A Peptide-Nucleic Acid Targeting miR-335-5p Enhances Expression of Cystic Fibrosis Transmembrane Conductance Regulator (<em>CFTR</em>) Gene with the Possible Involvement of the CFTR Scaffolding Protein NHERF1
Anna Tamanini,
Enrica Fabbri,
Tiziana Jakova,
Jessica Gasparello,
Alex Manicardi,
Roberto Corradini,
Alessia Finotti,
Monica Borgatti,
Ilaria Lampronti,
Silvia Munari,
Maria Cristina Dechecchi,
Giulio Cabrini,
Roberto Gambari
Affiliations
Anna Tamanini
Section of Molecular Pathology, Department of Pathology and Diagnostics, University-Hospital of Verona, 37126 Verona, Italy
Enrica Fabbri
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy
Tiziana Jakova
Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, 43124 Parma, Italy
Jessica Gasparello
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy
Alex Manicardi
Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, 43124 Parma, Italy
Roberto Corradini
Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, 43124 Parma, Italy
Alessia Finotti
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy
Monica Borgatti
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy
Ilaria Lampronti
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy
Silvia Munari
Section of Molecular Pathology, Department of Pathology and Diagnostics, University-Hospital of Verona, 37126 Verona, Italy
Maria Cristina Dechecchi
Department of Neurosciences, Biomedicine and Movement, University of Verona, 37100 Verona, Italy
Giulio Cabrini
Research Center on Innovative Therapies for Cystic Fibrosis, University of Ferrara, 44121 Ferrara, Italy
Roberto Gambari
Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy
(1) Background: Up-regulation of the Cystic Fibrosis Transmembrane Conductance Regulator gene (CFTR) might be of great relevance for the development of therapeutic protocols for cystic fibrosis (CF). MicroRNAs are deeply involved in the regulation of CFTR and scaffolding proteins (such as NHERF1, NHERF2 and Ezrin). (2) Methods: Content of miRNAs and mRNAs was analyzed by RT-qPCR, while the CFTR and NHERF1 production was analyzed by Western blotting. (3) Results: The results here described show that the CFTR scaffolding protein NHERF1 can be up-regulated in bronchial epithelial Calu-3 cells by a peptide-nucleic acid (PNA) targeting miR-335-5p, predicted to bind to the 3′-UTR sequence of the NHERF1 mRNA. Treatment of Calu-3 cells with this PNA (R8-PNA-a335) causes also up-regulation of CFTR. (4) Conclusions: We propose miR-335-5p targeting as a strategy to increase CFTR. While the efficiency of PNA-based targeting of miR-335-5p should be verified as a therapeutic strategy in CF caused by stop-codon mutation of the CFTR gene, this approach might give appreciable results in CF cells carrying other mutations impairing the processing or stability of CFTR protein, supporting its application in personalized therapy for precision medicine.
