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BioAssay: AID 488929

Elucidation of physiology of non-replicating, drug-tolerant Mycobacterium tuberculosis - Summary

Assay Rationale and Summary: Over one third of the world's population is infected with tuberculosis. In 1993, the World Health Organization declared a global health emergency with the resurgence of tuberculosis in the setting of HIV infection and with the emergence of multi-drug resistant TB. In 2004, 14.6 million people had active TB, 8.9 million new cases were documented, and 1.7 million more ..
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AID: 488929
Data Source: Southern Research Specialized Biocontainment Screening Center (Hung Summary)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-10-26
Modify Date: 2013-08-29

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compound: Chemical Probe: 1    Active: 1
Related Experiments
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AIDNameTypeComment
488890Elucidation of physiology of non-replicating, drug-tolerant Mycobacterium tuberculosisConfirmatorydepositor-specified cross reference: Primary screen and Confirmatory dose response of hits
489018A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Non-Replicating, Drug-tolerant Mycobacterium tuberculosisConfirmatorydepositor-specified cross reference: Hits from Primary screen in cytotoxicity dose response with HepG2 cells.
489025A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Non-Replicating, Drug-tolerant Mycobacterium tuberculosisConfirmatorydepositor-specified cross reference: Hits from Primary screen in cytotoxicity dose response with THP1 cells.
492952A Cell Based Secondary Assay to Explore Compounds that Modulate Non-Replicating, Drug-tolerant Compounds in Replicating H37Rv TB of Mycobacterium tuberculosisConfirmatorydepositor-specified cross reference
492998A Cell Based Secondary Assay to Explore Cytotoxicity in Vero E6 Cells of Compounds that Modulate Non-Replicating, Drug-tolerant Mycobacterium tuberculosisConfirmatorydepositor-specified cross reference: Hits from Primary screen in cytotoxicity dose response with Vero E6 cells.
651617Counterscreen for inhibitors of non replicating M. tb using log phase replicating mycobacteria Measured in Microorganism System Using Plate Reader - 2157-02_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference
651618Counterscreen for inhibitors of non replicating M. tb using log phase replicating mycobacteria Measured in Microorganism System Using Plate Reader - 2157-02_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorydepositor-specified cross reference
651619Secondary assay to identify inhibitors of non-replicating M. tb using luciferase expression without log phase outgrowth Measured in Microorganism System Using Plate Reader - 2157-04_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorydepositor-specified cross reference
651620Secondary assay to identify inhibitors of non-replicating M. tb using luciferase expression without log phase outgrowth Measured in Microorganism System Using Plate Reader - 2157-04_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference
651850Inhibition of Mycobacterium tuberculosis during logarithmic growth, as enumerated by colony forming units_2157-05_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference
651851Counterscreen for inhibitors of non replicating M. tb using log phase replicating mycobacteriaConfirmatorydepositor-specified cross reference
651856Secondary assay to identify inhibitors of non-replicating M. tb using luciferase expression without log phase outgrowthConfirmatorydepositor-specified cross reference
651857Inhibition of Mycobacterium tuberculosis during starvation growth, as enumerated by colony forming units_2157-06_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference
651946HeLa Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-04_Other_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference
651947HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Other_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference
651949HEK293 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-01_Other_Dose_DryPowder_Activity_Set6Confirmatorydepositor-specified cross reference
687033Broad Institute Elucidation of the physiology of non-replicating, drug tolerant Mycobacterium tuberculosis: Probe for both Non-Replicating & Replicating organisms Inhibitor Probe ProjectSummarydepositor-specified cross reference
687034Broad Institute Elucidation of the physiology of non-replicating, drug tolerant Mycobacterium tuberculosis: Probe to inhibit the Transition from Non-Replication to Replication Inhibitor Probe ProjectSummarydepositor-specified cross reference
743175Elucidation of physiology of non-replicating, drug-tolerant Mycobacterium tuberculosis Measured in Microorganism System Using Plate Reader - 2157-01_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
743183Secondary assay to identify inhibitors of non-replicating M. tb using luciferase expression without log phase outgrowth Measured in Microorganism System Using Plate Reader - 2157-04_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
743186TB:Log Assay, OD600 Measured in Microorganism System Using Plate Reader - 2157-02_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
743162TB: CFU Replicating Measured in Microorganism System Using Imaging - 7108-01_Inhibitor_Dose_DryPowder_ActivityOthersame project related to Summary assay
743163TB: CFU Starved Measured in Microorganism System Using Imaging - 7108-02_Inhibitor_Dose_DryPowder_ActivityOthersame project related to Summary assay
743164TB:CFU starved Measured in Microorganism System Using Imaging - 7108-04_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
743165TB: CFU replicating Measured in Microorganism System Using Imaging - 7108-03_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
743174TB: Log Assay, OD 600 Measured in Microorganism System Using Plate Reader - 7108-05_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
743175Elucidation of physiology of non-replicating, drug-tolerant Mycobacterium tuberculosis Measured in Microorganism System Using Plate Reader - 2157-01_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
743183Secondary assay to identify inhibitors of non-replicating M. tb using luciferase expression without log phase outgrowth Measured in Microorganism System Using Plate Reader - 2157-04_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
743186TB:Log Assay, OD600 Measured in Microorganism System Using Plate Reader - 2157-02_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
743404TB: CFU replicating Measured in Microorganism System Using Imaging - 7108-05_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
743450TB:CFU starved Measured in Microorganism System Using Imaging - 7108-06_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
743485On Hold
743486On Hold
1035471On Hold
1035472On Hold
1035473On Hold
Description:
Southern Research's Specialized Biocontainment Screening Center (SRSBSC)
Southern Research Institute (Birmingham, Alabama)
NIH Molecular Libraries Probe Production Centers Network (MLPCN)
Assay Provider: Dr. Deborah Hung, Massachusetts Institute of Technology
Award: 1 RO3 MH087444-01

