Delivery of antisense oligonucleotides to leukemia cells by RNA bacteriophage capsids.
BACKGROUND
Acute myelogenous leukemia (AML) is the most common type of acute leukemia in adults. Because conventional treatments are associated with substantial side effects, novel therapeutic strategies are required. Antisense oligodeoxynucleotides (ODNs) have shown promise as the basis of emerging therapies for fighting cancer, although in vivo application is hampered by high sensitivity to cellular nuclease degradation. Encapsidation of ODNs in a drug-delivery capsule would reduce such degradation, thereby increasing the potency of therapy. MS2 bacteriophage capsid proteins may be used as novel virus-like particles (VLPs) to deliver ODNs. Here we report an analysis of the uptake mechanism of this VLP system and preliminary examples of its use to deliver a number of ODNs, including some targeting p120 messenger RNAs, a biomarker overexpressed in myelogenous leukemia cells.


METHODS
The ODNs were synthesized as covalent extensions to the translational repressor/assembly initiation signal (TR), a 19 nt stem-loop, of the RNA phage MS2. The VLPs were constructed by soaking ODN-TR directly into recombinant RNA-free capsid shells. Targeting of the encapsidated ODNs into cells was achieved by a receptor-mediated endocytosis pathway identified by immunofluorescence microscopy or by transmission electron microscopy with gold-labeled antibodies.


RESULTS
After covalent decoration with transferrin on their surface, the VLPs containing ODNs demonstrated increased effectiveness in killing target leukemia cells expressing transferrin receptors, suggesting that this system is worthy of more extensive analysis.


CONCLUSIONS
The findings suggest that RNA phage VLPs may be useful as a new nanomaterial for targeted delivery of antisense ODNs, or other macromolecular drug candidates.
Wu
Min
M
Department of Biochemistry and Molecular Biology, University of North Dakota, Grand Forks, North Dakota, USA.
Sherwin
Trevor
T
Brown
William L
WL
Stockley
Peter G
PG
D002213:Capsid / Q000737:chemistry*; D045744:Cell Line, Tumor; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry*; D018390:Gene Targeting / Q000379:methods; D018014:Gene Transfer Techniques; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D020320:Oligoribonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics*; D012317:RNA Phages / Q000378:metabolism*
D004337:Drug Carriers; D020320:Oligoribonucleotides, Antisense