Abstract
Resolution of alveolar edema depends on active ion transport by sodium pumps located on the basolateral surface of alveolar epithelial cells (AECs), suggesting that upregulation of sodium pump activity may facilitate clearance of edema fluid. We have investigated the use of lentiviral vectors to augment sodium pump activity via gene transfer of sodium pump subunits to AECs. Full-length cDNA for the α1 or β1 subunit of rat Na+,K+-ATPase was cloned into the lentiviral vector pRRLsin.hCMV.IRES.EGFP. Rat AECs in primary culture were transduced on day 4 with lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein G. Transduction with lentiviral vectors encoding either α1 subunit (Lenti-α1-EGFP) or β1 subunit (Lenti-β1-EGFP) led to dose-dependent increases in mRNA and protein for the corresponding subunit. Transduction with Lenti- β1-EGFP was accompanied by coordinate upregulation of endogenous α1 expression, whereas endogenous β1 expression was unchanged after transduction with Lenti-α1-EGFP. Consistent with these findings, transduction with Lenti-β1- EGFP, but not Lenti-α1-EGFP, led to augmentation of sodium pump activity as a result of increases in Na+,K+-ATPase holoenzyme. Sodium pump α2 subunit and sodium channel protein did not change after Lenti-β1-EGFP transduction. These results demonstrate that overall sodium pump activity can be efficiently upregulated in AECs specifically via gene transfer of the sodium pump β1 subunit and support the feasibility of lentivirus-mediated gene transfer to augment alveolar fluid clearance.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 457-468 |
| Number of pages | 12 |
| Journal | Human Gene Therapy |
| Volume | 15 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 2004 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Genetics
Cite this
Subunit-specific coordinate upregulation of sodium pump activity in alveolar epithelial cells by lentivirus-mediated gene transfer. / Qiao, Renli; Zhou, Beiyun; Harboe-Schmidt, Erik; Kasahara, Noriyuki; Kim, Kwang Jin; Liebler, Janice M.; Crandall, Edward D.; Borok, Zea.
In: Human Gene Therapy, Vol. 15, No. 5, 05.2004, p. 457-468.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Subunit-specific coordinate upregulation of sodium pump activity in alveolar epithelial cells by lentivirus-mediated gene transfer
AU - Qiao, Renli
AU - Zhou, Beiyun
AU - Harboe-Schmidt, Erik
AU - Kasahara, Noriyuki
AU - Kim, Kwang Jin
AU - Liebler, Janice M.
AU - Crandall, Edward D.
AU - Borok, Zea
PY - 2004/5
Y1 - 2004/5
N2 - Resolution of alveolar edema depends on active ion transport by sodium pumps located on the basolateral surface of alveolar epithelial cells (AECs), suggesting that upregulation of sodium pump activity may facilitate clearance of edema fluid. We have investigated the use of lentiviral vectors to augment sodium pump activity via gene transfer of sodium pump subunits to AECs. Full-length cDNA for the α1 or β1 subunit of rat Na+,K+-ATPase was cloned into the lentiviral vector pRRLsin.hCMV.IRES.EGFP. Rat AECs in primary culture were transduced on day 4 with lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein G. Transduction with lentiviral vectors encoding either α1 subunit (Lenti-α1-EGFP) or β1 subunit (Lenti-β1-EGFP) led to dose-dependent increases in mRNA and protein for the corresponding subunit. Transduction with Lenti- β1-EGFP was accompanied by coordinate upregulation of endogenous α1 expression, whereas endogenous β1 expression was unchanged after transduction with Lenti-α1-EGFP. Consistent with these findings, transduction with Lenti-β1- EGFP, but not Lenti-α1-EGFP, led to augmentation of sodium pump activity as a result of increases in Na+,K+-ATPase holoenzyme. Sodium pump α2 subunit and sodium channel protein did not change after Lenti-β1-EGFP transduction. These results demonstrate that overall sodium pump activity can be efficiently upregulated in AECs specifically via gene transfer of the sodium pump β1 subunit and support the feasibility of lentivirus-mediated gene transfer to augment alveolar fluid clearance.
AB - Resolution of alveolar edema depends on active ion transport by sodium pumps located on the basolateral surface of alveolar epithelial cells (AECs), suggesting that upregulation of sodium pump activity may facilitate clearance of edema fluid. We have investigated the use of lentiviral vectors to augment sodium pump activity via gene transfer of sodium pump subunits to AECs. Full-length cDNA for the α1 or β1 subunit of rat Na+,K+-ATPase was cloned into the lentiviral vector pRRLsin.hCMV.IRES.EGFP. Rat AECs in primary culture were transduced on day 4 with lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein G. Transduction with lentiviral vectors encoding either α1 subunit (Lenti-α1-EGFP) or β1 subunit (Lenti-β1-EGFP) led to dose-dependent increases in mRNA and protein for the corresponding subunit. Transduction with Lenti- β1-EGFP was accompanied by coordinate upregulation of endogenous α1 expression, whereas endogenous β1 expression was unchanged after transduction with Lenti-α1-EGFP. Consistent with these findings, transduction with Lenti-β1- EGFP, but not Lenti-α1-EGFP, led to augmentation of sodium pump activity as a result of increases in Na+,K+-ATPase holoenzyme. Sodium pump α2 subunit and sodium channel protein did not change after Lenti-β1-EGFP transduction. These results demonstrate that overall sodium pump activity can be efficiently upregulated in AECs specifically via gene transfer of the sodium pump β1 subunit and support the feasibility of lentivirus-mediated gene transfer to augment alveolar fluid clearance.
UR - http://www.scopus.com/inward/record.url?scp=2442567911&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=2442567911&partnerID=8YFLogxK
U2 - 10.1089/10430340460745784
DO - 10.1089/10430340460745784
M3 - Article
C2 - 15144576
AN - SCOPUS:2442567911
VL - 15
SP - 457
EP - 468
JO - Human Gene Therapy
JF - Human Gene Therapy
SN - 1043-0342
IS - 5
ER -