Generation of expression plasmids for angiostatin, endostatin and TIMP-2 for cancer gene therapy

Stefano Indraccolo, S. Minuzzo, E. Gola, W. Habeler, F. Carrozzino, D. Noonan, A. Albini, L. Santi, A. Amadori, L. Chieco-Bianchi

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)251-256
Number of pages6
JournalInternational Journal of Biological Markers
Volume14
Issue number4
Publication statusPublished - Oct 1999

Keywords

  • Angiostatin
  • Endostatin
  • Gene therapy
  • Retroviral vectors
  • TIMP-2

ASJC Scopus subject areas

  • Immunology
  • Biochemistry

Fingerprint Dive into the research topics of 'Generation of expression plasmids for angiostatin, endostatin and TIMP-2 for cancer gene therapy'. Together they form a unique fingerprint.

Cite this