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

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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",

author = "Alessia Colosimo and Stefania Scarpino and Federica Sangiuolo and {Di Sario}, Sabrina and Giuseppe Mossa and Giuseppe Novelli and Bruno Dallapiccola",

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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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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 - 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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