Abstract
Antiangiogenic therapy may represent a promising approach to cancer treatment. Indeed, the efficacy of endogenous angiogenesis inhibitors, including angiostatin, endostatin and TIMPs, has been demonstrated in many types of solid tumors in animal models. In view of the possible problems associated with long-term administration of inhibitors as recombinant proteins, we propose their delivery as nucleic acids through a gene therapy approach. To this end, eukaryotic expression constructs for murine angiostatin and endostatin as well as human TIMP-2 were generated and characterized in vitro. All constructs carry the relevant cDNAs under the control of the strong HCMV promoter/enhancer and cleavable leader signals to allow protein secretion. Expression of the angiogenesis inhibitors was detected by in vitro transcription/translation experiments as well as transfection of 293T cells, followed by Western blotting (WB) or radioimmunoprecipitation analysis of both cell lysates and supernatants (SNs). These constructs might be used for in vivo intramuscular delivery of plasmid DNA and as a set of reagents for the development of retroviral as well as adeno-associated viral (AAV) vectors expressing angiogenesis inhibitors.
| Original language | English |
|---|---|
| Pages (from-to) | 251-256 |
| Number of pages | 6 |
| Journal | International Journal of Biological Markers |
| Volume | 14 |
| Issue number | 4 |
| Publication status | Published - Oct 1999 |
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Keywords
- Angiostatin
- Endostatin
- Gene therapy
- Retroviral vectors
- TIMP-2
ASJC Scopus subject areas
- Immunology
- Biochemistry
Cite this
Generation of expression plasmids for angiostatin, endostatin and TIMP-2 for cancer gene therapy. / Indraccolo, Stefano; Minuzzo, S.; Gola, E.; Habeler, W.; Carrozzino, F.; Noonan, D.; Albini, A.; Santi, L.; Amadori, A.; Chieco-Bianchi, L.
In: International Journal of Biological Markers, Vol. 14, No. 4, 10.1999, p. 251-256.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Generation of expression plasmids for angiostatin, endostatin and TIMP-2 for cancer gene therapy
AU - Indraccolo, Stefano
AU - Minuzzo, S.
AU - Gola, E.
AU - Habeler, W.
AU - Carrozzino, F.
AU - Noonan, D.
AU - Albini, A.
AU - Santi, L.
AU - Amadori, A.
AU - Chieco-Bianchi, L.
PY - 1999/10
Y1 - 1999/10
N2 - Antiangiogenic therapy may represent a promising approach to cancer treatment. Indeed, the efficacy of endogenous angiogenesis inhibitors, including angiostatin, endostatin and TIMPs, has been demonstrated in many types of solid tumors in animal models. In view of the possible problems associated with long-term administration of inhibitors as recombinant proteins, we propose their delivery as nucleic acids through a gene therapy approach. To this end, eukaryotic expression constructs for murine angiostatin and endostatin as well as human TIMP-2 were generated and characterized in vitro. All constructs carry the relevant cDNAs under the control of the strong HCMV promoter/enhancer and cleavable leader signals to allow protein secretion. Expression of the angiogenesis inhibitors was detected by in vitro transcription/translation experiments as well as transfection of 293T cells, followed by Western blotting (WB) or radioimmunoprecipitation analysis of both cell lysates and supernatants (SNs). These constructs might be used for in vivo intramuscular delivery of plasmid DNA and as a set of reagents for the development of retroviral as well as adeno-associated viral (AAV) vectors expressing angiogenesis inhibitors.
AB - Antiangiogenic therapy may represent a promising approach to cancer treatment. Indeed, the efficacy of endogenous angiogenesis inhibitors, including angiostatin, endostatin and TIMPs, has been demonstrated in many types of solid tumors in animal models. In view of the possible problems associated with long-term administration of inhibitors as recombinant proteins, we propose their delivery as nucleic acids through a gene therapy approach. To this end, eukaryotic expression constructs for murine angiostatin and endostatin as well as human TIMP-2 were generated and characterized in vitro. All constructs carry the relevant cDNAs under the control of the strong HCMV promoter/enhancer and cleavable leader signals to allow protein secretion. Expression of the angiogenesis inhibitors was detected by in vitro transcription/translation experiments as well as transfection of 293T cells, followed by Western blotting (WB) or radioimmunoprecipitation analysis of both cell lysates and supernatants (SNs). These constructs might be used for in vivo intramuscular delivery of plasmid DNA and as a set of reagents for the development of retroviral as well as adeno-associated viral (AAV) vectors expressing angiogenesis inhibitors.
KW - Angiostatin
KW - Endostatin
KW - Gene therapy
KW - Retroviral vectors
KW - TIMP-2
UR - http://www.scopus.com/inward/record.url?scp=0033374458&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033374458&partnerID=8YFLogxK
M3 - Article
C2 - 10669955
AN - SCOPUS:0033374458
VL - 14
SP - 251
EP - 256
JO - International Journal of Biological Markers
JF - International Journal of Biological Markers
SN - 0393-6155
IS - 4
ER -