Assay Rationale and Summary: Over one third of the world's population is infected with tuberculosis. In 1993, the World Health Organization declared a global health emergency with the resurgence of tuberculosis in the setting of HIV infection and with the emergence of multi-drug resistant TB. In 2004, 14.6 million people had active TB, 8.9 million new cases were documented, and 1.7 million deaths were attributed to TB (1). Future projections are even more alarming, due to the catastrophic synergy between TB and HIV. In this setting however, we have been forced to rely on suboptimal drugs that were developed in the 1950-60's, with TB drug development vanishing after the 1970's. At present, chemotherapy for TB requires at least six months of a complex multi-drug regimen. Treatment length greatly hinders effective TB control due to the challenges of compliance, which contribute to the development of multi-drug resistance. Despite the alarming implications for public health, research funding has lagged far behind the problem, and the pharmaceutical industry has all but ignored TB (2). New agents that achieve more rapid cures would transform the current pandemic; however, major barriers exist to developing such novel therapeutics including the lack of understanding of the in vivo physiologic states of TB bacilli as they adapt to the host microenvironment in order to survive for extended periods of time.

TB pathogenesis manifests in many forms from active disease to clinical latency that extends for decades requiring long, inefficient antibiotic treatment. The latent form of TB has presented researchers with challenges to understanding this non-replicative, metabolically inactive and drug-tolerant form of TB. Recently hypoxia-induced models that recapitulate the characteristics of the non-replicating, antibiotic resistant state have offered insight into TB lifecycle. Several groups have screened compound libraries with hypoxia induced non-replicative Mycobacterium tuberculosis (M. tb), but to this date, no one has utilized a carbon starvation model to study TB non-replicative dormancy.

Dr. Hung has developed and validated a successful high throughput screen utilizing a carbon-starvation assay using non-replicating, antibiotic-tolerant M. tb to screen small molecule candidates that can be identified as targeting essential functions in M. tb during different physiologic states. For this assay, we usedM. tb H37Rv. Cells are starved for 5 weeks and then plated into 384 well plates where they are exposed to a chemical library for 5 days. Survival of cells is determined by measuring outgrowth in each well after addition of 5x rich media for 4 days. This outgrowth screen identifies several classes of compounds: 1. inhibitors of outgrowth, 2. inhibitors of viability under carbon-starvation, and 3. inhibitors of the transition from carbon-starved to replicating state.

Summary of the Primary Assay
PubChem AID 488890
Test concentration: 25 uM
Activity criterion: >80% inhibition
Number compounds evaluated: 335,740
Number of active compounds: 13,171 of which 2,294 were selected based on the stage they were thought to inhibit, the carbon starved state and the transition between non-replicating and replicating.

Summary of the Confirmatory Assay
PubChem AID 488890
Test concentration: 50-0.1 uM
Activity criterion: IC50 < 10
Number compounds evaluated: 2,294
Number of active compounds: 1,277

Summary HepG2 cytotoxicity of primary screen hits
PubChem AID 489018
Test concentration: 20-0.039 uM
Activity criterion: Minimum Cell Viability < 70%
Number compounds evaluated: 2,294
Number of active compounds: 350

Summary THP-1 cytotoxicity of primary screen hits
PubChem AID 489025
Test concentration: 40-0.078 uM
Activity criterion: Minimum Cell Viability < 70%
Number compounds evaluated: 2,294
Number of active compounds: 789

Summary Vero E6 cytotoxicity of primary screen hits
PubChem AID 492998
Test concentration: 20-0.039 uM
Activity criterion: Minimum Cell Viability < 70%
Number compounds evaluated: 2,294
Number of active compounds: 554
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML338
PubChem Substance ID (SID) for probe 1: 144186624
PubChem Compound ID (CID) for probe 1: 60182306
Probe type for probe 1: Inhibitor
Target for probe 1: Non-replicating M. tuberculosis
Disease relevance for probe 1: Tuberculosis
Anti-target for probe 1: Replicating M. tuberculosis
Fold selectivity for probe 1: >62
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK169453/ (ID: 3060676)
Grant number for probe 1: MH087444-01
NCBI Book chapter title for probe 1: Elucidation of the physiology of non-replicating, drug tolerant Mycobacterium tuberculosis with the aid of the small molecule probe ML338
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
Additional Information
Grant Number: 1 RO3 MH087444-01

Data Table (Concise)
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