title: Evaluation of genome-wide expression profiles of blood and sputum neutrophils in cystic fibrosis patients before and after antibiotic therapy.
abstract: In seeking more specific biomarkers of the cystic fibrosis (CF) lung inflammatory disease that would be sensitive to antibiotic therapy, we sought to evaluate the gene expression profiles of neutrophils in CF patients before treatment in comparison with non-CF healthy individuals and after antibiotic treatment. Genes involved in neutrophil-mediated inflammation, i.e. chemotaxis, respiratory burst, apoptosis, and granule exocytosis, were the targets of this study. Microarray analysis was carried out in blood and airway neutrophils from CF patients and in control subjects. A fold change (log) threshold of 1.4 and a cut-off of p<0.05 were utilized to identify significant genes. Community networks and principal component analysis were used to distinguish the groups of controls, pre- and post-therapy patients. Control subjects and CF patients before therapy were readily separated, whereas a clear distinction between patients before and after antibiotic therapy was not possible. Blood neutrophils before therapy presented 269 genes down-regulated and 56 up-regulated as compared with control subjects. Comparison between the same patients before and after therapy showed instead 44 genes down-regulated and 72 up-regulated. Three genes appeared to be sensitive to therapy and returned to "healthy" condition: phorbol-12-myristate-13-acetate-induced protein 1 (PMAIP1), hydrogen voltage-gated channel 1 (HVCN1), and β-arrestin 1 (ARRB1). The up-regulation of these genes after therapy were confirmed by real time PCR. In airway neutrophils, 1029 genes were differentially expressed post- vs pre-therapy. Of these, 30 genes were up-regulated and 75 down-regulated following antibiotic treatment. However, biological plausibility determined that only down-regulated genes belonged to the gene classes studied for blood neutrophils. Finally, it was observed that commonly expressed genes showed a greater variability in airway neutrophils than that found in blood neutrophils, both before and after therapy. These results indicate more specific targets for future interventions in CF patients involving respiratory burst, apoptosis, and granule exocytosis.
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Conese
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Castellani
authors.affiliation: Laboratory of Preclinical and Translational Research, IRCCS-CROB, Rionero in Vulture, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Lepore
authors.affiliation: Medical Genetics Unit, IRCCS "Casa Sollievo della Sofferenza (CSS)" Hospital, San Giovanni Rotondo, Italy.
authors.forename: Orazio
authors.initials: O
authors.lastname: Palumbo
authors.affiliation: Centro di Riferimento Regionale Pugliese per la Fibrosi Cistica, Az. Univ. Osp. "Policlinico", Bari, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Manca
authors.affiliation: Department of Biomedical Sciences and Human Oncology, Section of Pediatrics, University of Bari, Bari, Italy.
authors.forename: Teresa
authors.initials: T
authors.lastname: Santostasi
authors.affiliation: Department of Biomedical Sciences and Human Oncology, Section of Pediatrics, University of Bari, Bari, Italy.
authors.forename: Angela Maria
authors.initials: AM
authors.lastname: Polizzi
authors.affiliation: Biostatistic Unit, IRCCS "Casa Sollievo della Sofferenza (CSS)" Hospital, San Giovanni Rotondo, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Copetti
authors.affiliation: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Sante
authors.initials: S
authors.lastname: Di Gioia
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Casavola
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Guerra
authors.affiliation: Centro di Riferimento Regionale Pugliese per la Fibrosi Cistica, Az. Univ. Osp. "Policlinico", Bari, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Diana
authors.affiliation: Department of Biomedical Sciences and Human Oncology, Section of Hygiene, University of Bari, Bari, Italy.
authors.forename: Pasqualina
authors.initials: P
authors.lastname: Montemurro
authors.affiliation: Department of Biomedical Sciences and Human Oncology, Section of Hygiene, University of Bari, Bari, Italy.
authors.forename: Maria Addolorata
authors.initials: MA
authors.lastname: Mariggiò
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Crescenzio
authors.initials: C
authors.lastname: Gallo
authors.affiliation: Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
authors.forename: Angela Bruna
authors.initials: AB
authors.lastname: Maffione
authors.affiliation: Medical Genetics Unit, IRCCS "Casa Sollievo della Sofferenza (CSS)" Hospital, San Giovanni Rotondo, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Carella
mesh_terms: D000293:Adolescent; D000328:Adult; D000900:Anti-Bacterial Agents / Q000494:pharmacology / Q000627:therapeutic use*; D016022:Case-Control Studies; D002648:Child; D003550:Cystic Fibrosis / Q000097:blood* / Q000188:drug therapy / Q000235:genetics* / Q000503:physiopathology; D003710:Demography; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D015894:Genome, Human; D006801:Humans; D007249:Inflammation / Q000097:blood; D008168:Lung / Q000187:drug effects / Q000473:pathology; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012129:Respiratory Function Tests; D013183:Sputum / Q000166:cytology* / Q000382:microbiology; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D055815:Young Adult
chemical_list: D000900:Anti-Bacterial Agents
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