNAH11 mutations, apparently normal ultrastructure makes diagnosis difficult. Genetic analysis supports diagnosis, but may not identify definitive causal variants. Electron tomography, an extension of TEM, produces three-dimensional ultrastructural ciliary models with superior resolution to TEM. Our hypothesis is that tomography using existing patient samples will enable visualisation of DNAH11-associated ultrastructural defects. Dual axis tomograms from araldite-embedded nasal cilia were collected in 13 PCD patients with normal ultrastructure (DNAH11 n=7, HYDIN n=2, CCDC65 n=3 and DRC1 n=1) and six healthy controls, then analysed using IMOD and Chimera software.DNAH11 protein is localised to the proximal ciliary region. Within this region, electron tomography indicated a deficiency of >25% of proximal outer dynein arm volume in all patients with DNAH11 mutations (n=7) compared to other patients with PCD and normal ultrastructure (n=6) and healthy controls (n=6). DNAH11 mutations cause a shared abnormality in ciliary ultrastructure previously undetectable by TEM. Advantageously, electron tomography can be used on existing diagnostic samples and establishes a structural abnormality where ultrastructural studies were previously normal.

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U2 - 10.1183/13993003.01809-2017

DO - 10.1183/13993003.01809-2017

M3 - Article

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