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Protocadherin-1 localization and cell-adhesion function in airway epithelial cells in asthma

Protocadherin-1 localization and cell-adhesion function in airway epithelial cells in asthma
Protocadherin-1 localization and cell-adhesion function in airway epithelial cells in asthma
Background

The asthma gene PCDH1 encodes Protocadherin-1, a putative adhesion molecule of unknown function expressed in the airway epithelium. Here, we characterize the localization, differential expression, homotypic adhesion specificity and function of PCDH1 in airway epithelial cells in asthma.

Methods

We performed confocal fluorescence microscopy to determine subcellular localization of PCDH1 in 16HBE cells and primary bronchial epithelial cells (PBECs) grown at air-liquid interface. Next, to compare PCDH1 expression and localization in asthma and controls we performed qRT-PCR and fluorescence microscopy in PBECs and immunohistochemistry on airway wall biopsies. We examined homotypic adhesion specificity of HEK293T clones overexpressing fluorescently tagged-PCDH1 isoforms. Finally, to evaluate the role for PCDH1 in epithelial barrier formation and repair, we performed siRNA knockdown-studies and measured epithelial resistance.

Results

PCDH1 localized to the cell membrane at cell-cell contact sites, baso-lateral to adherens junctions, with increasing expression during epithelial differentiation. No differences in gene expression or localization of PCDH1 isoforms expressing the extracellular domain were observed in either PBECs or airway wall biopsies between asthma patients and controls. Overexpression of PCDH1 mediated homotypic interaction, whereas downregulation of PCDH1 reduced epithelial barrier formation, and impaired repair after wounding.

Conclusions

In conclusion, PCDH1 is localized to the cell membrane of bronchial epithelial cells baso-lateral to the adherens junction. Expression of PCDH1 is not reduced nor delocalized in asthma even though PCDH1 contributes to homotypic adhesion, epithelial barrier formation and repair.
1932-6203
1-21
Faura Tellez, Grissel
a4dcdc41-af1d-444a-b6c4-14f4efd1a93c
Willemse, Brigitte W.M.
bf59c206-763b-40e1-b5de-692e80506cf2
Brouwer, Uilke
260b1582-747d-48bf-9ca5-a16bdd9fda56
Nijboer-Brinksma, Susan
a9490658-1626-432d-b3b4-ac233134c1d1
Vandepoele, Karl
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Noordhoek, Jacobien A.
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Heijink, Irene
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de Vries, Maaike
b924ba42-51fc-47e9-ae19-fa61424c6fc7
Smithers, Natalie P.
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Postma, Dirkje S.
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Timens, Wim
2610a41f-0c54-458b-a8c3-ae9cfb2c5e21
Wiffen, Laura
d49c6ed3-a7cb-4bb7-bf41-2ee142e590a0
van Roy, Frans
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Holloway, John W
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Lackie, Peter M
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Nawijn, Martijn C.
16a331eb-87b5-48e7-976f-ddbb3fc9efe9
Koppelman, Gerard H.
8d04aab8-5795-4e62-b64a-de7e368c0d1d
Faura Tellez, Grissel
a4dcdc41-af1d-444a-b6c4-14f4efd1a93c
Willemse, Brigitte W.M.
bf59c206-763b-40e1-b5de-692e80506cf2
Brouwer, Uilke
260b1582-747d-48bf-9ca5-a16bdd9fda56
Nijboer-Brinksma, Susan
a9490658-1626-432d-b3b4-ac233134c1d1
Vandepoele, Karl
56deacff-22d8-469d-b3ab-d4a659a943aa
Noordhoek, Jacobien A.
b2d9c4ed-f2cf-4cfc-8ef7-f08e08a0d29c
Heijink, Irene
3476c6ad-4046-4170-92c0-1451c9cf8029
de Vries, Maaike
b924ba42-51fc-47e9-ae19-fa61424c6fc7
Smithers, Natalie P.
debb6cb4-c1c7-4f8c-93bc-124444f7d11c
Postma, Dirkje S.
23c1567d-a264-48a1-9ab4-01beda374e42
Timens, Wim
2610a41f-0c54-458b-a8c3-ae9cfb2c5e21
Wiffen, Laura
d49c6ed3-a7cb-4bb7-bf41-2ee142e590a0
van Roy, Frans
0c236c08-cfcb-4231-838b-d42162757f93
Holloway, John W
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Lackie, Peter M
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Nawijn, Martijn C.
16a331eb-87b5-48e7-976f-ddbb3fc9efe9
Koppelman, Gerard H.
8d04aab8-5795-4e62-b64a-de7e368c0d1d

