Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells
We previously found that human amniotic mesenchymal stem cells (hAMSCs) in coculture with CF immortalised airway epithelial cells (CFBE41o- line, CFBE) on Transwell® filters acquired an epithelial phenotype and led to the expression of a mature and functional CFTR protein. In order to explore the ro...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2018-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2018/1203717 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832564942088175616 |
|---|---|
| author | Annalucia Carbone Roberto Zefferino Elisa Beccia Valeria Casavola Stefano Castellani Sante Di Gioia Valentina Giannone Manuela Seia Antonella Angiolillo Carla Colombo Maria Favia Massimo Conese |
| author_facet | Annalucia Carbone Roberto Zefferino Elisa Beccia Valeria Casavola Stefano Castellani Sante Di Gioia Valentina Giannone Manuela Seia Antonella Angiolillo Carla Colombo Maria Favia Massimo Conese |
| author_sort | Annalucia Carbone |
| collection | DOAJ |
| description | We previously found that human amniotic mesenchymal stem cells (hAMSCs) in coculture with CF immortalised airway epithelial cells (CFBE41o- line, CFBE) on Transwell® filters acquired an epithelial phenotype and led to the expression of a mature and functional CFTR protein. In order to explore the role of gap junction- (GJ-) mediated intercellular communication (GJIC) in this rescue, cocultures (hAMSC : CFBE, 1 : 5 ratio) were studied for the formation of GJIC, before and after silencing connexin 43 (Cx43), a major component of GJs. Functional GJs in cocultures were inhibited when the expression of the Cx43 protein was downregulated. Transfection of cocultures with siRNA against Cx43 resulted in the absence of specific CFTR signal on the apical membrane and reduction in the mature form of CFTR (band C), and in parallel, the CFTR-dependent chloride channel activity was significantly decreased. Cx43 downregulation determined also a decrease in transepithelial resistance and an increase in paracellular permeability as compared with control cocultures, implying that GJIC may regulate CFTR expression and function that in turn modulate airway epithelium tightness. These results indicate that GJIC is involved in the correction of CFTR chloride channel activity upon the acquisition of an epithelial phenotype by hAMSCs in coculture with CF cells. |
| format | Article |
| id | doaj-art-323aa6810000408bbca5d833b79832f0 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-323aa6810000408bbca5d833b79832f02025-02-03T01:09:46ZengWileyStem Cells International1687-966X1687-96782018-01-01201810.1155/2018/12037171203717Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- CellsAnnalucia Carbone0Roberto Zefferino1Elisa Beccia2Valeria Casavola3Stefano Castellani4Sante Di Gioia5Valentina Giannone6Manuela Seia7Antonella Angiolillo8Carla Colombo9Maria Favia10Massimo Conese11Division of Internal Medicine and Chronobiology Unit, IRCCS “Casa Sollievo della Sofferenza”, San Giovanni Rotondo, Foggia, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyDepartment of Bioscience, Biotechnology and Biopharmaceutics, University of Bari, Bari, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyMedical Genetics Laboratory, Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Milan, ItalyMedical Genetics Laboratory, Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Milan, ItalyDepartment of Medicine and Health Sciences “V. Tiberio”, University of Molise, Campobasso, ItalyCystic Fibrosis Center, Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Department of Pathophysiology and Transplantation, University of Milan, Milan, ItalyDepartment of Bioscience, Biotechnology and Biopharmaceutics, University of Bari, Bari, ItalyDepartment of Medical and Surgical Sciences, University of Foggia, Foggia, ItalyWe previously found that human amniotic mesenchymal stem cells (hAMSCs) in coculture with CF immortalised airway epithelial cells (CFBE41o- line, CFBE) on Transwell® filters acquired an epithelial phenotype and led to the expression of a mature and functional CFTR protein. In order to explore the role of gap junction- (GJ-) mediated intercellular communication (GJIC) in this rescue, cocultures (hAMSC : CFBE, 1 : 5 ratio) were studied for the formation of GJIC, before and after silencing connexin 43 (Cx43), a major component of GJs. Functional GJs in cocultures were inhibited when the expression of the Cx43 protein was downregulated. Transfection of cocultures with siRNA against Cx43 resulted in the absence of specific CFTR signal on the apical membrane and reduction in the mature form of CFTR (band C), and in parallel, the CFTR-dependent chloride channel activity was significantly decreased. Cx43 downregulation determined also a decrease in transepithelial resistance and an increase in paracellular permeability as compared with control cocultures, implying that GJIC may regulate CFTR expression and function that in turn modulate airway epithelium tightness. These results indicate that GJIC is involved in the correction of CFTR chloride channel activity upon the acquisition of an epithelial phenotype by hAMSCs in coculture with CF cells.http://dx.doi.org/10.1155/2018/1203717 |
| spellingShingle | Annalucia Carbone Roberto Zefferino Elisa Beccia Valeria Casavola Stefano Castellani Sante Di Gioia Valentina Giannone Manuela Seia Antonella Angiolillo Carla Colombo Maria Favia Massimo Conese Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells Stem Cells International |
| title | Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells |
| title_full | Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells |
| title_fullStr | Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells |
| title_full_unstemmed | Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells |
| title_short | Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells |
| title_sort | gap junctions are involved in the rescue of cftr dependent chloride efflux by amniotic mesenchymal stem cells in coculture with cystic fibrosis cfbe41o cells |
| url | http://dx.doi.org/10.1155/2018/1203717 |
| work_keys_str_mv | AT annaluciacarbone gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT robertozefferino gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT elisabeccia gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT valeriacasavola gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT stefanocastellani gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT santedigioia gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT valentinagiannone gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT manuelaseia gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT antonellaangiolillo gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT carlacolombo gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT mariafavia gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells AT massimoconese gapjunctionsareinvolvedintherescueofcftrdependentchlorideeffluxbyamnioticmesenchymalstemcellsincoculturewithcysticfibrosiscfbe41ocells |