Potential gene therapy using e2f decoy approach with hvj-liposome method to glomerulonephritis in rats

N. Tomita*, G. H. Gibbons, J. Kim, S. Tomita, L. Zhang, Y. Kaneda, D. Barant, V. J. Dzau

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Mesangial cell (MC) proliferation is a central feature of the various renal diseases. The process of MC proliferation is dependent on the coordinated activation of a series of cell cycle regulatory genes. The transcriptional factor E2F plays an important role in the coordinated transactivation of cell cycle regulatory genes. We attempted to inhibit MC proliferation in a rat model of glomerulonephritis induced by anti-Thy 1.1 antibody by introduction of synthetic double stranded DNA with high affinity for E2F using HVJliposome method. First we confirmed that with HVJ-liposome method FITC-labeled oligonucleotide was able to be efficiently introduced into rat glomerular cells via renal artery. The anti-Thy 1.1 model was induced by systemic injection of monoclonal anti-Thy 1.1 antibody. After 30 minutes, HVJ-liposome solution with E2F decoy was injected into the left kidney via renal artery. On day 2, kidneys were removed to extract mRNA and reverse transcriptase polymerase chain reaction (RT-PCR) was performed. On day 5, cell proliferation was determined by 5′-bromo-2′-deoxy-uridine (BrdU) labeling method. Also cellularity was counted by using 3D analysis at the same time. The mRNA expressions of proliferating cell nuclear antigen (PCNA) and cdk2 kinase were decreased by the introduction of E2F decoy not by missense nor scrambled decoy. In the analysis of BrdU staining, there was a significant difference between left and right kidney. We also detected therapeitic effect in E2F treated rats compared to those treated with missense or scrambled decoy. Cellularity countings were associated with these RT-PCR or BrdU staining results. These evidences suggest that intrarenal delivery of E2F cis element decoy by HVJ-liposome method prevents the induction of cell cycle gene expression and MC proliferation. This study may have broad implication for the development of novel therapeutic approaches to glomerular diseases.

Original languageEnglish (US)
Pages (from-to)319a
JournalJournal of Investigative Medicine
Volume44
Issue number3
StatePublished - 1996

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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