Faura Tellez, Grissel, Willemse, Brigitte W.M., Brouwer, Uilke, Nijboer-Brinksma, Susan, Vandepoele, Karl, Noordhoek, Jacobien A., Heijink, Irene, de Vries, Maaike, Smithers, Natalie P., Postma, Dirkje S., Timens, Wim, Wiffen, Laura, van Roy, Frans, Holloway, John W, Lackie, Peter M, Nawijn, Martijn C. and Koppelman, Gerard H. (2016) Protocadherin-1 localization and cell-adhesion function in airway epithelial cells in asthma. PLoS ONE, 11 (10), 1-21. (doi:10.1371/journal.pone.0163967). (PMID:27701444)

Record type: Article

Abstract

Background

The asthma gene PCDH1 encodes Protocadherin-1, a putative adhesion molecule of unknown function expressed in the airway epithelium. Here, we characterize the localization, differential expression, homotypic adhesion specificity and function of PCDH1 in airway epithelial cells in asthma.

Methods

We performed confocal fluorescence microscopy to determine subcellular localization of PCDH1 in 16HBE cells and primary bronchial epithelial cells (PBECs) grown at air-liquid interface. Next, to compare PCDH1 expression and localization in asthma and controls we performed qRT-PCR and fluorescence microscopy in PBECs and immunohistochemistry on airway wall biopsies. We examined homotypic adhesion specificity of HEK293T clones overexpressing fluorescently tagged-PCDH1 isoforms. Finally, to evaluate the role for PCDH1 in epithelial barrier formation and repair, we performed siRNA knockdown-studies and measured epithelial resistance.

Results

PCDH1 localized to the cell membrane at cell-cell contact sites, baso-lateral to adherens junctions, with increasing expression during epithelial differentiation. No differences in gene expression or localization of PCDH1 isoforms expressing the extracellular domain were observed in either PBECs or airway wall biopsies between asthma patients and controls. Overexpression of PCDH1 mediated homotypic interaction, whereas downregulation of PCDH1 reduced epithelial barrier formation, and impaired repair after wounding.

Conclusions

In conclusion, PCDH1 is localized to the cell membrane of bronchial epithelial cells baso-lateral to the adherens junction. Expression of PCDH1 is not reduced nor delocalized in asthma even though PCDH1 contributes to homotypic adhesion, epithelial barrier formation and repair.

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More information

Accepted/In Press date: 16 September 2016
e-pub ahead of print date: 4 October 2016
Published date: 4 October 2016
Organisations: Human Development & Health, Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 401165
URI: https://eprints.soton.ac.uk/id/eprint/401165
ISSN: 1932-6203
PURE UUID: 46e34353-1812-4edb-8068-1b2668529f4c
ORCID for John W Holloway: ORCID iD orcid.org/0000-0001-9998-0464
ORCID for Peter M Lackie: ORCID iD orcid.org/0000-0001-7138-3764

Catalogue record

Date deposited: 05 Oct 2016 14:44
Last modified: 26 Nov 2019 02:02

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Contributors

Author: Grissel Faura Tellez
Author: Brigitte W.M. Willemse
Author: Uilke Brouwer
Author: Susan Nijboer-Brinksma
Author: Karl Vandepoele
Author: Jacobien A. Noordhoek
Author: Irene Heijink
Author: Maaike de Vries
Author: Natalie P. Smithers
Author: Dirkje S. Postma
Author: Wim Timens
Author: Laura Wiffen
Author: Frans van Roy
Author: John W Holloway ORCID iD
Author: Peter M Lackie ORCID iD
Author: Martijn C. Nawijn
Author: Gerard H. Koppelman

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