All Title Author
Keywords Abstract

PLOS ONE  2012 

Human Amnion Epithelial Cells Induced to Express Functional Cystic Fibrosis Transmembrane Conductance Regulator

DOI: 10.1371/journal.pone.0046533

Full-Text   Cite this paper   Add to My Lib


Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure for cystic fibrosis. With this in mind, we investigated the ability of human amnion epithelial cells (hAECs) to express functional CFTR. We found that hAECs formed 3-dimensional structures and expressed the CFTR gene and protein after culture in Small Airway Growth Medium (SAGM). We also observed a polarized CFTR distribution on the membrane of hAECs cultured in SAGM, similar to that observed in polarized airway cells in vivo. Further, hAECs induced to express CFTR possessed functional iodide/chloride (I?/Cl?) ion channels that were inhibited by the CFTR-inhibitor CFTR-172, indicating the presence of functional CFTR ion channels. These data suggest that hAECs may be a promising source for the development of a cellular therapy for cystic fibrosis.


[1]  Ferrari S, Geddes DM, Alton EW (2002) Barriers to and new approaches for gene therapy and gene delivery in cystic fibrosis. Advanced Drug Delivery Reviews 54: 1373–1393.
[2]  Conese M, Ascenzioni F, Boyd AC, Coutelle C, De Fino I, et al. (2011) Gene and cell therapy for cystic fibrosis: from bench to bedside. Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society 10 Suppl 2S114–128.
[3]  Rosenecker J, Huth S, Rudolph C (2006) Gene therapy for cystic fibrosis lung disease: current status and future perspectives. Current Opinion in Molecular Therapeutics 8: 439–445.
[4]  Hida K, Lai SK, Suk JS, Won SY, Boyle MP, et al. (2011) Common gene therapy viral vectors do not efficiently penetrate sputum from cystic fibrosis patients. PLoS ONE 6: e19919.
[5]  Atkinson TJ (2008) Cystic fibrosis, vector-mediated gene therapy, and relevance of toll-like receptors: a review of problems, progress, and possibilities. Current Gene Therapy 8: 201–207.
[6]  Griesenbach U, Geddes DM, Alton EW (2004) Advances in cystic fibrosis gene therapy. Current Opinion in Pulmonary Medicine 10: 542–546.
[7]  Griesenbach U, Geddes DM, Alton EW (2004) Gene therapy for cystic fibrosis: an example for lung gene therapy. Gene Therapy 11: S43–50.
[8]  Murphy S, Rosli S, Acharya R, Mathias L, Lim R, et al. (2010) Amnion epithelial cell isolation and characterization for clinical use. Curr Protoc Stem Cell Biol 1: 1E.6.1–1E6.23.
[9]  Ilancheran S, Michalska A, Peh G, Wallace EM, Pera M, et al. (2007) Stem cells derived from human fetal membranes display multilineage differentiation potential. Biology of reproduction 77: 577–588.
[10]  Moodley Y, Ilancheran S, Samuel C, Vaghjiani V, Atienza D, et al. (2010) Human amnion epithelial cell transplantation abrogates lung fibrosis and augments repair. American journal of respiratory and critical care medicine 182: 643–651.
[11]  Uchida M, McDermott G, Wetzler M, Le Gros MA, Myllys M, et al. (2009) Soft X-ray tomography of phenotypic switching and the cellular response to antifungal peptoids in Candida albicans. Proceedings of the National Academy of Sciences of the United States of America 106: 19375–19380.
[12]  Erdogan H, Fessler JA (1999) Ordered subsets algorithms for transmission tomography. Physics in Medicine and Biology 44: 2835–2851.
[13]  Stayman JW, Fessler JA (2000) Regularization for uniform spatial resolution properties in penalized-likelihood image reconstruction. IEEE Transactions on Medical Imaging 19: 601–615.
[14]  Tang W, Wildey MJ (2004) Development of a colorimetric method for functional chloride channel assay. Journal of Biomolecular Screening 9: 607–613.
[15]  Maduke M, Miller C, Mindell JA (2000) A decade of CLC chloride channels: structure, mechanism, and many unsettled questions. Annual Review of Biophysics and Biomolecular Structure 29: 411–438.
[16]  Paterson D, Jonge MDd, Howard DL, Lewis W, McKinlay J, et al. (2011) The X-ray Fluorescence Microscopy Beamline at the Australian Synchrotron. AIP Conference Proceedings 1365: 219–222.
[17]  Vogt S (2003) MAPS : A set of software tools for analysis and visualization of 3D X-ray fluorescence data sets. J Phys IV France 104: 635–638.
[18]  Paracchini V, Carbone A, Colombo F, Castellani S, Mazzucchelli S, et al. (2012) Amniotic mesenchymal stem cells: a new source for hepatocyte-like cells and induction of CFTR expression by coculture with cystic fibrosis airway epithelial cells. Journal of biomedicine & biotechnology 2012: 575471.
[19]  Chen YT, Li W, Hayashida Y, He H, Chen SY, et al. (2007) Human amniotic epithelial cells as novel feeder layers for promoting ex vivo expansion of limbal epithelial progenitor cells. Stem Cells 25: 1995–2005.
[20]  Kartner N, Augustinas O, Jensen TJ, Naismith AL, Riordan JR (1992) Mislocalization of delta F508 CFTR in cystic fibrosis sweat gland. Nature Genetics 1: 321–327.
[21]  Kreda SM, Mall M, Mengos A, Rochelle L, Yankaskas J, et al. (2005) Characterization of wild-type and deltaF508 cystic fibrosis transmembrane regulator in human respiratory epithelia. Molecular Biology of the Cell 16: 2154–2167.
[22]  Chappe V, Irvine T, Liao J, Evagelidis A, Hanrahan JW (2005) Phosphorylation of CFTR by PKA promotes binding of the regulatory domain. EMBO Journal 24: 2730–2740.
[23]  Lu M, Leng Q, Egan ME, Caplan MJ, Boulpaep EL, et al. (2006) CFTR is required for PKA-regulated ATP sensitivity of Kir1.1 potassium channels in mouse kidney. Journal of Clinical Investigation 116: 797–807.
[24]  Ito Y, Mizuno Y, Aoyama M, Kume H, Yamaki K (2000) CFTR-Mediated anion conductance regulates Na(+)-K(+)-pump activity in Calu-3 human airway cells. Biochemical and Biophysical Research Communications 274: 230–235.
[25]  Walsh DE, Harvey BJ, Urbach V (2000) CFTR regulation of intracellular calcium in normal and cystic fibrosis human airway epithelia. Journal of Membrane Biology 177: 209–219.
[26]  Naren AP, Nelson DJ, Xie W, Jovov B, Pevsner J, et al. (1997) Regulation of CFTR chloride channels by syntaxin and Munc18 isoforms. Nature 390: 302–305.
[27]  Vankeerberghen A, Cuppens H, Cassiman JJ (2002) The cystic fibrosis transmembrane conductance regulator: an intriguing protein with pleiotropic functions. Journal of Cystic Fibrosis 1: 13–29.
[28]  Murphy S, Lim R, Dickinson H, Acharya R, Rosli S, et al. (2011) Human amnion epithelial cells prevent bleomycin-induced lung injury and preserve lung function. Cell transplantation 20: 909–923.


comments powered by Disqus