Identification of benzodiazepine Ro5-3335 as an inhibitor of CBF leukemia through quantitative high throughput screen against RUNX1-CBFβ interaction.
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.
Oncogenesis and Development Section, National Institute of Allergy and Infectious Diseases, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Lea
L
Cunningham
Steven
S
Finckbeiner
R Katherine
RK
Hyde
Noel
N
Southall
Juan
J
Marugan
Venkat R K
VR
Yedavalli
Seameen Jean
SJ
Dehdashti
William C
WC
Reinhold
Lemlem
L
Alemu
Ling
L
Zhao
Jing-Ruey Joanna
JR
Yeh
Raman
R
Sood
Yves
Y
Pommier
Christopher P
CP
Austin
Kuan-Teh
KT
Jeang
Wei
W
Zheng
Paul
P
Liu
D000595:Amino Acid Sequence; D000818:Animals; D001569:Benzodiazepines / Q000494:pharmacology*; D015153:Blotting, Western; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism*; D050658:Core Binding Factor beta Subunit / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D005434:Flow Cytometry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006410:Hematopoiesis / Q000187:drug effects; D057166:High-Throughput Screening Assays / Q000379:methods*; D006652:Histological Techniques; D006801:Humans; D047468:Immunoprecipitation; D019169:Jurkat Cells; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D025941:Protein Interaction Mapping / Q000379:methods; D020349:Surface Plasmon Resonance; D015533:Transcriptional Activation / Q000187:drug effects*; D015027:Zebrafish
C493693:Cbfb protein, mouse; D050676:Core Binding Factor Alpha 2 Subunit; D050658:Core Binding Factor beta Subunit; C493726:Runx1 protein, mouse; D001569:Benzodiazepines