Therapeutic potentialities of EWS-Fli-1 mRNA-targeted vectorized antisense oligonucleotides.
We have used structured antisense oligonucleotides (AON), which are protected against extra and intracellular degradation by their internal structure. We have shown that if correctly designed this structure does not prevent them from hybridizing to the mRNA target. This concept allows reducing the number of thioate groups in the oligonucleotide and therefore the potential toxicity. Junction oncogenes are found in cancers such as certain leukemias, Ewing sarcoma, and thyroid papillary carcinomas. Ewing sarcoma is a cancer of children and young adults with bone metastasis. It is caused by a chromosomic translocation t(11;22) (q24;q12) creating a fusion gene between the genes EWS and Fli-1 giving rise to a chimeric protein which is an unnatural transcription factor. Immortalized NIH/3T3 cells transfected by the EWS-Fli-1 cDNA under the control of the LTR retroviral promoter--which do not undergo apoptosis and which became tumoral--were used for this study. As a model of Ewing sarcoma in nude mice, we have used permanently expressing human EWS-Fli-1 cells grafted to nude mice. The nanospheres or nanocapsules have been used to deliver two different AON: a phosphorothioate, and a structured chimeric AON, both targeted toward the junction area of EWS-Fli-1. Both types of AON-loaded nanoparticles inhibited the growth of the xenografted tumor after intratumoral injections into nude mice, whereas similar nanoparticles with control oligonucleotides had no effect. With AON in nanospheres, we have shown after 24 hours that the mRNA of EWS-Fli-1 was specifically down-regulated, confirming the antisense activity of the targeted AON.
Maksimenko
A
A
BioAlliance Pharma, 59, Boulevard du Général Martial Valin, 75015 Paris.
Lambert
G
G
Bertrand
J R
JR
Fattal
E
E
Couvreur
P
P
Malvy
C
C
D000818:Animals; D005822:Genetic Vectors; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000188:drug therapy*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D015514:Oncogene Proteins, Fusion / Q000096:biosynthesis / Q000235:genetics*; D051789:Proto-Oncogene Protein c-fli-1; D034622:RNA Interference / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects*; D034802:RNA-Binding Protein EWS; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*
C106031:EWS-FLI fusion protein; D016376:Oligonucleotides, Antisense; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D012333:RNA, Messenger; D034802:RNA-Binding Protein EWS; D014157:Transcription Factors