Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells

Overview

Journal Stem Cells Int

Publisher Wiley

Specialty Cell Biology

Date 2018 Mar 14

PMID 29531530

Citations 8

Authors

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

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Stem Cell-Based Therapeutic Approaches in Genetic Diseases.

Aslan A, Ari Yuka S Adv Exp Med Biol. 2023; 1436:19-53.

PMID: 36735185 DOI: 10.1007/5584_2023_761.

Phenotypic Alteration of an Established Human Airway Cell Line by Media Selection.

Livnat G, Meeker J, Ostmann A, Strecker L, Clancy J, Brewington J Int J Mol Sci. 2023; 24(2).

PMID: 36674762 PMC: 9862772. DOI: 10.3390/ijms24021246.

Human Amniotic Mesenchymal Stem Cells and Fibroblasts Accelerate Wound Repair of Cystic Fibrosis Epithelium.

Beccia E, Daniello V, Laselva O, Leccese G, Mangiacotti M, Di Gioia S Life (Basel). 2022; 12(5).

PMID: 35629422 PMC: 9144497. DOI: 10.3390/life12050756.

Making Connections: Mesenchymal Stem Cells Manifold Ways to Interact with Neurons.

Tarasiuk O, Ballarini E, Donzelli E, Rodriguez-Menendez V, Bossi M, Cavaletti G Int J Mol Sci. 2022; 23(10).

PMID: 35628600 PMC: 9146463. DOI: 10.3390/ijms23105791.

Pathophysiology of Lung Disease and Wound Repair in Cystic Fibrosis.

Conese M, Di Gioia S Pathophysiology. 2022; 28(1):155-188.

PMID: 35366275 PMC: 8830450. DOI: 10.3390/pathophysiology28010011.

References

Ramachandran S, Karp P, Osterhaus S, Jiang P, Wohlford-Lenane C, Lennox K . Post-transcriptional regulation of cystic fibrosis transmembrane conductance regulator expression and function by microRNAs. Am J Respir Cell Mol Biol. 2013; 49(4):544-51. PMC: 3824042. DOI: 10.1165/rcmb.2012-0430OC. View

Bou Saab J, Losa D, Chanson M, Ruez R . Connexins in respiratory and gastrointestinal mucosal immunity. FEBS Lett. 2014; 588(8):1288-96. DOI: 10.1016/j.febslet.2014.02.059. View

Mall M, Bleich M, Greger R, Schreiber R, Kunzelmann K . The amiloride-inhibitable Na+ conductance is reduced by the cystic fibrosis transmembrane conductance regulator in normal but not in cystic fibrosis airways. J Clin Invest. 1998; 102(1):15-21. PMC: 509060. DOI: 10.1172/JCI2729. View

Ramachandran S, Karp P, Jiang P, Ostedgaard L, Walz A, Fisher J . A microRNA network regulates expression and biosynthesis of wild-type and DeltaF508 mutant cystic fibrosis transmembrane conductance regulator. Proc Natl Acad Sci U S A. 2012; 109(33):13362-7. PMC: 3421220. DOI: 10.1073/pnas.1210906109. View

Favia M, Guerra L, Fanelli T, Cardone R, Monterisi S, Di Sole F . Na+/H+ exchanger regulatory factor 1 overexpression-dependent increase of cytoskeleton organization is fundamental in the rescue of F508del cystic fibrosis transmembrane conductance regulator in human airway CFBE41o- cells. Mol Biol Cell. 2009; 21(1):73-86. PMC: 2801722. DOI: 10.1091/mbc.e09-03-0185. View

Sinclair K, Yerkovich S, Hopkins P, Chambers D . Characterization of intercellular communication and mitochondrial donation by mesenchymal stromal cells derived from the human lung. Stem Cell Res Ther. 2016; 7(1):91. PMC: 4942965. DOI: 10.1186/s13287-016-0354-8. View

