Use of a chemically modified antisense oligonucleotide library to identify and validate Eg5 (kinesin-like 1) as a target for antineoplastic drug development.
A library of 2'-methoxyethyl-modified antisense oligonucleotides (2'MOE ASO) targeting 1,510 different genes has been developed, validated, and used to identify cell cycle regulatory genes. The most effective molecular target identified was Eg5 (kinesin-like-1), which when inhibited gave the largest increase in 4N DNA in various tumor cells. The Eg5 ASO reduced Eg5 levels, inhibited proliferation, increased apoptosis, and altered the expression of other cell cycle proteins, including survivin and Aurora-A. To examine the therapeutic utility of the Eg5 ASO, the compound was also evaluated in xenograft models. Treatment with Eg5 ASO produced a statistically significant reduction of tumor growth, reduction in Eg5 expression in the tumors, and changes in histone phosphorylation, consistent with a loss of Eg5 protein expression. These data show, for the first time, the utility of a 2'MOE ASO library for high-throughput cell culture-based functional assays and suggest that an Eg5 ASO also has potential in a therapeutic strategy.
Department of Functional Genomics, Isis Pharmaceuticals, Carlsbad, California 92008, USA. ekoller@isisph.com
Erich
E
Koller
Stephanie
S
Propp
Hong
H
Zhang
Chenguang
C
Zhao
Xiaokun
X
Xiao
MingYi
M
Chang
Scott A
SA
Hirsch
Peter J
PJ
Shepard
Seongjoon
S
Koo
Cain
C
Murphy
Robert I
RI
Glazer
Nicholas M
NM
Dean
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001943:Breast Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D048708:Cell Growth Processes / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000235:genetics; D015723:Gene Library; D005910:Glioma / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000494:pharmacology*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; C470243:KIF11 protein, human; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D016547:Kinesins