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BioAssay: AID 588528

Broad Institute Reversing Antifungal Drug Resistance: Single Agent Probe Inhibitor Probe Project

to delineate the metabolic requirements for fungal virulence. Probes will be evaluated in genetic studies using genome-wide over-expression and deletion libraries to define their modes of action in S. cerevisiae assayed under fermentative versus respiratory growth conditions. To complement genetic approaches, affinity precipitation and proteomic approaches based on Stable Isotope Labeling with more ..
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 Related BioAssays
 Related BioAssays
AID: 588528
Data Source: Broad Institute (2148_Inhibitor_Project)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2011-10-17
Modify Date: 2012-04-11
Depositor Specified Assays
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AIDNameTypeComment
588530CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_CherryPick_ActivityconfirmatoryCaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader
588529Anti-Fungal Single Agent: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_CherryPick_ActivityconfirmatoryAnti-Fungal Single Agent: CaCi2 Measured in Microorganism System Using Plate Reader
1979Fluorescence Cell-Based Primary HTS of C.albicans growth in the presence of Fluconazole and compoundscreening302509 hts compounds at singlepoint in 01 Antifungal Primary measuring activity
588634Antifungal Single Agent - Mammalian Cell Toxicity Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2148-03_Inhibitor_Dose_CherryPick_Activityconfirmatory39 cherrypick compounds at dose in Secondary-3T3 Fibroblasts measuring activity
602190Anti-Fungal Single Agen: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose in CaCi2 Primary measuring activity
602189CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose in CaCi17 Primary measuring activity
602188Antifungal Single Agent - Mammalian Cell Toxicity Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2148-03_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose Secondary in 3T3 Fibroblasts measuring activity
602186Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose in C. glabrata glycerol measuring activity
602185Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose in Glabrata Glucose measuring activity
602184Anti-Fungal Single Agent: S.cerevisiae growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-06_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose in S cerevisiae glycerol measuring activity
602187Anti-Fungal Single Agent: S.cerevisiae growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-07_Inhibitor_Dose_DryPowder_Activityconfirmatory24 drypowder compounds at dose in S cerevisiae glucose measuring activity
602394Antifungal Single Agent - Mammalian Cell Toxicity Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2148-03_Inhibitor_Dose_DryPowder_Activity_Set2confirmatory55 drypowder compounds at dose in Secondary 3T3 Fibroblasts measuring activity set 2
623904Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activity_Set2confirmatory55 drypowder compounds at dose in Glabrata Glucose measuring activity set 2
623899Anti-Fungal Single Agent: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activity_Set2confirmatory55 drypowder compounds at dose in CaCi2 Primary measuring activity set 2
623897CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_DryPowder_Activity_Set2confirmatory55 drypowder compounds at dose in CaCi17 Primary measuring activity set 2
623903Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activity_Set2confirmatory55 drypowder compounds at dose in C. glabrata glycerol measuring activity set 2
624006Anti-Fungal Single Agen: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activity_Set4confirmatory9 drypowder compounds at dose in CaCi2 Primary measuring activity set 4
623965Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activity_Set3confirmatory18 drypowder compounds at dose in C. glabrata glycerol measuring activity set 3
623967Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activity_Set3confirmatory18 drypowder compounds at dose in Glabrata Glucose measuring activity set 3
623969Anti-Fungal Single Agent: S.cerevisiae growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-06_Inhibitor_Dose_DryPowder_Activity_Set3confirmatory18 drypowder compounds at dose in S cerevisiae glycerol measuring activity set 3
623971Anti-Fungal Single Agent: S.cerevisiae growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-07_Inhibitor_Dose_DryPowder_Activity_Set3confirmatory18 drypowder compounds at dose in S cerevisiae glucose measuring activity set 3
623981Anti-Fungal Single Agen: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activity_Set3confirmatory9 drypowder compounds at dose in CaCi2 Primary measuring activity set 3
623982CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_DryPowder_Activity_Set3confirmatory9 drypowder compounds at dose in CaCi17 Primary measuring activity set 3
624014Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activity_Set4confirmatory9 drypowder compounds at dose in C. glabrata glycerol measuring activity set 4
624013Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activity_Set4confirmatory9 drypowder compounds at dose in Glabrata Glucose measuring activity set 4
624012Anti-Fungal Single Agent: S.cerevisiae growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-06_Inhibitor_Dose_DryPowder_Activity_Set4confirmatory9 drypowder compounds at dose in S cerevisiae glycerol measuring activity set 4
624025Anti-Fungal Single Agent: S.cerevisiae growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-07_Inhibitor_Dose_DryPowder_Activity_Set4confirmatory9 drypowder compounds at dose in S cerevisiae glucose measuring activity set 4
624068Disruption of Mitochondrial Function in C. glabrata Measured in Microorganism System Using Flow Cytometry - 2148-09_Inhibitor_SinglePoint_DryPowder_Activityother
624062Disruption of mitochondrial function in S. cerevisiae Measured in Microorganism System Using Flow Cytometry - 2148-08_Inhibitor_SinglePoint_DryPowder_Activityother
Description:
Primary Collaborators:
Susan Lindquist,Whitehead Inst.,Cambridge, MA,sll@wi.mit.edu
Luke Whitesell,Whitehead Inst.,sll@wi.mit.edu
Ben Vincent,Whitehead Inst.,sll@wi.mit.edu

