Systemic delivery of HK Raf-1 siRNA polyplexes inhibits MDA-MB-435 xenografts.
Our past research has focused on identifying an effective carrier composed of histidine and lysine for delivery of nucleic acid into cells. For this purpose, we developed histidine-lysine-rich (HK) polymers with specific sequences and branching. We have found that branched HK polymers in complex with Raf-1 siRNA markedly decreased Raf-1 mRNA and induced apoptosis in cell lines in vitro. The primary focus of the present study was to determine an effective carrier to deliver siRNA systemically to tumor xenografts. After comparing HK:Raf-1 polyplexes for their in-vivo efficacy, we investigated in greater detail whether one of these polymers, H3K(+H)4b, in complex with Raf-1 siRNA, inhibited the growth of MDA-MB-435 xenografts. H3K(+H)4b is a four-branched HK peptide whose predominant repeating sequence within the terminal arm is -HHHK-. After the first tail-vein injection in a mouse model, there was a statistically significant reduction in tumor size between the H3K(+H)4b:Raf-1 siRNA-treated and the control groups (P<0.01). By the third injection, there was nearly a 50% reduction in the Raf-1 siRNA-treated group compared to the control siRNA-treated or -untreated group. Consistent with a significant effect of the HK:Raf-1 polyplex on the tumor, there were marked histological changes, increased apoptosis and fewer vessels in the Raf-1 siRNA-treated group. Raf-1 protein within the tumor was significantly decreased after treatment with the HK:Raf-1 siRNA polyplex compared to the control treatment groups. Despite the striking effect on the tumor by the HK Raf-1 siRNA, there was little evidence of toxicity in normal tissues with this therapy. By harnessing the ability to modify the amino-acid sequence and branching of HK polymers, we expect continued development of HK polymers as in-vivo carriers of siRNA.
Leng
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Department of Pathology, University of Maryland, Baltimore, MD 21201, USA.
Scaria
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Lu
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Woodle
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MC
Mixson
A J
AJ
D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000378:metabolism; D008239:Lysine / Q000737:chemistry*; D051379:Mice; D008819:Mice, Nude; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection; D014183:Transplantation, Heterologous
D019394:Ki-67 Antigen; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D034741:RNA, Small Interfering; D006639:Histidine; D019908:Proto-Oncogene Proteins c-raf; D008239:Lysine