Cationic lipid and polymer-based gene delivery to human pacreatic islets

Ram I. Mahato, James Henry, Ajit S. Narang, Omaima Sabek, Daniel Fraga, Malak Kotb, Omma Gaber

Research output: Contribution to journalArticle

67 Scopus citations

Abstract

Transplantation of pancreatic islets has great potential for treating Type I diabetes. Ex vivo gene therapy may promote re-vascularization or inhibit apoptosis of the islets and promote graft. In this study, we investigated the feasibility of non-viral gene delivery using Enhanced Green Fluorescent Protein (EGFP) and human Vascular Endothelial Growth Factor (hVEGF165) expression plasmids as model reporter and therapeutic genes. LipofectAMINE/pDNA and Superfect/pDNA complexes showed high transfection efficiency in rapidly dividing Jurkat cells, but low transfection in non-dividing human islets. LipofectAMINE/pCAGGS-hVEGF transfected islets showed relatively higher levels of hVEGF than in those transfected with LipofectAMINE/pCMS-EGFP complexes or 5% glucose. To exclude endogenously secreted hVEGF, real time RT-PCR experiment was repeated using pCAGGS vector-specific forward primer and hVEGF gene-specific reverse primer. In this case, both non-transfected islets and the islets transfected with LipofectAMINE/pCMS-EGFP complexes showed negligible amplification of hVEGF. On glucose challenge, insulin release from LipofectAMINE/pCAGGS-hVEGF transfected human islets increased from 10.78 ± 4.56 to 65 ± 5 ng/ml, suggesting little adverse effect on islet β cell response to glucose challenge. The low transfection efficiency is due to the islets being a cluster of approximately 1000 non-dividing cells. This underscores the importance of experimentation with the actual human islets.

Original languageEnglish (US)
Pages (from-to)89-100
Number of pages12
JournalMolecular Therapy
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2003

Keywords

  • Cationic liposomes
  • Green fluorescent protein
  • HVEGF
  • Human islets
  • Insulin secrection
  • Polymer
  • Real time RT-PCR
  • Transfection

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

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  • Cite this

    Mahato, R. I., Henry, J., Narang, A. S., Sabek, O., Fraga, D., Kotb, M., & Gaber, O. (2003). Cationic lipid and polymer-based gene delivery to human pacreatic islets. Molecular Therapy, 7(1), 89-100. https://doi.org/10.1016/S1525-0016(02)00031-X