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A quantitative super-resolution imaging toolbox for diagnosis of motile ciliopathies

A quantitative super-resolution imaging toolbox for diagnosis of motile ciliopathies
A quantitative super-resolution imaging toolbox for diagnosis of motile ciliopathies
Airway clearance of pathogens and particulates relies on motile cilia. Impaired cilia motility can lead to reduction in lung function, lung transplant, or death in some cases. More than 50 proteins regulating cilia motility are linked to primary ciliary dyskinesia (PCD), a heterogeneous, mainly recessive genetic lung disease. Accurate PCD molecular diagnosis is essential for identifying therapeutic targets and for initiating therapies that can stabilize lung function, thereby reducing socioeconomic impact of the disease. To date, PCD diagnosis has mainly relied on nonquantitative methods that have limited sensitivity or require a priori knowledge of the genes involved. Here, we developed a quantitative super-resolution microscopy workflow: (i) to increase sensitivity and throughput, (ii) to detect structural defects in PCD patients’ cells, and (iii) to quantify motility defects caused by yet to be found PCD genes. Toward these goals, we built a localization map of PCD proteins by three-dimensional structured illumination microscopy and implemented quantitative image analysis and machine learning to detect protein mislocalization, we analyzed axonemal structure by stochastic optical reconstruction microscopy, and we developed a high-throughput method for detecting motile cilia uncoordination by rotational polarity. Together, our data show that super-resolution methods are powerful tools for improving diagnosis of motile ciliopathies.
1946-6234
1-13
Liu, Zhen
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Nguyen, Quynh P.H.
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Guan, Qingxu
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Albulescu, Alexandra
7cedc7ca-e39e-492d-b21e-40d3b50957fb
Erdman, Lauren
56f59482-a9ec-4fa7-b6a9-2bdd0e81c49f
Mahdaviyeh, Yasaman
1d4cd0fb-f868-4c72-95e7-69f49647b7b9
Kang, Jasmine
408a8804-e77d-45dd-b17a-76ae30223b04
Ouyang, Hong
5f9e3b35-6214-48d0-ab6c-6c003421d68c
Hegele, Richard G.
621d2476-e7ba-4da7-99de-e57d5bdb2664
Moraes, Theo
6389bdc2-15d8-4ee6-bed7-39822845d4ef
Goldenberg, Anna
0c348af0-1f1d-4f08-81f9-f18b1893d82f
Del, Sharon D.
d3c4dba7-be9c-45f0-8b0b-0f9397466e46
Mennella, Vito
43c60e29-c0a7-4ab8-8e5c-fcb59f70a28a
Liu, Zhen
df75de79-df16-4f2a-ab45-6897233b6860
Nguyen, Quynh P.H.
92606ace-94f7-411c-895f-ba3e404c8dd2
Guan, Qingxu
d43a8a55-70cf-410c-9988-9e028cd2127c
Albulescu, Alexandra
7cedc7ca-e39e-492d-b21e-40d3b50957fb
Erdman, Lauren
56f59482-a9ec-4fa7-b6a9-2bdd0e81c49f
Mahdaviyeh, Yasaman
1d4cd0fb-f868-4c72-95e7-69f49647b7b9
Kang, Jasmine
408a8804-e77d-45dd-b17a-76ae30223b04
Ouyang, Hong
5f9e3b35-6214-48d0-ab6c-6c003421d68c
Hegele, Richard G.
621d2476-e7ba-4da7-99de-e57d5bdb2664
Moraes, Theo
6389bdc2-15d8-4ee6-bed7-39822845d4ef
Goldenberg, Anna
0c348af0-1f1d-4f08-81f9-f18b1893d82f
Del, Sharon D.
d3c4dba7-be9c-45f0-8b0b-0f9397466e46
Mennella, Vito
43c60e29-c0a7-4ab8-8e5c-fcb59f70a28a

Liu, Zhen, Nguyen, Quynh P.H., Guan, Qingxu, Albulescu, Alexandra, Erdman, Lauren, Mahdaviyeh, Yasaman, Kang, Jasmine, Ouyang, Hong, Hegele, Richard G., Moraes, Theo, Goldenberg, Anna, Del, Sharon D. and Mennella, Vito (2020) A quantitative super-resolution imaging toolbox for diagnosis of motile ciliopathies. Science Translational Medicine, 12 (535), 1-13, [eaay0071]. (doi:10.1126/scitranslmed.aay0071).

Record type: Article

Abstract

Airway clearance of pathogens and particulates relies on motile cilia. Impaired cilia motility can lead to reduction in lung function, lung transplant, or death in some cases. More than 50 proteins regulating cilia motility are linked to primary ciliary dyskinesia (PCD), a heterogeneous, mainly recessive genetic lung disease. Accurate PCD molecular diagnosis is essential for identifying therapeutic targets and for initiating therapies that can stabilize lung function, thereby reducing socioeconomic impact of the disease. To date, PCD diagnosis has mainly relied on nonquantitative methods that have limited sensitivity or require a priori knowledge of the genes involved. Here, we developed a quantitative super-resolution microscopy workflow: (i) to increase sensitivity and throughput, (ii) to detect structural defects in PCD patients’ cells, and (iii) to quantify motility defects caused by yet to be found PCD genes. Toward these goals, we built a localization map of PCD proteins by three-dimensional structured illumination microscopy and implemented quantitative image analysis and machine learning to detect protein mislocalization, we analyzed axonemal structure by stochastic optical reconstruction microscopy, and we developed a high-throughput method for detecting motile cilia uncoordination by rotational polarity. Together, our data show that super-resolution methods are powerful tools for improving diagnosis of motile ciliopathies.

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More information

Accepted/In Press date: 28 February 2020
e-pub ahead of print date: 18 March 2020
Published date: 18 March 2020
Additional Information: Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Identifiers

Local EPrints ID: 438783
URI: http://eprints.soton.ac.uk/id/eprint/438783
ISSN: 1946-6234
PURE UUID: 8510f47c-19e6-49ef-ace9-880660f300e4
ORCID for Vito Mennella: ORCID iD orcid.org/0000-0002-4842-9012

Catalogue record

Date deposited: 24 Mar 2020 17:31
Last modified: 05 Jun 2024 18:29

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Contributors

Author: Zhen Liu
Author: Quynh P.H. Nguyen
Author: Qingxu Guan
Author: Alexandra Albulescu
Author: Lauren Erdman
Author: Yasaman Mahdaviyeh
Author: Jasmine Kang
Author: Hong Ouyang
Author: Richard G. Hegele
Author: Theo Moraes
Author: Anna Goldenberg
Author: Sharon D. Del
Author: Vito Mennella ORCID iD

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