Ability of adenovirus vectors containing different CFTR transcriptional cassettes to correct ion transport defects in CF cells

被引:25
作者
Jiang, CW
OConnor, SP
Armentano, D
Berthelette, PB
Schiavi, SC
Jefferson, DM
Smith, AE
Wadsworth, SC
Cheng, SH
机构
[1] GENZYME CORP, FRAMINGHAM, MA 01701 USA
[2] TUFTS UNIV, NEW ENGLAND MED CTR, SCH MED, BOSTON, MA 02111 USA
关键词
chloride channel; cystic fibrosis; gene transfer; cystic fibrosis transmembrane conductance regulator; sodium channel;
D O I
10.1152/ajplung.1996.271.4.L527
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Cystic fibrosis (CF) airway epithelial cells exhibit defective adenosine 3',5'-cyclic monophosphate (cAMP)-mediated chloride (Cl-) secretion, abnormal hyperabsorption of sodium (Na+), and aberrant fluid transport. Adenovirus-mediated transduction of cystic fibrosis transmembrane conductance regulator (CFTR) corrects these ion and fluid transport abnormalities in CF cells. However, several challenges remain pertaining to the use of adenovirus vectors for gene delivery, including the efficiency of gene transfer and the host response to the vector. To improve the efficacy of adenovirus-mediated gene transfer, we have constructed a series of recombinant adenoviruses containing different CFTR transcriptional units, and we have evaluated their relative ability to correct electrolyte and fluid transport in polarized CF airway epithelial cells. The ability of the vectors to correct the CF Cl- transport defects was greatest when the human cytomegalovirus promoter was used. The E1a and phosphoglycerate kinase promoters resulted in the greatest persistence of functional CFTR expression. Efficacy of gene expression by recombinant adenoviruses improved as the cells were treated with increasing multiplicities of infection, as the duration of viral contact with the target cells was lengthened, and when the virus concentration was increased. Transduction of functional CFTR Cl- channel activity reversed the abnormal Na+ hyperabsorption observed in CF cells in a dose-dependent manner, suggesting that Na+ channel activity is downregulated by CFTR. Although efficient correction of both cAMP-mediated Cl- transport and fluid secretion could be achieved readily with these vectors, normalization of the Na+ absorption required vector administration at high multiplicities of infection.
引用
收藏
页码:L527 / L537
页数:11
相关论文
共 47 条
[1]  
APPERLEY JF, 1991, BLOOD, V78, P310
[2]   CHARACTERIZATION OF AN ADENOVIRUS GENE-TRANSFER VECTOR CONTAINING AN E4 DELETION [J].
ARMENTANO, D ;
SOOKDEO, CC ;
HEHIR, KM ;
GREGORY, RJ ;
STGEORGE, JA ;
PRINCE, GA ;
WADSWORTH, SC ;
SMITH, AE .
HUMAN GENE THERAPY, 1995, 6 (10) :1343-1353
[3]  
Boat T, 1989, CYSTIC FIBROSIS META, P2649
[4]   A VERY STRONG ENHANCER IS LOCATED UPSTREAM OF AN IMMEDIATE EARLY GENE OF HUMAN CYTOMEGALO-VIRUS [J].
BOSHART, M ;
WEBER, F ;
JAHN, G ;
DORSCHHASLER, K ;
FLECKENSTEIN, B ;
SCHAFFNER, W .
CELL, 1985, 41 (02) :521-530
[5]   NA+ TRANSPORT IN CYSTIC-FIBROSIS RESPIRATORY EPITHELIA - ABNORMAL BASAL RATE AND RESPONSE TO ADENYLATE-CYCLASE ACTIVATION [J].
BOUCHER, RC ;
STUTTS, MJ ;
KNOWLES, MR ;
CANTLEY, L ;
GATZY, JT .
JOURNAL OF CLINICAL INVESTIGATION, 1986, 78 (05) :1245-1252
[6]   EVIDENCE FOR REDUCED CL- AND INCREASED NA+ PERMEABILITY IN CYSTIC-FIBROSIS HUMAN PRIMARY-CELL CULTURES [J].
BOUCHER, RC ;
COTTON, CU ;
GATZY, JT ;
KNOWLES, MR ;
YANKASKAS, JR .
JOURNAL OF PHYSIOLOGY-LONDON, 1988, 405 :77-103
[7]   ACUTE RESPONSES OF NONHUMAN-PRIMATES TO AIRWAY DELIVERY OF AN ADENOVIRUS VECTOR CONTAINING THE HUMAN CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR CDNA [J].
BRODY, SL ;
METZGER, M ;
DANEL, C ;
ROSENFELD, MA ;
CRYSTAL, RG .
HUMAN GENE THERAPY, 1994, 5 (07) :821-836
[8]   INVIVO PROMOTER ACTIVITY AND TRANSGENE EXPRESSION IN MAMMALIAN SOMATIC TISSUES EVALUATED BY USING PARTICLE BOMBARDMENT [J].
CHENG, L ;
ZIEGELHOFFER, PR ;
YANG, NS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (10) :4455-4459
[9]   ADMINISTRATION OF AN ADENOVIRUS CONTAINING THE HUMAN CFTR CDNA TO THE RESPIRATORY-TRACT OF INDIVIDUALS WITH CYSTIC-FIBROSIS [J].
CRYSTAL, RG ;
MCELVANEY, NG ;
ROSENFELD, MA ;
CHU, CS ;
MASTRANGELI, A ;
HAY, JG ;
BRODY, SL ;
JAFFE, HA ;
EISSA, NT ;
DANEL, C .
NATURE GENETICS, 1994, 8 (01) :42-51
[10]  
DORIN JR, 1994, CYSTIC FIBROSIS CURR, P3