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BioAssay: AID 651556

qHTS Assays for the Identification of Compounds that Synergize with a BTK Inhibitor (Ibrutinib, PCI-32765)

In recent years, using advanced gene expression profiling techniques the laboratory of Dr. Louis Staudt at NCI/NIH has revealed that a class of lymphoma referred to as Diffuse Large B-Cell Lymphoma (DLBCL) is composed of at least 3 different sub-groups, each having distinct oncogenic mechanisms that respond differently to therapies. The sub-group classified as activated B cell-like diffuse large more ..
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 Related BioAssays
 Related BioAssays
AID: 651556
Data Source: NCGC (MIPE3-NFKB-Summary)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2012-08-30
Modify Date: 2014-01-07
Depositor Specified Assays
AIDNameTypeComment
651645Cell Proliferation Assay against the TMD8 Cell Lineconfirmatory
651646Cell Proliferation Assay against the HBL1 Cell Lineconfirmatory
624476Cytotoxicity counterscreen for NFkB agonists and antagonistsconfirmatory
624479qHTS Assay for Identification of Small Molecule Antagonists for NFkB Signaling Pathway.confirmatory
624478qHTS Assay for Identification of Small Molecule Agonists for NFkB Signaling Pathway.confirmatory
651713Cell Proliferation Assay against the TMD8 Cell Line (Caspase readout at 8 hrs)confirmatory
651712Cell Proliferation Assay against the TMD8 Cell Line (Caspase readout at 16 hrs)confirmatory
Description:
In recent years, using advanced gene expression profiling techniques the laboratory of Dr. Louis Staudt at NCI/NIH has revealed that a class of lymphoma referred to as Diffuse Large B-Cell Lymphoma (DLBCL) is composed of at least 3 different sub-groups, each having distinct oncogenic mechanisms that respond differently to therapies. The sub-group classified as activated B cell-like diffuse large B cell lymphoma (ABC DLBCL) demonstrates activation of B-cell receptors and the NFkappaB signaling pathway. One class of targeted therapeutics showing early clinical success in killing of ABC DLBCL lines are small molecule inhibitors of the Bruton tyrosine kinase (BTK) 11. Thus, we chose to investigate what additional compounds, if any, would synergize with the BTK inhibitor Ibrutinib (PCI-32765).

Prior to any combination assays, using an NCATS developed oncology compound library called the Mechanism of Interrogation Plate (MIPE), two well studied ABC DLBCL lines (TMD8 and HBL1) were screened using a cell viability assay (CellTiterGlo) and a 12-point titration series for each single agent. These initial experiments offered insight into the concentration range that offered the best coverage of activity and non-activity to assure relevant data is generated. In addition to the viability signature associated with this library, we examined the apoptotic response of each agent versus the TMD8 line as judged by caspase 3/7 activation. Activity was assayed at both 8 and 16 hour time-points to gain an appreciation of which agents induced an apoptotic response in a reasonable timeframe from a therapeutic exposure perspective. MIPE was also assayed for single agent viability in a human mesenchymal stem cell (hMSC) line as a surrogate of each library member's potential therapeutic index.

The combination plates were then generated by choosing 20 of the best compounds from this single agent screen and plated in the presence of the BTK inhibitor. The first analysis was carried out using 6x6 dose response blocks for each of the 20 compounds. Compounds that demonstrated synergy in this 6x6 assay were then selected for follow-ups in a more extensive 10x10 dose response format. In addition to the cell viability assay, the parallel caspase 3/7 activation and viability in hMSC assays were carried out in the 10x10 format.
Protocol
Please see linked AIDs for a detailed protocol.
Comment
This is the summary AID for "qHTS Assays for the Identification of Compounds that Synergize with a BTK Inhibitor (Ibrutinib, PCI-32765)"
Additional Information
Grant Number: MH084681 U54

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