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BioAssay: AID 651869

Cidal vs. static assay: inhibition of T. cruzi (CAI/72) after prolonged treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-08_Inhibitor_SinglePoint_DryPowder_Activity_Set2

Assay Overview: This assay was conducted to determine if the compounds are cidal to Trypanosoma cruzi cells (amastigotes) within the mammalian host cell. Bovine Embryo Skeletal Muscle Cells (BESM)infected with CAI/72 strain of Trypanosoma cruzi. After 24 hours, cells were washed and compounds were added fresh ever other day for 29 days. After compound treatment, compound was removed from the more ..
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 Tested Compounds
 Tested Compounds
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Active(10)
 
 
 Tested Substances
 Tested Substances
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Active(10)
 
 
 Related BioAssays
 Related BioAssays
AID: 651869
Data Source: Broad Institute (2138-08_Inhibitor_SinglePoint_DryPowder_Activity_Set2)
Depositor Category: NIH Molecular Libraries Probe Production Network, Assay Provider
BioAssay Version:
Deposit Date: 2012-12-06
Hold-until Date: 2013-07-15
Modify Date: 2013-07-16

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 10
Related Experiments
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AIDNameTypeProbeComment
624280Broad Institute Inhibition of T.cruzi replication in culture Inhibitor Probe ProjectSummary1 depositor-specified cross reference: Summary Assay
624255Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_SinglePoint_HTS_ActivityScreening same project related to Summary assay
651739Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_SinglePoint_CherryPick_ActivityOther same project related to Summary assay
651740Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_SinglePoint_CherryPick_Activity_Set2Other same project related to Summary assay
651742NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_SinglePoint_CherryPick_ActivityOther same project related to Summary assay
651744NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_SinglePoint_CherryPick_Activity_Set2Other same project related to Summary assay
651817NIH/3T3 (mouse embryonic fibroblast ) toxicity Measured in Cell-Based System Using Plate Reader - 2017-02_Inhibitor_Dose_CherryPick_Activity_Set2Confirmatory same project related to Summary assay
651818Intracellular Trypanosomes Measured in Cell-Based/Microorganism System Using Plate Reader - 2017-01_Inhibitor_Dose_CherryPick_Activity_Set3Confirmatory same project related to Summary assay
651844NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651845Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651877Cidal vs. static assay: inhibition of T. cruzi (CAI/72) after prolonged treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-08_Inhibitor_SinglePoint_DryPowder_ActivityOther same project related to Summary assay
651881Inhibition of T. cruzi Tulahuen strain amastigotes in L-6 muscle cells Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2138-03_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651885Inhibition of intracellular CAI/72 strain of Trypanosoma cruzi in bovine embryo skeletal muscle cells: high content imaging Measured in Cell-Based System Using Imaging - 2138-07_Inhibitor_SinglePoint_DryPowder_ActivityConfirmatory same project related to Summary assay
651888Inhibition of Plasmodium falcipirum NF54 Measured in Cell-Based System Using Plate Reader - 2138-11_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651889Inhibition of Plasmodium falcipirum NF54 Measured in Cell-Based System Using Plate Reader - 2138-11_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651890Inhibition of T. cruzi Tulahuen strain amastigotes in L-6 muscle cells Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2138-03_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651891Inhibition of L. donovani axenic amastigotes Measured in Cell-Based System Using Plate Reader - 2138-04_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651892Inhibition of L. donovani axenic amastigotes Measured in Cell-Based System Using Plate Reader - 2138-04_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651893Inhibition of Trypanosoma brucei rhodesiense STIB900 Measured in Cell-Based System Using Plate Reader - 2138-05_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651894Inhibition of Trypanosoma brucei rhodesiense STIB900 Measured in Cell-Based System Using Plate Reader - 2138-05_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651895L-6 rat myocyte cell toxicity Measured in Cell-Based System Using Plate Reader - 2138-06_Inhibitor_Dose_DryPowder_ActivityConfirmatory same project related to Summary assay
651896L-6 rat myocyte cell toxicity Measured in Cell-Based System Using Plate Reader - 2138-06_Inhibitor_Dose_CherryPick_ActivityConfirmatory same project related to Summary assay
651897Inhibition of Trypanosoma cruzi Cruzain Measured in Biochemical System Using Plate Reader - 2138-09_Inhibitor_SinglePoint_DryPowder_ActivityConfirmatory same project related to Summary assay
651903Intracellular Trypanosomes Measured in Cell-Based/Microorganism System Using Plate Reader - 2017-01_Inhibitor_SinglePoint_HTS_Activity_Set2Screening same project related to Summary assay
652275Inhibition of T.cruzi proliferation in culture Measured in Cell-Based System Using Plate Reader - 2138-01_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
652278Cidal vs. static assay: inhibition of T. cruzi (CAI/72) in J774 macrophages after prolonged compound treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-13_Inhibitor_SinglePoint_DryPowder_Activity_Set2Other same project related to Summary assay
652279Cidal vs. static assay: inhibition of T. cruzi (CAI/72) in J774 macrophages after prolonged compound treatment and treatment withdrawal Measured in Cell-Based System Using Imaging - 2138-13_Inhibitor_SinglePoint_DryPowder_ActivityOther same project related to Summary assay
652280NIH/3T3 (mouse embryonic fibroblast) toxicity Measured in Cell-Based System Using Plate Reader - 2138-02_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatory same project related to Summary assay
686919Inhibition of Trypanosoma cruzi Cruzain Measured in Biochemical System Using Plate Reader - 2138-09_Inhibitor_Dose_DryPowder_ActivityOther same project related to Summary assay
Description:
Keywords: Trypanosoma cruzi, amastigotes, cidal


