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BioAssay: AID 651635

qHTS for Inhibitors of ATXN expression

Spinocerebellar ataxia type 2 (SCA2) is a multisystem neurodegenerative disease caused by a dominantly-acting mutation leading to expansion of a polyQ domain in the ataxin-2 protein (ATXN2). SCA2 patients develop progressive ataxia and later lose function in other neuronal systems. Prominent parkinsonian signs develop in some patients. Similar to many neurodegenerative disorders, no symptomatic or disease-modifying treatments are known. ..more
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 Tested Compounds
 Tested Compounds
All(391566)
 
 
Active(3812)
 
 
Inactive(343445)
 
 
Inconclusive(44879)
 
 
 Tested Substances
 Tested Substances
All(394495)
 
 
Active(3833)
 
 
Inactive(345643)
 
 
Inconclusive(45019)
 
 
AID: 651635
Data Source: NCGC (SCA2100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2012-10-11
Modify Date: 2012-10-18

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 3812
Depositor Specified Assays
AIDNameTypeComment
588380qHTS for Inhibitors of ATXN expression: SummarysummarySummary AID
Description:
Spinocerebellar ataxia type 2 (SCA2) is a multisystem neurodegenerative disease caused by a dominantly-acting mutation leading to expansion of a polyQ domain in the ataxin-2 protein (ATXN2). SCA2 patients develop progressive ataxia and later lose function in other neuronal systems. Prominent parkinsonian signs develop in some patients. Similar to many neurodegenerative disorders, no symptomatic or disease-modifying treatments are known.

The primary objective of the proposed research is to identify compounds inhibiting ATXN2 expression and to test them for efficacy in SCA2 mouse models. To this end, we have developed a cell-based assay paired with an in vivo mouse model using identical luciferase expression constructs. This approach takes advantage of an ATXN2-luciferase transgene, with the luciferase gene flanked by the upstream and downstream portions of the ATXN2 gene. This primary assay was tested against the NIH Molecular Libraries Small Molecule Repository (MLSMR).

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: NS073009
Assay Submitter (PI): Daniel Scoles, University of Utah
Protocol
4 microl of SH-SY5Y-ATXN2-FFLuc cell suspension (TBD cells/microL in phenol-red free DMEM) are dispensed into Greiner white solid bottom 1536-well assay plates, yielding a final well density of TBD cells/well. Plates are incubated for overnight at 37 degrees C in 5% humidified CO2 to allow cell attachment. Compounds are then transferred via Kalypsys pin tool equipped with 1536-pin array (10nl slotted pins, V&P Scientific, San Diego, CA). After incubation at 37 degrees C in 5% humidified CO2 for 24 hrs, 1microl of GF-AFC (Gly-Phe-7-amino-4-trifluoromethyl coumarin, MP Biomedicals) at a final concentration of 25microM is then added and plates are incubated for 30 minutes, before being transferred to a ViewLux high-throughput CCD imager (PerkinElmer), wherein single end-point fluorescence measurements are acquired using the 405 nm excitation/525 nm emission filter set to assess cell viability. Next, Steady-Glo luciferase substrate detection reagent (2microl) (Promega, Madison, WI) is added to each well and incubated for an additional 5 minutes. Luminescence is then measured on the ViewLux imager equipped with a clear emission filter using a 2 sec exposure.
Comment
Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active 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
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
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
5Activity_ScoreActivity score.Integer
6Curve_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
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_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
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.00366 uM (0.00366μM**)% Activity at given concentration.Float%
16Activity at 0.042 uM (0.0424412μM**)% Activity at given concentration.Float%
17Activity at 0.091 uM (0.0914μM**)% Activity at given concentration.Float%
18Activity at 0.191 uM (0.190526μM**)% Activity at given concentration.Float%
19Activity at 0.229 uM (0.229407μM**)% Activity at given concentration.Float%
20Activity at 0.457 uM (0.456731μM**)% Activity at given concentration.Float%
21Activity at 0.981 uM (0.981014μM**)% Activity at given concentration.Float%
22Activity at 1.459 uM (1.45901μM**)% Activity at given concentration.Float%
23Activity at 2.289 uM (2.28856μM**)% Activity at given concentration.Float%
24Activity at 4.540 uM (4.5404μM**)% Activity at given concentration.Float%
25Activity at 5.375 uM (5.37544μM**)% Activity at given concentration.Float%
26Activity at 11.39 uM (11.3937μM**)% Activity at given concentration.Float%
27Activity at 24.53 uM (24.5317μM**)% Activity at given concentration.Float%
28Activity at 36.51 uM (36.5111μM**)% Activity at given concentration.Float%
29Activity at 57.07 uM (57.0668μM**)% Activity at given concentration.Float%
30Activity at 112.8 uM (112.847μM**)% Activity at given concentration.Float%
31Activity at 131.6 uM (131.583μM**)% Activity at given concentration.Float%
32Activity at 224.9 uM (224.947μM**)% Activity at given concentration.Float%
33Activity at 287.0 uM (287μM**)% Activity at given concentration.Float%
34Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: NS073009

Data Table (Concise)
Classification
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