Intravenous siRNA of Brain Cancer with Receptor Targeting and Avidin–Biotin Technology
- 681 Downloads
The effective delivery of short interfering RNA (siRNA) to brain following intravenous administration requires the development of a delivery system for transport of the siRNA across the brain capillary endothelial wall, which forms the blood–brain barrier in vivo.
siRNA was delivered to brain in vivo with the combined use of a receptor-specific monoclonal antibody delivery system, and avidin–biotin technology. The siRNA was mono-biotinylated on either terminus of the sense strand, in parallel with the production of a conjugate of the targeting MAb and streptavidin.
Rat glial cells (C6 or RG-2) were permanently transfected with the luciferase gene, and implanted in the brain of adult rats. Following the formation of intra-cranial tumors, the rats were treated with a single intravenous injection of 270 μg/kg of biotinylated siRNA attached to a transferrin receptor antibody via a biotin–streptavidin linker. The intravenous administration of the siRNA caused a 69–81% decrease in luciferase gene expression in the intracranial brain cancer in vivo.
Brain delivery of siRNA following intravenous administration is possible with siRNAs that are targeted to brain with the combined use of receptor specific antibody delivery systems and avidin–biotin technology.
Key wordsavidin biotin blood–brain barrier monoclonal antibody RNA interference
- 9.W. M. Pardridge, Y. S. Kang, J. L. Buciak, J. Yang. Human insulin receptor monoclonal antibody undergoes high affinity binding to human brain capillaries in vitro and rapid transcytosis through the blood–brain barrier in vivo in the primate. Pharm. Res. 12:807–816 (1995).PubMedCrossRefGoogle Scholar
- 20.I. Pilz, E. Schwarz, W. Durchschein, A. Licht, M. Sela. Effect of cleaving interchain disulfide bridges on the radius of gyration and maximum length of anti-poly(d-alanyl) antibodies before and after reaction with tetraalanine hapten. Proc. Natl. Acad. Sci. USA. 77:117–121 (1980).PubMedCrossRefGoogle Scholar
- 21.M. Grzelinski, B. Urban-Klein, T. Martens, K. Lamszus, U. Bakowsky, S. Hobel, F. Czubayko, A. Aigner. RNA interference-mediated gene silencing of pleiotrophin though polyethylenimine-complexed small interfering RNAs in vivo exerts antitumoral effects in glioblastoma xenografts. Hum. Gene Ther. 17:751–766 (2006).PubMedCrossRefGoogle Scholar
- 28.M. Egli, G. Minasov, V. Tereshko, P. S. Pallan, M. Teplova, G. B. Inamati, E. A. Lesnik, S. R. Owens, B. S. Ross, T. P. Prakash, M. Manoharan. Probing the influence of stereoelectronic effects on the biophysical properties of oligonucleotides: comprehensive analysis of the RNA affinity, nuclease resistance, and crystal structure of ten 2-O-ribonucleic acid modifications. Biochemistry 44:9045–9057 (2005).PubMedCrossRefGoogle Scholar
- 29.T. Yoshikawa, W. M. Pardridge. Biotin delivery to brain with a covalent conjugate of avidin and a monoclonal antibody to the transferrin receptorJ. Pharmacol. Exp. Ther. 263:897–903 (1994).Google Scholar
- 30.M. L. Penichet, Y. S. Kang, W. M. Pardridge, S. L. Morrison, S.U. Shin. An antibody-avidin fusion protein specific for the transferrin receptor serves as a delivery vehicle for effective brain targeting: initial applications in anti-HIV antisense drug delivery to the brain. J. Immunol. 163:4421–4426 (1999).PubMedGoogle Scholar