Abstract
RNA interference, due to its target specificity, may be highly effective as a novel therapeutic modality, but direct delivery of synthetic small interfering RNA still remains a major obstacle for this approach. To induce long-term expression and specific gene silencing, novel delivery vector system is also required. In this study, we have generated an efficient oncolytic adenovirus (Ad)-based short hairpin (shRNA) expression system (Ad-ΔB7-U6shIL8) against IL-8, a potent proangiogenic factor. To demonstrate IL-8-specificity of this newly engineered Ad-based shRNA, we also manufactured replication-incompetent Ads (Ad-ΔE1-CMVshIL8 and Ad-ΔE1-U6shIL8) under the control of the cytomegalovirus (CMV) and U6 promoters, respectively. Ad-ΔE1-U6shIL8 was highly effective in reducing IL-8 expression, and was much more effective in driving IL-8-specific shRNA than the CMV promoter-driven vector. The reduced IL-8 expression then translated into decreased angiogenesis in vitro as measured by migration, tube formation and rat aortic ring sprouting assays. In addition to its effect on endothelial cells, Ad-ΔE1-U6shIL8 also effectively suppressed the migration and invasion of cancer cells. In vivo, intratumoral injection of Ad-ΔB7-U6shIL8 significantly inhibited the growth of Hep3B and A549 human tumor xenografts. Histopathological analysis of Ad-ΔB7-U6shIL8-treated tumors revealed an increase in apoptotic cells and a reduction in vessel density. Finally, Ad-ΔB7-U6shIL8 was also shown to inhibit the growth of disseminated MDA-MB-231 breast cancer metastases. Taken together, these findings demonstrate the utility and antitumor effectiveness of oncolytic Ad expressing shRNA against IL-8.
| Original language | English |
|---|---|
| Pages (from-to) | 635-651 |
| Number of pages | 17 |
| Journal | Gene Therapy |
| Volume | 15 |
| Issue number | 9 |
| DOIs | |
| State | Published - 2008 May 1 |
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Short hairpin RNA-expressing oncolytic adenovirus-mediated inhibition of IL-8 : Effects on antiangiogenesis and tumor growth inhibition. / Yoo, J. Y.; Kim, J. H.; Kim, J.; Huang, J. H.; Zhang, S. N.; Kang, Y. A.; Kim, H.; Yun, Chae Ok.
In: Gene Therapy, Vol. 15, No. 9, 01.05.2008, p. 635-651.Research output: Contribution to journal › Article
TY - JOUR
T1 - Short hairpin RNA-expressing oncolytic adenovirus-mediated inhibition of IL-8
T2 - Effects on antiangiogenesis and tumor growth inhibition
AU - Yoo, J. Y.
AU - Kim, J. H.
AU - Kim, J.
AU - Huang, J. H.
AU - Zhang, S. N.
AU - Kang, Y. A.
AU - Kim, H.
AU - Yun, Chae Ok
PY - 2008/5/1
Y1 - 2008/5/1
N2 - RNA interference, due to its target specificity, may be highly effective as a novel therapeutic modality, but direct delivery of synthetic small interfering RNA still remains a major obstacle for this approach. To induce long-term expression and specific gene silencing, novel delivery vector system is also required. In this study, we have generated an efficient oncolytic adenovirus (Ad)-based short hairpin (shRNA) expression system (Ad-ΔB7-U6shIL8) against IL-8, a potent proangiogenic factor. To demonstrate IL-8-specificity of this newly engineered Ad-based shRNA, we also manufactured replication-incompetent Ads (Ad-ΔE1-CMVshIL8 and Ad-ΔE1-U6shIL8) under the control of the cytomegalovirus (CMV) and U6 promoters, respectively. Ad-ΔE1-U6shIL8 was highly effective in reducing IL-8 expression, and was much more effective in driving IL-8-specific shRNA than the CMV promoter-driven vector. The reduced IL-8 expression then translated into decreased angiogenesis in vitro as measured by migration, tube formation and rat aortic ring sprouting assays. In addition to its effect on endothelial cells, Ad-ΔE1-U6shIL8 also effectively suppressed the migration and invasion of cancer cells. In vivo, intratumoral injection of Ad-ΔB7-U6shIL8 significantly inhibited the growth of Hep3B and A549 human tumor xenografts. Histopathological analysis of Ad-ΔB7-U6shIL8-treated tumors revealed an increase in apoptotic cells and a reduction in vessel density. Finally, Ad-ΔB7-U6shIL8 was also shown to inhibit the growth of disseminated MDA-MB-231 breast cancer metastases. Taken together, these findings demonstrate the utility and antitumor effectiveness of oncolytic Ad expressing shRNA against IL-8.
AB - RNA interference, due to its target specificity, may be highly effective as a novel therapeutic modality, but direct delivery of synthetic small interfering RNA still remains a major obstacle for this approach. To induce long-term expression and specific gene silencing, novel delivery vector system is also required. In this study, we have generated an efficient oncolytic adenovirus (Ad)-based short hairpin (shRNA) expression system (Ad-ΔB7-U6shIL8) against IL-8, a potent proangiogenic factor. To demonstrate IL-8-specificity of this newly engineered Ad-based shRNA, we also manufactured replication-incompetent Ads (Ad-ΔE1-CMVshIL8 and Ad-ΔE1-U6shIL8) under the control of the cytomegalovirus (CMV) and U6 promoters, respectively. Ad-ΔE1-U6shIL8 was highly effective in reducing IL-8 expression, and was much more effective in driving IL-8-specific shRNA than the CMV promoter-driven vector. The reduced IL-8 expression then translated into decreased angiogenesis in vitro as measured by migration, tube formation and rat aortic ring sprouting assays. In addition to its effect on endothelial cells, Ad-ΔE1-U6shIL8 also effectively suppressed the migration and invasion of cancer cells. In vivo, intratumoral injection of Ad-ΔB7-U6shIL8 significantly inhibited the growth of Hep3B and A549 human tumor xenografts. Histopathological analysis of Ad-ΔB7-U6shIL8-treated tumors revealed an increase in apoptotic cells and a reduction in vessel density. Finally, Ad-ΔB7-U6shIL8 was also shown to inhibit the growth of disseminated MDA-MB-231 breast cancer metastases. Taken together, these findings demonstrate the utility and antitumor effectiveness of oncolytic Ad expressing shRNA against IL-8.
UR - http://www.scopus.com/inward/record.url?scp=42149120089&partnerID=8YFLogxK
U2 - 10.1038/gt.2008.3
DO - 10.1038/gt.2008.3
M3 - Article
C2 - 18273054
AN - SCOPUS:42149120089
VL - 15
SP - 635
EP - 651
JO - Gene Therapy
JF - Gene Therapy
SN - 0969-7128
IS - 9
ER -