Cystic Fibrosis: Nasal epithelial cells: a novel ex vivo model to assess the role of novel mutations and the effect of drugs

Il 30/01/2015 ore 14.30 - 15.30

Sala Conferenze, CNR Area della Ricerca NA1, Via P. Castellino, 111 80131 Napoli

Prof Giuseppe Castaldo (CEINGE-Biotecnologie avanzate, Napoli and Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II) will report recent results gained by his studies on Cystic Fibrosis (CF). CF patients have specific symptoms, increased sweat chloride and 2 mutations in the CFTR gene, even if first level molecular analysis lacks to identify mutations in about 15% of alleles. In addition to the classic CF, atypical forms (CFTR-RD) have been described that do not meet diagnostic criteria for CF, show normal or borderline sweat test and often a single mutation. Sequencing analysis can identify a second mutation but in several cases it may be a mutation of uncertain pathogenic significance requiring a complex in vitro analysis to determine its pathogenic effect. Furthermore, a strongly heterogeneous clinical expression was reported in CF patients bearing the same genotype and in pairs of affected siblings, suggesting that modifier factors of CF phenotype may have a role.

In the last years, we identified mutations within the non-coding regions of CFTR, including the large promoter area at the 5’ of the gene, or the 3’UTR (that is the target of miRNA inhibitory effect). Such mutations may impair CFTR gene expression with a different effect on different cell lines thus, putatively acting as modifier factors of CF clinical expression in different tissues and organs. Finally, drugs are now available that may correct or potentiate the CFTR protein activity in CF patients bearing specific mutations.

The model of epithelial nasal cells from patients allows to study the effect of mutations directly in an ex vivo system obtained from patients. Prof Castaldo's group developed this model and a series of advanced molecular technologies that may help to define the molecular effect of CFTR mutations of uncertain significance (i.e., monoclonal antibodies to define altered CFTR topogenesis; RT-PCR-electrophoresis to assess the splicing effect of mutations that lie in exon-intron boundaries; halide-sensitive fluorescent assay to quantitate the CFTR gating activity).

The same model helps to define the effect of mutations in regulatory regions of CFTR gene (i.e., quantitative RT-PCR analysis to verify if the mutation would cause a reduced gene expression). Finally, the model contributes to evaluate the effect of novel drugs that potentiate or correct the CFTR protein due to effect of specific mutations before to use the drug on patients.    

Organizzato da:
IBP, Cnr

Referente organizzativo:
Alberto Luini
CNR - Istituto di biochimica delle proteine
Via P. Castellino, 111 80131 Napoli
a.luini@ibp.cnr.it
081/6132535

Modalità di accesso: ingresso libero