Testing human monocytes as a new tool for clinical and preclinical research in CF.

Starting date
September 1, 2011
Duration (months)
24
Managers or local contacts
Angiari Chiara , Sorio Claudio
Keyword
Custic fibrosis, monocytes, biomarker

Functional CFTR is currently directly evaluated in humans using two procedures: rectal biopsies followed by Ussing chamber analysis and nasal potential difference analysis (NPD). Even if only the first require a surgical procedure, nevertheless both can be considered invasive as they require either the removal of tissue (the former) or the use of small electrode on the nasal mucosa (the latter). For this and other practical reasons, for both rectal biopsies and (although to a lesser extent) NPD analysis, the possibility to be repeated multiple times on the same subject, in particular younger subjects, is not practically feasible. Our published data confirm the possibility to apply a well described single-cell fluorescence imaging approach to human monocytes, thus making possible to evaluate CFTR functionality in blood cells. This approach permit to work on with a minimally invasive methodology (a venous blood drawn of 5-10 cc) facilitating the evaluation of CFTR activity in all the cases of relevance in research and clinic, possibly including the evaluation of drugs affecting CFTR function. Our project is the extension of FFC#05/2009 and aims to the evaluation of the monocyte assay on a population of subjects affected by lung disease other than CF and to evaluate ex vivo, on monocyte of CF subjects for the first time, the response to drugs targeting the basic defect of CF, including VX809 and PTC124. It is important to underline the fact that we will have the unique opportunity to evaluate the response to a novel drug, PTC 124, now undergoing a multicentric phase 3 clinical trial in the CF Center in Verona on 15 subjects. Importantly, we have already obtained clearance from the local ethical commettee and from PTC Therapeutics that is organizing this multicenter clinical trial that has recognized the potential of this test for improving the efficacy of future clinical trials. These monocytes will be analyzed also with patch clamp, a highly complex technique that is capable to record the current from single channels present on the cell membrane and represent the gold standard in ion channel research. Eventually, a faster access of the patients to new drugs might be envisaged as this application, once properly validated, might expedite the organization and evaluation of clinical trials based on the use of CFTR correctors.

Sponsors:

Funds: assigned and managed by an external body

Project participants

Chiara Angiari
Mario Rosario Buffelli
Associate Professor
Jan Evert Johansson
Claudio Sorio
Associate Professor
Genny Verze'

