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BioAssay: AID 1561

Secondary Assay for lipid storage modulators. Cytotoxcity

NCGC Assay Overview: Storing lipids as a reservoir for energy or the anabolism of elementary metabolites is a common feature of probably all cells and is conserved from bacteria to humans. The universal cellular lipid storage organelle is the so-called lipid storage droplet (LD). Although ubiquitous, LDs share a simple structure composed of a hydrophobic core that harbors the storage lipids, more ..
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 Tested Compounds
 Tested Compounds
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Inactive(1)
 
 
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 Related BioAssays
 Related BioAssays
AID: 1561
Data Source: NCGC (LDSA209)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2009-03-18

Data Table ( Complete ):           All
Tested Compound:
Depositor Specified Assays
AIDNameTypeProbeComment
1519qHTS Assay for Lipid Storage Modulatorsconfirmatory
504433Secondary Assay for lipid storage modulators: Cytotoxcity for probe SAR round 2confirmatory
1623qHTS Assay for Lipid Storage Modulators: Summarysummary1
488908Secondary Assay for lipid storage modulators: Cytotoxcity for probe SARconfirmatory
488900Secondary Assay for lipid storage modulators: Hela cell LDSA assay for probe SARconfirmatory
504432Confirmation Concentration-Response Assay for lipid storage modulators: for probe SAR round 2confirmatory
488913Confirmation Concentration-Response Assay for lipid storage modulators: for probe SARconfirmatory
Description:
NIH Molecular Libraries Probe Production Centers Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
MLPCN Grant: TBA
Assay Submitter (PI): Beller, Mathias; Max-Planck-Institut fur Biophysikalische Chemie, Germany.

NCGC Assay Overview: Storing lipids as a reservoir for energy or the anabolism of elementary metabolites is a common feature of probably all cells and is conserved from bacteria to humans. The universal cellular lipid storage organelle is the so-called lipid storage droplet (LD). Although ubiquitous, LDs share a simple structure composed of a hydrophobic core that harbors the storage lipids, which is shielded by a droplet-specific phospholipid monolayer to which proteins are attached. The current model of LD biogenesis involves an incorporation of the lipid core into the membrane leaflets of the endoplasmic reticulum (ER) followed by a subsequent budding-like maturation of a LD, which ultimately pinches off. Once released, LD volume can increase by localized lipogenesis or fusion of existing droplet. Storage lipids are re-mobilized enzymatically by lipase activity. Lipase regulation in the adipocyte is heavily studied and involves multiple components including catecholamine signaling, the LD-associated proteins Perilipin and comparative gene identification (CGI-58) and at least two lipases named hormone sensitive lipase (HSL) and adipocyte triglyceride lipase (ATGL). LD regulation outside of the adipocyte is poorly understood and only few components are known. However, there is an urgent need to learn more about ectopic fat depots as mislocalized storage of lipids, for example in the liver or muscle, is an eminent health problem associated with insulin resistance or the metabolic syndrome. We developed a laser-scanning cytometer assay to enable 1,536-well screening and combined the results of the small molecules screen with an RNAi database based on lipid storage [1]. The compound was tested here for cytotoxcity using CellTiter-Glo.
Protocol
Assay Protocol Summary: Cytotoxcity assay was performed with embryonic Drosophila S3 cells (Bloomington Drosophila Genomics Resource Center [DGRC]). We dispensed 4 uL of cells at 1.25 x 106 cells/mL into Greiner solid white 1,536-well plates with a bottle-valve solenoid-based dispenser (Aurora) to obtain 5,000 cells/well. A total of 23 nL of compound solution of different concentrations were transferred to the assay plates using a Kalypsys pin tool equipped with a 1,536-pin array containing 10-nL slotted pins (FP1S10, 0.457-mm diameter, 50.8 mm long; V&P Scientific). One microliter of oleic acid (400 uM) was added, and the plate was lidded with stainless steel rubber gasket lined lids containing pinholes. After 24 hrs of incubation at 24 degrees Celsius and 95% humidity, 4 uL of CellTiter-Glo (Promega) was added to the wells. Luminescence was detected on the Perkin Elmer ViewLux.
Comment
Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve, single point activity (at highest concentration only), or inactive. See data field "Curve Description".
2. For all intoxic compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all cytotoxic compounds, a score range was given for each curve class type given above. Cytotoxic compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.0000054935 uM (5.49352e-06μM**)% Activity at given concentration.Float%
15Activity at 0.0000109870 uM (1.0987e-05μM**)% Activity at given concentration.Float%
16Activity at 0.0000219741 uM (2.19741e-05μM**)% Activity at given concentration.Float%
17Activity at 0.0000439481 uM (4.39481e-05μM**)% Activity at given concentration.Float%
18Activity at 0.0000878962 uM (8.78962e-05μM**)% Activity at given concentration.Float%
19Activity at 0.0001757925 uM (0.000175793μM**)% Activity at given concentration.Float%
20Activity at 0.0003515850 uM (0.000351585μM**)% Activity at given concentration.Float%
21Activity at 0.0007031700 uM (0.00070317μM**)% Activity at given concentration.Float%
22Activity at 0.00141 uM (0.00140634μM**)% Activity at given concentration.Float%
23Activity at 0.00281 uM (0.00281268μM**)% Activity at given concentration.Float%
24Activity at 0.00563 uM (0.00562536μM**)% Activity at given concentration.Float%
25Activity at 0.011 uM (0.0112507μM**)% Activity at given concentration.Float%
26Activity at 0.023 uM (0.0225014μM**)% Activity at given concentration.Float%
27Activity at 0.045 uM (0.0450029μM**)% Activity at given concentration.Float%
28Activity at 0.090 uM (0.0900058μM**)% Activity at given concentration.Float%
29Activity at 0.180 uM (0.180012μM**)% Activity at given concentration.Float%
30Activity at 0.360 uM (0.360023μM**)% Activity at given concentration.Float%
31Activity at 0.720 uM (0.720046μM**)% Activity at given concentration.Float%
32Activity at 1.440 uM (1.44009μM**)% Activity at given concentration.Float%
33Activity at 2.880 uM (2.88018μM**)% Activity at given concentration.Float%
34Activity at 5.760 uM (5.76037μM**)% Activity at given concentration.Float%
35Activity at 11.52 uM (11.5207μM**)% Activity at given concentration.Float%
36Activity at 23.04 uM (23.0415μM**)% Activity at given concentration.Float%
37Activity at 46.08 uM (46.0829μM**)% Activity at given concentration.Float%
38Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
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