Dual therapeutic utility of proteasome modulating agents for pharmaco-gene therapy of the cystic fibrosis airway.
Pharmacologic- and gene-based therapies have historically been developed as two independent therapeutic platforms for cystic fibrosis (CF) lung disease. Inhibition of the dysregulated epithelial Na channel (ENaC) is one pharmacologic approach to enhance airway clearance in CF. We investigated pharmacologic approaches to enhance CFTR gene delivery with recombinant adeno-associated virus (rAAV) and identified compounds that significantly improved viral transduction while simultaneously inhibiting ENaC activity through an unrelated mechanism. Treatment of human CF airway epithelia with proteasome modulating agents (LLnL and doxorubicin) at the time of rAAV2 or rAAV2/5 infection dramatically enhanced CFTR gene delivery and correction of CFTR-mediated short-circuit currents. Surprisingly, these agents also facilitated long-term (15-day) functional inhibition of ENaC currents independent of CFTR vector administration. Inhibition of ENaC activity was predominantly attributed to a doxorubicin-dependent decrease in gamma-ENaC subunit mRNA expression and an increase in gamma-ENaC promoter methylation. This is the first report to describe the identification of compounds with dual therapeutic action that are able to enhance the efficacy of CFTR gene therapy to the airway while simultaneously ameliorating primary aspects of CF disease pathophysiology. The identification of such compounds mark a new area for drug development, not only for CF, but also for other gene therapy disease targets.
Zhang
Liang N
LN
Department of Anatomy and Cell Biology, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
Karp
Phil
P
Gerard
Christopher J
CJ
Pastor
Eric
E
Laux
Douglas
D
Munson
Keith
K
Yan
Ziying
Z
Liu
Xiaoming
X
Godwin
Simon
S
Thomas
Christie P
CP
Zabner
Joseph
J
Shi
Huidong
H
Caldwell
Charles W
CW
Peluso
Richard
R
Carter
Barrie
B
Engelhardt
John F
JF
D000584:Amiloride / Q000494:pharmacology; D016764:Cell Polarity; D002478:Cells, Cultured; D018899:CpG Islands / Q000235:genetics; D003550:Cystic Fibrosis / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D019175:DNA Methylation; D000229:Dependovirus / Q000235:genetics; D004317:Doxorubicin / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D053503:Epithelial Sodium Channels; D015316:Genetic Therapy / Q000379:methods*; D016679:Genome, Viral; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism; D013997:Time Factors
C505032:CFTR protein, human; D053503:Epithelial Sodium Channels; D061988:Proteasome Inhibitors; D012333:RNA, Messenger; D015222:Sodium Channels; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000584:Amiloride; D004317:Doxorubicin; D012964:Sodium; D046988:Proteasome Endopeptidase Complex