Collaboratori esterni

Paola Melotti
AOUI Verona
Giuseppe Bellisola
Azienda Ospedaliera Universitaria Integrata di Verona Patologia Dirigente medico I livello
Baroukh Maurice Assael
AOUI Verona
Publications
Title Authors Year
MudPIT analysis of released proteins in Pseudomonas aeruginosa laboratory and clinical strains in relation to pro-inflammatory effects Bergamini G.; Di Silvestre D.; Mauri P.L.; Cigana C.; Bragonzi A.; De Palma A.; Benazzi L.; Assael B.M.; Melotti P.; Sorio C. 2012
Defective CFTR expression and function are detectable in blood monocytes: development of a new blood test for cystic fibrosis. Sorio, C.; Buffelli, M.; Angiari, C.; Ettorre, M.; Johansson, J.; Vezzalini, M.; Viviani, L.; Ricciardi, M.; Verzè, G.; Assael, B.M.; Melotti, P. 2011
Impact of polymorphism of Multidrug Resistance-associated Protein 1 (ABCC1) gene on the severity of cystic fibrosis. Andrea Mafficini;Myriam Ortombina; Isabelle Sermet-Gaudelius ; Patrick Lebecque; Teresinha Leal; Patrizia Iansa; Gregory Reychler; Karin Dahan; Xaviers Pepermans; Gerard Lenoir; Anissa Leonard; Sorio Claudio; Baroukh Maurice Assael; Paola Melotti 2011
Impact of polymorphism of Multidrug Resistance-associated Protein 1 (ABCC1) gene on the severity of cystic fibrosis Mafficini A, Ortombina M, Sermet-Gaudelius I, Lebecque P, Leal T, Iansa P, Reychler G, Dahan K, Pepermans X, Lenoir G, Leonard A, Sorio C, Assael B, Melotti P 2011
Analysis of CFTR function in human monocytes Sorio C.; Buffelli M.; Ettorre M.; Vezzalini M.; Ricciardi M.; Angiari C.; Johansson J.; Assael B.M.; Melotti P. 2010
Analysis of proteins released from Pseudomonas aeruginosa clinical and laboratory strains: effects of azithromycin G. Bergamini; P. Mauri; C. Cigana; D. Di Silvestre; A. Di Palma; A. Bragonzi; B.M. Assael; C. Sorio; P. Melotti 2009
Azithromycin decreases Glutathione-S-Transferase T1 (GSTT1) and M1 (GSTM1) expression and activity in cystic fibrosis airway epithelial cells Bergamini G. , Cigana C., Sorio C., Della Peruta M., Assael B.M., Melotti P. 2009
Functional Evaluation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) in human monocytes C. Sorio; M. Buffelli; M. Ettorre; M. Vezzalini; M. Ricciardi; C. Angiari; J. Johansson; B.M. Assael; P. Melotti 2009
Impact of the polymorphic variant 5 FR/G-260C in the Multidrug Resistance-associate Protein-1 gene on severity of cystic fibrosis A. Mafficini; M. Ortombina; I. Sermet-Gaudelus; P. Lebecque; T. Leal; A. Leonard; G. Reychler; K. Dahan; X. Papermans; G. Lenoir; C. Sorio; P. Iansa; B.M. Assael; P. Melotti 2009
Analysis of Pseudomonas Aeuginosa secretome: consequences of oxygen limitation in clinical and laboratory strains C. Cigana; G. Bergamini; P. Mauri; C. Sorio; D. Di Silvestre; A. Bragonzi; B.M. Assael; P. Melotti 2008
Azithromicin decreases elevated glutathione-S-transferase T1 (GSTT1) and M1 (GSTM1) expression and activity in cystic fibrosis airways epithelial cells C. Cigana; G. Bergamini; C. Sorio; M. Della Peruta; B.M. Assael; P. Melotti 2008
Development of FT-IR microspectroscopy for pre-clinical and clinical applications Bellisola G.; Bergamini G. ; Bolomini-Vittori M.; Cestelli Guidi M.; Cinque G.; Della Peruta M. ; Giagulli C.; Laudanna C.; Mafficini A.; Marcelli A.; Melotti P.; Piccinini M.; Sorio C. 2008
Proteins released from Pseudomonas aeruginosa (Pa) referent and clinical strains under microaerobiosis and their effects on cystic fibrosis airways Cigana C.; Mauri P.; Di Silvestre D.; Bergamini G.; Bragonzi A.; Assael B.M.; Sorio C.; Melotti P. 2008
Azithromycin (AZM) decreases Glutathione-S-Transferase (GST) - T1 expression and activity in CF airway epithelial cells G. Bergamini; C. Cigana; M. Della Peruta; C. Sorio; B.M. Assael; P. Melotti 2007
Azithromycin (AZM) decreases Glutathione-S-Transferase (GST) -T1 expression and activity in CF airway epithelial cells. Bergamini G. ; Cigana C.; Sorio C.; Della Peruta M.; Assael BM; Melotti P. 2007
Effects of azithromycin on the expression of ATP binding cassette transporters in epithelial cells from the airways of cystic fibrosis patients. C. Cigana; E. Nicolis; M. Pasetto; B. M. Assael; P. Melotti 2007
Relevance of oxygen limitation on Pseudomonas Aeruginosa strains: effects on reelsed protein expression Cigana C.; Bergamini G.; Mauri P.; Sorio C.; Di Silvestre D.; De Palma A.; Bennazzi L.; Bragonzi A.; Assael B.M.; Melotti P. 2007

Activities

Research facilities