Assay Overview: This assay was conducted to determine if the compounds are cidal to Trypanosoma cruzi cells (amastigotes) within the mammalian host cell. Bovine Embryo Skeletal Muscle Cells (BESM)infected with CAI/72 strain of Trypanosoma cruzi. After 24 hours, cells were washed and compounds were added fresh ever other day for 29 days. After compound treatment, compound was removed from the infected cells and observed for 20 days. The emergence of fully developed trypomastigotes was assessed daily via light microscope. The mammalian T.cruzi life cycle is 6 days; when treated with DMSO or a nonactive compound, 6 days after infection is the earliest possible emergence of trypomastigotes. If a compound lyses all intracellular amastigotes, no trypomastigotes will emerge at any day during compound treatment or compound withdrawal. However, if a compound is static or only partially lytic, there will be a delay of trypomastigote emergence beyond the traditional 6 days. For graphic representation, day "0" is the first day of compound withdrawal. Compound treatment starts on day '-29'. Therefore the trypomastigotes appear at day '-23' when treated with DMSO only.


Expected Outcome: For those compounds that are cidal, no trypomastigotes will appear at any day tested with or without compound treatment. For those compounds that have a static effect, it is expected that compound treatment will suppress replication during treatment, but upon compound removal, trypomastigotes will immediately reappear. Compounds that alter the normal replication of T. cruzi may extend the normal life cycle past 6 days even after compound removal.
Protocol
This assay assesses cidal activity of compounds in T. cruzi (CAI/72 )-infected BESM cells (Bovine Embryo Skeletal Muscle Cells). Twenty four hours after infection, infected cells are washed and compounds are added every other day for 29 days. After the initial period, compounds are removed and cells are observed for an additional 20 days without drug, during which time cultures are monitored for reappearance of T. cruzi parasites. This procedure allows us to determine if host cells were effectively cured (i.e., compound is cidal) or if infection still persists (i.e, static activity).
In recent years the cidality assay has been used to evaluate over 1,000 compounds of various structural types. For the work proposed herein, we will test compounds in full dose response (10, 3.3, 1.1, 0.37, 0.12, and 0.04 microM test compound) so that meaningful comparison between analogs can be made (K777 is curative at 10 mM in this assay). Highly effective compounds can also be tested against a diverse panel of T. cruzi strains including CA-I/72, Y, SX10/7, MF (UCSF), BB (UCSF), and CL.
Comment
To analyze the data, the day of trypomastigote reemergence was added to a linear scale between -29 (1st day of compound addition) to 20 (final day of experiment). Concentrations vs. day of emergence were plotted in Graphpad prism and a linear regression was conducted to determine the slope. At the first concentration where no trypomastigotes appeared, the last day of the experiment (20) was added as a data point for the linear regression.
Pubchem Activity Score was calculated by dividing the slope of the test compound by the slope of Benznidazole, then multiplying by 10 to bring the values to a scale between 0 and 100. Larger numbers are more active and lower numbers are less active.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Format: Cell-based
Assay Cell Type: BESM
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Day_of_Emergence_at_10uM (10μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
2Day_of_Emergence_at_3.3uM (3.3μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
3Day_of_Emergence_at_1.1uM (1.1μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
4Day_of_Emergence_at_0.37uM (0.37μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
5Day_of_Emergence_at_0.12uM (0.12μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
6Day_of_Emergence_at_0.04uM (0.04μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
7Day_of_Emergence_at_0.0uM (0μM**)Number of days after final treatment at the indicated concentration at which parasite emergence was observedIntegerDays after withdrawal
8SlopeLinear regression of emergence dataFloat

** Test Concentration.
Additional Information
Grant Number: 1 R03 MH085673-01

Data Table (Concise)
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