High doses of Everolimus induce transcriptomic changes in bronchial epithelial cells from cystic fibrosis patients

ABSTRACT: BACKGROUND: At the moment, lung transplantation remains the best therapeutic option to ensure a better survival rate of cystic fibrosis (CF) patients, but, unfortunately, immunosuppressive therapies, often employed at high dosages to avoid acute rejection and a rapid pulmonary loss, may induce severe complications. In particular, patients treated with high dose of mammalian target of rapamycin inhibitors (mTOR-Is) may experience pulmonary fibrosis-associated adverse effects (including BOOP). Although epithelial to mesenchymal transition (EMT) of airway cells has a central role in this process, the complete biological machinery involved is not completely clarified. METHODS: In order to improve our knowledge on this process, primary bronchial epithelial cell cultures carrying F508del (BE91/3) were treated with 5 and 100 nM EVE for 24 hours. Subsequently, the total RNA extracted by these cells was hybridized to the Human HT-12 v3 Expression BeadChip (Illumina). Microarray results were validated by Real-Time PCR. Transepithelial resistance was measured by Millicell-ERS ohmmeter. RESULTS: High dosage EVE induced a significant up-regulation of 42 genes and a down-regulation of 12 genes. After pathway analysis by GSEA, we found that most of them were implicated in the inflammatory and pro-fibrotic pathway. Real Time PCR confirmed that 100 nM EVE was able to up-regulate some identified genes (IL-1a, IL-8, Pim-1 Oncogene) as well as pro-fibrotic elements (such as: a-SMA, fibronectin, Transforming Growth Factor Beta 1, connective tissue growth factor and metalloproteinase 12). Additionally, high dosage of EVE was also able to reduce the transepithelial resistance. In contrast, lower level of EVE did not produce similar effects. CONCLUSIONS: These results confirmed, also in cells derived from CF patients, the pro-fibrotic inflammatory-induced activity of high dosage EVE and revealed in these cells an interesting multi-factorial biological machinery associated with the above-mentioned process. Finally, they suggest that if employed in transplanted CF patients, EVE dosage should be reduced to the lowest possible to minimize pulmonary adverse effects.

ORGANISM(S): Homo sapiens

SUBMITTER: Fabio Sallustio 

PROVIDER: E-MTAB-8123 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress