Transfer of a constitutive viral pormoter-cystic fibrosis transmembrance conductance regulator cDNA to human epithelial cells conveys resistance to down-regulation of cAMP-regulated Cl- secretion in the presence of inflammatory stimuli

Nobuyuki Kobayashi, Eugene R. Rosenthal, Kunihiko Yoshimura, Ronald Crystal

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene can be downregulated by inflammatory stimuli such as phorbol myristate acetate (PMA). Since the respiratory manifestations of cystic fibrosis (CF) are characterized by intense chronic airway inflammation very early in life, successful gene therapy for CF will require that expression of the transferred normal CFTR gene be resistant to down-regulation by inflammatory mediators. To evaluate the concept that a viral promoter- human CFTR cDNA unit would be resistant to this form of down-regulation, a retrovirus promoter (5′ long terminal repeat of the Moloney murine leukemia virus) - human CFTR cDNA unit was transferred to T84 human colon carcinoma cell line using a retrovirus vector. Exposure of the retrovirus-modified T84 cells to PMA resulted in down-regulation of the endogenous CFTR mRNA transcripts (6.5 kb), but did not affect the level of exogenous CFTR transcripts (8.0 kb). Importantly, in parallel with the persistence of the exogenous CFTR transcripts, the modified cells still maintained cAMP-regulated Cl- secretion in the presence of PMA. These in vitro data suggest that a constitutive viral promoter -CFTR cDNA unit should be resistant to modulation by inflammatory stimuli, a likely requirement for successful gene therapy for CF.

Original languageEnglish
Pages (from-to)4470-4476
Number of pages7
JournalNucleic Acids Research
Volume22
Issue number21
DOIs
Publication statusPublished - 25 Oct 1994
Externally publishedYes

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Cystic Fibrosis
Down-Regulation
Complementary DNA
Epithelial Cells
Tetradecanoylphorbol Acetate
Retroviridae
Regulator Genes
Genetic Therapy
Moloney murine leukemia virus
Terminal Repeat Sequences
Colon
Inflammation
Carcinoma
Cell Line
Messenger RNA

ASJC Scopus subject areas

  • Genetics

Cite this

@article{90d1b1ad19f84154a4557c892a067027,
title = "Transfer of a constitutive viral pormoter-cystic fibrosis transmembrance conductance regulator cDNA to human epithelial cells conveys resistance to down-regulation of cAMP-regulated Cl- secretion in the presence of inflammatory stimuli",
abstract = "The expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene can be downregulated by inflammatory stimuli such as phorbol myristate acetate (PMA). Since the respiratory manifestations of cystic fibrosis (CF) are characterized by intense chronic airway inflammation very early in life, successful gene therapy for CF will require that expression of the transferred normal CFTR gene be resistant to down-regulation by inflammatory mediators. To evaluate the concept that a viral promoter- human CFTR cDNA unit would be resistant to this form of down-regulation, a retrovirus promoter (5′ long terminal repeat of the Moloney murine leukemia virus) - human CFTR cDNA unit was transferred to T84 human colon carcinoma cell line using a retrovirus vector. Exposure of the retrovirus-modified T84 cells to PMA resulted in down-regulation of the endogenous CFTR mRNA transcripts (6.5 kb), but did not affect the level of exogenous CFTR transcripts (8.0 kb). Importantly, in parallel with the persistence of the exogenous CFTR transcripts, the modified cells still maintained cAMP-regulated Cl- secretion in the presence of PMA. These in vitro data suggest that a constitutive viral promoter -CFTR cDNA unit should be resistant to modulation by inflammatory stimuli, a likely requirement for successful gene therapy for CF.",
author = "Nobuyuki Kobayashi and Rosenthal, {Eugene R.} and Kunihiko Yoshimura and Ronald Crystal",
year = "1994",
month = "10",
day = "25",
doi = "10.1093/nar/22.21.4470",
language = "English",
volume = "22",
pages = "4470--4476",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "21",

}

TY - JOUR

T1 - Transfer of a constitutive viral pormoter-cystic fibrosis transmembrance conductance regulator cDNA to human epithelial cells conveys resistance to down-regulation of cAMP-regulated Cl- secretion in the presence of inflammatory stimuli

AU - Kobayashi, Nobuyuki

AU - Rosenthal, Eugene R.

AU - Yoshimura, Kunihiko

AU - Crystal, Ronald

PY - 1994/10/25

Y1 - 1994/10/25

N2 - The expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene can be downregulated by inflammatory stimuli such as phorbol myristate acetate (PMA). Since the respiratory manifestations of cystic fibrosis (CF) are characterized by intense chronic airway inflammation very early in life, successful gene therapy for CF will require that expression of the transferred normal CFTR gene be resistant to down-regulation by inflammatory mediators. To evaluate the concept that a viral promoter- human CFTR cDNA unit would be resistant to this form of down-regulation, a retrovirus promoter (5′ long terminal repeat of the Moloney murine leukemia virus) - human CFTR cDNA unit was transferred to T84 human colon carcinoma cell line using a retrovirus vector. Exposure of the retrovirus-modified T84 cells to PMA resulted in down-regulation of the endogenous CFTR mRNA transcripts (6.5 kb), but did not affect the level of exogenous CFTR transcripts (8.0 kb). Importantly, in parallel with the persistence of the exogenous CFTR transcripts, the modified cells still maintained cAMP-regulated Cl- secretion in the presence of PMA. These in vitro data suggest that a constitutive viral promoter -CFTR cDNA unit should be resistant to modulation by inflammatory stimuli, a likely requirement for successful gene therapy for CF.

AB - The expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene can be downregulated by inflammatory stimuli such as phorbol myristate acetate (PMA). Since the respiratory manifestations of cystic fibrosis (CF) are characterized by intense chronic airway inflammation very early in life, successful gene therapy for CF will require that expression of the transferred normal CFTR gene be resistant to down-regulation by inflammatory mediators. To evaluate the concept that a viral promoter- human CFTR cDNA unit would be resistant to this form of down-regulation, a retrovirus promoter (5′ long terminal repeat of the Moloney murine leukemia virus) - human CFTR cDNA unit was transferred to T84 human colon carcinoma cell line using a retrovirus vector. Exposure of the retrovirus-modified T84 cells to PMA resulted in down-regulation of the endogenous CFTR mRNA transcripts (6.5 kb), but did not affect the level of exogenous CFTR transcripts (8.0 kb). Importantly, in parallel with the persistence of the exogenous CFTR transcripts, the modified cells still maintained cAMP-regulated Cl- secretion in the presence of PMA. These in vitro data suggest that a constitutive viral promoter -CFTR cDNA unit should be resistant to modulation by inflammatory stimuli, a likely requirement for successful gene therapy for CF.

UR - http://www.scopus.com/inward/record.url?scp=0028072786&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028072786&partnerID=8YFLogxK

U2 - 10.1093/nar/22.21.4470

DO - 10.1093/nar/22.21.4470

M3 - Article

C2 - 7526342

AN - SCOPUS:0028072786

VL - 22

SP - 4470

EP - 4476

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 21

ER -