Identification of benzodiazepine Ro5-3335 as an inhibitor of CBF leukemia through quantitative high throughput screen against RUNX1-CBFbeta interaction
- Authors
- 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.
- ID
- ZDB-PUB-120823-8
- Date
- 2012
- Source
- 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)
- Keywords
- none
- MeSH Terms
-
- Hematopoiesis/drug effects
- Zebrafish
- Molecular Sequence Data
- Blotting, Western
- Electrophoretic Mobility Shift Assay
- Histological Techniques
- Core Binding Factor Alpha 2 Subunit/metabolism*
- Mice
- Fluorescence Resonance Energy Transfer/methods
- Core Binding Factor beta Subunit/genetics
- Core Binding Factor beta Subunit/metabolism*
- Jurkat Cells
- Amino Acid Sequence
- Animals
- Humans
- Genetic Vectors/genetics
- Leukemia, Myeloid, Acute/genetics*
- Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics*
- Transcriptional Activation/drug effects*
- High-Throughput Screening Assays/methods*
- Protein Interaction Mapping/methods
- Flow Cytometry
- Surface Plasmon Resonance
- Benzodiazepines/pharmacology*
- Immunoprecipitation
- PubMed
- 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.