Potent siRNA inhibitors of ribonucleotide reductase subunit RRM2 reduce cell proliferation In vitro and In vivo

Jeremy D. Heidel, Joanna Yi Ching Liu, Yun Yen, Bingsen Zhou, Bret S E Heale, John J. Rossi, Derek W. Bartlett, Mark E. Davis

Research output: Contribution to journalArticle

121 Citations (Scopus)

Abstract

Purpose: Ribonucleotide reductase (RR) is a therapeutic target for DNA replication - dependent diseases such as cancer. Here, a potent small interfering RNA (siRNA) duplex against the M2 subunit of RR (RRM2) is developed and shown to reduce the growth potential of cancer cells both in vitro and in vivo. Experimental Design: Three anti-RRM2 siRNAs were identified via computational methods, and the potency of these and additional "tiling" duplexes was analyzed in cultured cells via cotransfections using a RRM2-luciferase fusion construct. Knockdown of RRM2 by the best duplex candidates was confirmed directly by Western blotting. The effect of potent duplexes on cell growth was investigated by a real-time cell electronic sensing assay. Finally, duplex performance was tested in vivo in luciferase-expressing cells via whole animal bioluminescence imaging. Results: Moderate anti-RRM2 effects are observed from the three duplexes identified by computational methods. However, the tiling experiments yielded an extremely potent duplex (siR2B+5). This duplex achieves significant knockdown of RRM2 protein in cultured cells and has pronounced antiproliferative activity. S.c. tumors of cells that had been transfected with siR2B+5 preinjection grew slower than those of control cells. Conclusions: An anti-RRM2 siRNA duplex is identified that exhibits significant antiproliferative activity in cancer cells of varying human type and species (mouse, rat, monkey); these findings suggest that this duplex is a promising candidate for therapeutic development.

Original languageEnglish
Pages (from-to)2207-2215
Number of pages9
JournalClinical Cancer Research
Volume13
Issue number7
DOIs
Publication statusPublished - Apr 1 2007
Externally publishedYes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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    Heidel, J. D., Liu, J. Y. C., Yen, Y., Zhou, B., Heale, B. S. E., Rossi, J. J., Bartlett, D. W., & Davis, M. E. (2007). Potent siRNA inhibitors of ribonucleotide reductase subunit RRM2 reduce cell proliferation In vitro and In vivo. Clinical Cancer Research, 13(7), 2207-2215. https://doi.org/10.1158/1078-0432.CCR-06-2218