Identification of benzodiazepine Ro5-3335 as an inhibitor of CBF leukemia through quantitative high throughput screen against RUNX1-CBFbeta interaction

Cunningham, L., Finckbeiner, S., Hyde, R.K., Southall, N., Marugan, J., Yedavalli, V.R., Dehdashti, S.J., Reinhold, W.C., Alemu, L., Zhao, L., Yeh, J.R., Sood, R., Pommier, Y., Austin, C.P., Jeang, K.T., Zheng, W., and Liu, P.
Proceedings of the National Academy of Sciences of the United States of America   109(36): 14592-14597 (Journal)
Registered Authors
Liu, Pu Paul, Sood, Raman, Yeh, Jing-Ruey (Joanna)
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Benzodiazepines/pharmacology*
  • Blotting, Western
  • Core Binding Factor Alpha 2 Subunit/metabolism*
  • Core Binding Factor beta Subunit/genetics
  • Core Binding Factor beta Subunit/metabolism*
  • Electrophoretic Mobility Shift Assay
  • Flow Cytometry
  • Fluorescence Resonance Energy Transfer/methods
  • Genetic Vectors/genetics
  • Hematopoiesis/drug effects
  • High-Throughput Screening Assays/methods*
  • Histological Techniques
  • Humans
  • Immunoprecipitation
  • Jurkat Cells
  • Leukemia, Myeloid, Acute/genetics*
  • Mice
  • Molecular Sequence Data
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics*
  • Protein Interaction Mapping/methods
  • Surface Plasmon Resonance
  • Transcriptional Activation/drug effects*
  • Zebrafish
22912405 Full text @ Proc. Natl. Acad. Sci. USA

Core binding factor (CBF) leukemias, those with translocations or inversions that affect transcription factor genes RUNX1 or CBFB, account for <24% of adult acute myeloid leukemia (AML) and 25% of pediatric acute lymphocytic leukemia (ALL). Current treatments for CBF leukemias are associated with significant morbidity and mortality, with a 5-y survival rate of <50%. We hypothesize that the interaction between RUNX1 and CBFβ is critical for CBF leukemia and can be targeted for drug development. We developed high-throughput AlphaScreen and time-resolved fluorescence resonance energy transfer (TR-FRET) methods to quantify the RUNX1–CBFβ interaction and screen a library collection of 243,398 compounds. Ro5-3335, a benzodiazepine identified from the screen, was able to interact with RUNX1 and CBFβ directly, repress RUNX1/CBFB-dependent transactivation in reporter assays, and repress runx1-dependent hematopoiesis in zebrafish embryos. Ro5-3335 preferentially killed human CBF leukemia cell lines, rescued preleukemic phenotype in a RUNX1–ETO transgenic zebrafish, and reduced leukemia burden in a mouse CBFB–MYH11 leukemia model. Our data thus confirmed that RUNX1–CBFβ interaction can be targeted for leukemia treatment and we have identified a promising lead compound for this purpose.

Genes / Markers
Show all Figures
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes