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BioAssay: AID 504373

qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: Zebrafish assay for Hematopoiesis, SAR for Probe

Core Binding Factor (CBF) abnormalities are associated with 20-25% of all acute myeloid leukemias (AML), of which 5-10% are further sub classified as acute myelomonocytic leukemia with eosinophilia, also known as M4Eo in the FAB classification scheme. This subtype of leukemia is usually associated with chromosome 16 inversion (p13:q22). Chromosome 16 inversion results in formation of the fusion more ..
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 Tested Compounds
 Tested Compounds
All(26)
 
 
Active(6)
 
 
Inactive(20)
 
 
Inconclusive(1)
 
 
 Tested Substances
 Tested Substances
All(27)
 
 
Active(6)
 
 
Inactive(20)
 
 
Inconclusive(1)
 
 
AID: 504373
Data Source: NCGC (CBFB010)
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-02-25
Hold-until Date: 2011-08-25
Modify Date: 2011-10-03

Data Table ( Complete ):           Active    All
Targets
BioActive Compounds: 6
Depositor Specified Assays
AIDNameTypeComment
1477qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid LeukemiaconfirmatoryPrimary screen.
1484Quantitative High-Throughput Screen for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: SummarysummarySummary assay for project.
Description:
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Production centers Network [MLPCN]

MLPCN Grant: X01MH083259-01
Assay Submitter (PI): Paul Liu

NCGC Assay Overview:

Core Binding Factor (CBF) abnormalities are associated with 20-25% of all acute myeloid leukemias (AML), of which 5-10% are further sub classified as acute myelomonocytic leukemia with eosinophilia, also known as M4Eo in the FAB classification scheme. This subtype of leukemia is usually associated with chromosome 16 inversion (p13:q22). Chromosome 16 inversion results in formation of the fusion oncogene CBFB-MYH11, which encodes the fusion protein CBF-beta-SMMHC. This fusion protein binds to RUNX1 (AML1) with high affinity and dominantly inhibits RUNX1 function preventing definitive hematopoiesis.

It has been hypothesized that the interference of CBF-beta and RUNX1 binding could have therapeutic implications for patients with CBF mediated leukemias. We developed a CBF-beta and RUNX1 binding assay in AlphaScreenTM format and optimized it for high throughput screening to search for inhibitors. The goal of this project is to screen MLPCN's compound collection (MLSMR) for identifying the inhibitors of the interaction between CBF-beta and RUNX1 proteins as research probes. If these probes work well in the animal models, the ultimate goal of this project is to development a new drug treatment for this disease.

The present assay assesses the ability of compounds to selectively inhibit hematopoiesis in a zebrafish whole organism assay. These fish contain a genomic modulation where GFP has been fused to cd41, a marker for thrombocytes, and thus circulating thrombocytes can be easily imaged in anesthetized fish.
Protocol
Assay protocol:

GFP-cd41 zebrafish were kindly provided by the Liu lab. Zebrafish embryos were incubated with compound (<0.1% DMSO final) at 48 hours post fertilization, for 5 days. Embryos were manually scored under a microscope with GFP illumination for circulating GFP-tagged thrombocytes.

Keywords: MLSMR, MLPCN, NIH Roadmap, qHTS, NCGC, Core binding factor, CBF, CBF-beta, Acute myeloid leukemia
Comment
Compounds were scored based on their ability to reduce the number of circulating GFP-tagged thrombocytes without causing obvious harm to the embryo. Compounds that were inactive were given a score of 0. Compounds harmful to the embryo were given a score of 10. Active compounds were given a score of 50.
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Embryo conditionTextual description of embryo condition after incubation with compound for three days.String
2Activity at 1 uM (1μM**)Percent of fish with circulating GFP cells at given concentration of compound.Float%
3Activity at 10 uM (10μM**)Percent of fish with circulating GFP cells at given concentration of compound.Float%
4Activity at 25 uM (25μM**)Percent of fish with circulating GFP cells at given concentration of compound.Float%
5Activity at 50 uM (50μM**)Percent of fish with circulating GFP cells at given concentration of compound.Float

** Test Concentration.
Additional Information
Grant Number: X01MH083259-01

Data Table (Concise)
Classification
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