Project Goal:

Probes arising from this new project will be useful tools
to delineate the metabolic requirements for fungal virulence. Probes will be evaluated in genetic studies using genome-wide over-expression and deletion libraries to define their modes of action in S. cerevisiae assayed under fermentative versus respiratory growth conditions. To complement genetic approaches, affinity precipitation and proteomic approaches based on Stable Isotope Labeling with Amino acids in Cell culture (SILAC) technology may also be used for target identification. Further development of these probes, including optimization of DMPK properties in animals will be pursued to ask whether mitochondrial function is required for infection by drug resistant C. glabrata and C. albicans in well-established mouse models of human disease.

Probe Attributes:
a. IC50 <10uM for growth inhibition of C. albicans clinical isolate CaCi-2 as single agent.
b. At least 10-fold selectivity between the primary Candida test strain and mammalian cells.
c. Activity against drug-pump overexpressing C. albicans isolate CaCi-17 at an IC50 < 10uM.
d. At least 10-fold selectivity in limiting the growth of S. cerevisiae when cultured in medium
supplemented with glycerol compared to glucose as carbon energy source.
e. At least 10-fold selectivity in limiting the growth of C. glabrata when cultured in medium
supplemented with glycerol compared to glucose as carbon energy source.


Candida albicans, glycolysis, respiration
Biological Relevance:
Intrinsic and acquired drug resistance by medically relevant microorganisms poses a grave threat to human health and has enormous economic consequences. Fungal pathogens present a particular challenge because they are eukaryotes and share many of the same mechanisms that support the growth and survival of the human host cells they infect. The number of drug classes that have distinct targets in fungi is very limited and the usefulness of current antifungal drugs is compromised by either dose-limiting host toxicity or the frequent emergence of high-grade resistance. New, non-cross-reactive targets for therapeutic intervention are urgently needed.

A frequently overlooked issue in the search for new antifungal targets is that fungal pathogens face a diverse array of environmental challenges during the establishment of invasive infection in a host animal. These can include pH, thermal and osmotic stresses as well as the need to utilize alternative carbon sources for energy production. Extrapolating drug sensitivity patterns defined using standard in vitro culture conditions to the setting of host infection can be misleading. A particularly vivid example of this problem has been provided by the demonstration of differential requirements for growth in vitro versus virulence in mice using a library of deletion mutants constructed in a clinical isolate of the yeast S. cerevisiae. The role of glycolytic versus respiratory metabolism in supporting fungal virulence and drug-resistance has been particularly controversial, due in large part to the absence of good tools with which to tackle the question. To address this unmet need, the goal of this project is identify drug-like molecules that selectively inhibit the growth of fungi when they are cultured under conditions that require mitochondrial respiration to support metabolic needs. Given the relatively limited genetic tools available, such compounds will be of great value in probing metabolic requirements for fungal virulence. They could also provide essential leads for the development of new antifungal drugs that operate in a completely unexploited target space.
Additional Information
Grant Number: 1 R03 MH086456-01

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