title: Everolimus-induced epithelial to mesenchymal transition (EMT) in bronchial/pulmonary cells: when the dosage does matter in transplantation.
abstract: BACKGROUND
  Everolimus (EVE) is a mammalian target of rapamycin inhibitor (mTOR-I) widely used in transplantation that may determine some severe adverse events, including pulmonary fibrosis. The pathogenic mechanism of mTOR-I-associated pulmonary toxicity is still unclear, but epithelial to mesenchymal transition (EMT) of bronchial/pulmonary cells may play a role.
  
  
  METHODS
  Three cell lines-human type II pneumocyte-derived A549, normal bronchial epithelial, and bronchial epithelial homozygous for the delta F508 cystic fibrosis-causing mutation-were treated with EVE or tacrolimus at different concentrations. Real-time polymerase chain reaction and immunofluorescence were used to evaluate mRNA and protein levels of EMT markers (alpha-SMA, vimentin, fibronectin). Subsequently, in 13 EVE- and 13 tacrolimus-treated patients we compared the rate of lung fibrosis, estimated by an arbitrary pulmonary fibrosis index score (PFIS).
  
  
  RESULTS
  Biomolecular experiments demonstrated that high doses of EVE (100 nM) up-regulated EMT markers in all cell lines at both gene- and protein level. High concentrations of EVE were also able to reduce the mRNA levels of epithelial markers (E-cadherin and ZO-1) and to induce the phosphorylation of AKT. In the in vivo part of the study, PFIS was significantly higher in the EVE-group than the tacrolimus-group (p = 0.03) and correlated with trough levels (R2= 0.35).
  
  
  CONCLUSIONS
  Our data reveal, for the first time, a dose-dependent EVE-induced EMT in airway cells. They suggest that clinicians should employ, wherever possible, low dosages of mTOR-Is in transplant recipients, assessing periodically their pulmonary function.
authors.affiliation: Department of Medicine, Renal Unit, University of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Tomei
authors.affiliation: Department of Medicine, Renal Unit, University of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Masola
authors.affiliation: Department of Medicine, Renal Unit, University of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Granata
authors.affiliation: Department of Medicine, Renal Unit, University of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy.
authors.forename: Gloria
authors.initials: G
authors.lastname: Bellin
authors.affiliation: Institute of Pulmonary Disease, University of Bari, Bari, Italy.
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Carratù
authors.affiliation: Department of Pathology and Diagnostic, Anatomic Pathology Section, University of Verona, Verona, Italy.
authors.forename: Miriam
authors.initials: M
authors.lastname: Ficial
authors.affiliation: Institute of Pulmonary Disease, University of Bari, Bari, Italy.
authors.forename: Valentina Anna
authors.initials: VA
authors.lastname: Ventura
authors.affiliation: Department of Biomedical Sciences, University of Padova, Padua, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Onisto
authors.affiliation: Institute of Pulmonary Disease, University of Bari, Bari, Italy.
authors.forename: Onofrio
authors.initials: O
authors.lastname: Resta
authors.affiliation: Division of Nephrology and Dialysis, School of Medicine Rome, Columbus-Gemelli Hospital Catholic University, Rome, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Gambaro
authors.affiliation: Department of Pathology and Diagnostic, Anatomic Pathology Section, University of Verona, Verona, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Chilosi
authors.affiliation: Department of Medicine, Renal Unit, University of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lupo
authors.affiliation: Department of Medicine, Renal Unit, University of Verona, Piazzale A. Stefani 1, 37126, Verona, VR, Italy. gianluigi.zaza@univr.it.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Zaza
mesh_terms: D000072283:A549 Cells; D000199:Actins / Q000235:genetics / Q000378:metabolism; D000328:Adult; D000368:Aged; D056809:Alveolar Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D001980:Bronchi / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D000068338:Everolimus / Q000009:adverse effects*; D005260:Female; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007166:Immunosuppressive Agents / Q000009:adverse effects*; D008297:Male; D008875:Middle Aged; D016377:Organ Transplantation / Q000009:adverse effects*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000009:adverse effects*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011658:Pulmonary Fibrosis / Q000139:chemically induced* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018570:Risk Assessment; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016559:Tacrolimus / Q000009:adverse effects; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D005353:Fibronectins; D007166:Immunosuppressive Agents; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D014746:Vimentin; D000068338:Everolimus; C546842:MTOR protein, human; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases; D016559:Tacrolimus
keywords: Cystic fibrosis; Epithelial-mesenchymal transition; Everolimus; Fibrosis; Lung; Pulmonary cells; mTOR inhibitor