Molina S, Stauffer B, Moriarty H, Kim A, McCarty N, Koval M . Junctional abnormalities in human airway epithelial cells expressing F508del CFTR. Am J Physiol Lung Cell Mol Physiol. 2015; 309(5):L475-87. PMC: 4556929. DOI: 10.1152/ajplung.00060.2015. View

Scheckenbach K, Losa D, Dudez T, Bacchetta M, OGrady S, Crespin S . Prostaglandin E₂regulation of cystic fibrosis transmembrane conductance regulator activity and airway surface liquid volume requires gap junctional communication. Am J Respir Cell Mol Biol. 2010; 44(1):74-82. PMC: 3028258. DOI: 10.1165/rcmb.2009-0361OC. View

Stoltz D, Meyerholz D, Welsh M . Origins of cystic fibrosis lung disease. N Engl J Med. 2015; 372(16):1574-5. DOI: 10.1056/NEJMc1502191. View

10.

Miki T, Strom S . Amnion-derived pluripotent/multipotent stem cells. Stem Cell Rev. 2007; 2(2):133-42. DOI: 10.1007/s12015-006-0020-0. View

11.

Hayes M, Curley G, Ansari B, Laffey J . Clinical review: Stem cell therapies for acute lung injury/acute respiratory distress syndrome - hope or hype?. Crit Care. 2012; 16(2):205. PMC: 3681334. DOI: 10.1186/cc10570. View

12.

Ruch R, Porter S, Koffler L, Dwyer-Nield L, Malkinson A . Defective gap junctional intercellular communication in lung cancer: loss of an important mediator of tissue homeostasis and phenotypic regulation. Exp Lung Res. 2001; 27(3):231-43. DOI: 10.1080/019021401300053984. View

13.

Whitsett J, Alenghat T . Respiratory epithelial cells orchestrate pulmonary innate immunity. Nat Immunol. 2014; 16(1):27-35. PMC: 4318521. DOI: 10.1038/ni.3045. View

14.

Meister G, Tuschl T . Mechanisms of gene silencing by double-stranded RNA. Nature. 2004; 431(7006):343-9. DOI: 10.1038/nature02873. View

15.

Weiss D, Chambers D, Giangreco A, Keating A, Kotton D, Lelkes P . An official American Thoracic Society workshop report: stem cells and cell therapies in lung biology and diseases. Ann Am Thorac Soc. 2015; 12(4):S79-97. PMC: 5466184. DOI: 10.1513/AnnalsATS.201502-086ST. View

16.

Kean T, Lin P, Caplan A, Dennis J . MSCs: Delivery Routes and Engraftment, Cell-Targeting Strategies, and Immune Modulation. Stem Cells Int. 2013; 2013:732742. PMC: 3755386. DOI: 10.1155/2013/732742. View

17.

Roomans G . Tissue engineering and the use of stem/progenitor cells for airway epithelium repair. Eur Cell Mater. 2010; 19:284-99. DOI: 10.22203/ecm.v019a27. View

18.

Oglesby I, Chotirmall S, McElvaney N, M Greene C . Regulation of cystic fibrosis transmembrane conductance regulator by microRNA-145, -223, and -494 is altered in ΔF508 cystic fibrosis airway epithelium. J Immunol. 2013; 190(7):3354-62. DOI: 10.4049/jimmunol.1202960. View

19.

Lesimple P, Liao J, Robert R, Gruenert D, Hanrahan J . Cystic fibrosis transmembrane conductance regulator trafficking modulates the barrier function of airway epithelial cell monolayers. J Physiol. 2010; 588(Pt 8):1195-209. PMC: 2872727. DOI: 10.1113/jphysiol.2009.182246. View

20.

Gruenert D, Willems M, Cassiman J, Frizzell R . Established cell lines used in cystic fibrosis research. J Cyst Fibros. 2004; 3 Suppl 2:191-6. DOI: 10.1016/j.jcf.2004.05.040. View