Recombinant CFTR detection in CF tracheal epithelial cells following in vitro liposome-mediated gene transfer

Alessia Colosimo; Stefania Scarpino; Federica Sangiuolo; Sabrina Di Sario; Giuseppe Mossa; Giuseppe Novelli; Bruno Dallapiccola

Recombinant CFTR detection in CF tracheal epithelial cells following in vitro liposome-mediated gene transfer

Alessia Colosimo, Stefania Scarpino, Federica Sangiuolo, Sabrina Di Sario, Giuseppe Mossa, Giuseppe Novelli, Bruno Dallapiccola

Research output: Contribution to journal › Article › peer-review

Abstract

The efficacy of CFTR gene transfer mediated by cationic liposomes Dc-Chol/DOPE into cystic fibrosis (CF) tracheal epithelial cells carrying defective processing mutations (S549N/N1303K), was assessed by studying mRNA and protein expression of the recombinant product. Appreciable levels of mRNA transcripts were detected 48 h after transfection, while complete translocation of the recombinant CFTR to the apical membrane of epithelial cells was observed after 72 h following transfection. Our results suggest that in vitro restoration of a normal CFTR processing and migration to the cell plasmalemma requires 72 h at least as demonstrated by immunocytofluorescence using the monoclonal antibody MATG 1016. These findings are relevant onto gene transfer phase I clinical studies.

Original language	English
Pages (from-to)	723-729
Number of pages	7
Journal	Biochemistry and Molecular Biology International
Volume	42
Issue number	4
Publication status	Published - Jul 1997

Keywords

Cationic liposomes
Cystic fibrosis
Gene therapy
Immunocytochemistry
Tracheal epithelial cells

ASJC Scopus subject areas

Biochemistry
Genetics
Molecular Biology

Cite this

Recombinant CFTR detection in CF tracheal epithelial cells following in vitro liposome-mediated gene transfer. / Colosimo, Alessia; Scarpino, Stefania; Sangiuolo, Federica; Di Sario, Sabrina; Mossa, Giuseppe; Novelli, Giuseppe; Dallapiccola, Bruno.

In: Biochemistry and Molecular Biology International, Vol. 42, No. 4, 07.1997, p. 723-729.

Research output: Contribution to journal › Article › peer-review

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abstract = "The efficacy of CFTR gene transfer mediated by cationic liposomes Dc-Chol/DOPE into cystic fibrosis (CF) tracheal epithelial cells carrying defective processing mutations (S549N/N1303K), was assessed by studying mRNA and protein expression of the recombinant product. Appreciable levels of mRNA transcripts were detected 48 h after transfection, while complete translocation of the recombinant CFTR to the apical membrane of epithelial cells was observed after 72 h following transfection. Our results suggest that in vitro restoration of a normal CFTR processing and migration to the cell plasmalemma requires 72 h at least as demonstrated by immunocytofluorescence using the monoclonal antibody MATG 1016. These findings are relevant onto gene transfer phase I clinical studies.",

keywords = "Cationic liposomes, Cystic fibrosis, Gene therapy, Immunocytochemistry, Tracheal epithelial cells",

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AU - Colosimo, Alessia

AU - Scarpino, Stefania

AU - Sangiuolo, Federica

AU - Di Sario, Sabrina

AU - Mossa, Giuseppe

AU - Novelli, Giuseppe

AU - Dallapiccola, Bruno

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N2 - The efficacy of CFTR gene transfer mediated by cationic liposomes Dc-Chol/DOPE into cystic fibrosis (CF) tracheal epithelial cells carrying defective processing mutations (S549N/N1303K), was assessed by studying mRNA and protein expression of the recombinant product. Appreciable levels of mRNA transcripts were detected 48 h after transfection, while complete translocation of the recombinant CFTR to the apical membrane of epithelial cells was observed after 72 h following transfection. Our results suggest that in vitro restoration of a normal CFTR processing and migration to the cell plasmalemma requires 72 h at least as demonstrated by immunocytofluorescence using the monoclonal antibody MATG 1016. These findings are relevant onto gene transfer phase I clinical studies.

AB - The efficacy of CFTR gene transfer mediated by cationic liposomes Dc-Chol/DOPE into cystic fibrosis (CF) tracheal epithelial cells carrying defective processing mutations (S549N/N1303K), was assessed by studying mRNA and protein expression of the recombinant product. Appreciable levels of mRNA transcripts were detected 48 h after transfection, while complete translocation of the recombinant CFTR to the apical membrane of epithelial cells was observed after 72 h following transfection. Our results suggest that in vitro restoration of a normal CFTR processing and migration to the cell plasmalemma requires 72 h at least as demonstrated by immunocytofluorescence using the monoclonal antibody MATG 1016. These findings are relevant onto gene transfer phase I clinical studies.

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KW - Cystic fibrosis

KW - Gene therapy

KW - Immunocytochemistry

KW - Tracheal epithelial cells

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Recombinant CFTR detection in CF tracheal epithelial cells following in vitro liposome-mediated gene transfer

Abstract

Keywords

ASJC Scopus subject areas

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