Complete in vivo reversal of the multidrug resistance phenotype by jet-injection of anti-MDR1 short hairpin RNA-encoding plasmid DNA.

Abstract

Triggering the RNA interference (RNAi) pathway by inducing the expression of short hairpin RNA (shRNA) molecules has become a promising tool for efficient silencing of a given gene in gene therapy applications. In this study, shRNA encoding DNA was utilized to reverse the classical MDR1/P-glycoprotein (MDR1/P-gp)-mediated multidrug resistance (MDR) phenotype in vivo. For the first time, the nonviral jet-injection technology was applied for delivering naked shRNA-vector constructs for direct intratumoral in vivo transfer. The highly efficient anti-MDR1 shRNA expression vectors were applied twice in the human MDR1/P-gp overexpressing MaTu/ADR cancer xenograft-bearing mice, and twice in the corresponding drug-sensitive parental MaTu tumor xenograft bearing mice as well. Two days after anti-MDR1 shRNA vector injection, the expression level of the MDR1 messenger RNA (mRNA) was decreased by more than 90% and the corresponding MDR1/P-gp protein was no longer detectable in the tumors. Two jet-injections of anti-MDR1 shRNA vectors into the tumors, combined with two intravenous (IV) administrations of doxorubicin, were sufficient to achieve complete reversal of the drug-resistant phenotype. The data show that jet-injection delivery of shRNA-expressing vectors is effective in reversing MDR1/P-gp-mediated MDR in vivo, and is therefore a promising strategy for making tumors with an MDR1/Pgp-dependent MDR phenotype revert to a drug-sensitive state.

• Authors: Ulrike Stein, Wolfgang Walther, Alexandra Stege, Alexander Kaszubiak, Iduna Fichtner, Hermann Lage
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 16 Issue 1 Pg. 178-86 (Jan 2008) ISSN: 1525-0024 [Electronic] United States
• PMID: 17878902 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• ABCB1 protein, human
• ATP Binding Cassette Transporter, Subfamily B
• ATP Binding Cassette Transporter, Subfamily B, Member 1
• DNA, Neoplasm
• RNA, Messenger
• RNA, Neoplasm

Topics

• ATP Binding Cassette Transporter, Subfamily B
• ATP Binding Cassette Transporter, Subfamily B, Member 1 (antagonists & inhibitors, biosynthesis, genetics)
• Animals
• DNA, Neoplasm (administration & dosage, genetics)
• Drug Resistance, Multiple (genetics)
• Drug Resistance, Neoplasm (genetics)
• HeLa Cells
• Humans
• Injections, Intralesional
• Injections, Jet
• Mammary Neoplasms, Experimental (genetics, metabolism, therapy)
• Mice
• Mice, Nude
• Phenotype
• Plasmids (administration & dosage)
• RNA, Messenger (antagonists & inhibitors, biosynthesis)
• RNA, Neoplasm (administration & dosage, genetics)