Bookmark and Share
BioAssay: AID 1827

Broad Institute MLPCN Alpha-Synuclein 5'UTR - Inhibitors

The goal of this project is to identify novel small molecule probes that inhibit alpha-synuclein translational expression in dopaminergic neurons by targeting the 5'-untranslated region (5'UTR) stem-loop of alpha-synuclein as a major new therapeutic target to retard the progression of Parkinson's disease (PD). The 5'UTR of alpha-synuclein mRNA can interact with Iron Regulatory Protein-1 (IRP1), more ..
_
   
 Tested Compounds
 Tested Compounds
All(2)
 
 
Probe(2)
 
 
Active(2)
 
 
 Tested Substances
 Tested Substances
All(2)
 
 
Probe(2)
 
 
Active(2)
 
 
 Related BioAssays
 Related BioAssays
AID: 1827
Data Source: Broad Institute (2023-inhibitors)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2009-06-23
Modify Date: 2011-03-04

Data Table ( Complete ):           Active    All
BioActive Compounds: Chemical Probe: 2    Active: 2
Depositor Specified Assays
Show more
AIDNameTypeComment
1813MLPCN Alpha-Synuclein 5'UTR - 5'-UTR binding - inhibitorsscreeningprimary screening campaign
1988Luminescence Cell-Based Dose Confimation HTS to Identify Inhibitors of of 5'UTR Stem-Loop Driven Alpha-Synuclein mRNA Translation in H4 Neuroglioblastoma CellsconfirmatoryPrimary Dose Retest
2454Luminescence Cell-Based HTS Dose Confimation to Identify Inhibitors of of 5'UTR Stem-Loop Driven Alpha-Synuclein mRNA Translation in H4 Neuroglioblastoma CellsconfirmatoryPrimary Dose Retest
2460Luminescence Cell-Based Dose Confimation HTS to Identify Inhibitors of of 5'UTR Stem-Loop Driven Alpha-Synuclein mRNA Translation in H4 Neuroglioblastoma Cells.confirmatory
2452Luminescence Cell-Based Dose Response to Identify Inhibitors of 5'UTR Stem-Loop Driven Prion Protein mRNA Translation in H4-C Neuroglioblastoma CellsconfirmatoryCounter screen
2458Luminescence Cell-Based Dose Response to Identify Inhibitors of Luciferase Translation or Activity in H4-C Neuroglioblastoma CellsconfirmatorySecondary Screen
2463Luminescence Cell-Based Dose Response HTS to Identify Inhibitors of Luciferase Translation or Activity in H4-C Neuroglioblastoma Cellsconfirmatory
2468Luminescence Cell-Based Dose HTS Response to Identify Inhibitors of 5'UTR Stem-Loop Driven Prion Protein mRNA Translation in H4-C Neuroglioblastoma Cellsconfirmatory
2471Luminescence Cell-Based Dose-Confimation HTS to Identify Inhibitors of of 5'UTR Stem-Loop Driven Alpha-Synuclein mRNA Translation in H4 Neuroglioblastoma Cellsconfirmatory
2484Western Blot Cell-Based Dose Response to Identify Inhibitors of Binding of Alpha-Synuclein Translation in H4 Cellsconfirmatory
2473ELISA Cell-Based Dose Response to Identify Inhibitors of Alpha-Synuclein Translation in SH-SY5Y Cellsconfirmatory
1990Luminescence Cell-Based Dose Response HTS to Identify Inhibitors of Luciferase Translation or Activity in H4 Neuroglioblastoma CellsconfirmatorySecondary Screen
1994Luminescence Cell-Based Dose Response HTS to Identify Inhibitors of 5'UTR Stem-Loop Driven Prion Protein mRNA Translation in H4 Neuroglioblastoma CellsconfirmatoryCounter Screen
Description:
Broad Institute of MIT and Harvard, Cambridge MA

NIH Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Number 1-R21-NS-059434-01

Internal Project ID: 2023

Keywords: alpha-synuclein, Parkinson's disease, translation, RNA stem-loop, 5'-untranslated region

Primary Collaborators:

Jack T. Rogers, Massachusetts General Hospital, Charlestown, MA, jtrogers@rics.bwh.harvard.edu

Probes:
ML150

Project Overview:

The goal of this project is to identify novel small molecule probes that inhibit alpha-synuclein translational expression in dopaminergic neurons by targeting the 5'-untranslated region (5'UTR) stem-loop of alpha-synuclein as a major new therapeutic target to retard the progression of Parkinson's disease (PD). The 5'UTR of alpha-synuclein mRNA can interact with Iron Regulatory Protein-1 (IRP1), which upon interaction causes an increase in alpha-synuclein mRNA translation. Probes that can successfully reduce alpha-synuclein expression levels as measured in this luciferase reporter assay will be further tested for specificity in cells lacking the alpha-synuclein 5'UTR stem-loop to confirm that probes are acting through the intended target. Probe selectivity will be tested in cells containing the prion protein 5'UTR mRNA stem-loop fused to a luciferase reporter.

Primary Assay Summary:

Name: 5'-UTR Binding

Internal Assay ID: 2023-01

Assay Overview:

Compounds were assayed against H4 neuroblastoma cells transfected with a plasmid containing alpha-synuclein 5'UTR-luciferase gene fusion (Jack Rogers). Compounds that cause a reduction in luciferase expression will be further tested for proper target binding and specificity. Assays were conducted in 384-well format (Corning, 3570) with a 3000 cells/well coating density in 50 uL phenol red-free DMEM with 4.5 g/L D-glucose (Lonza, 12-917F) supplemented with 10 % FBS (Lonza, 14-503E), 200 uM L-glutamine (Lonza, 17-605E), 100 uM penstrep (Lonza, 17-602E) and 200 ug/mL geneticin (Invitrogen, 10131-027). Following overnight growth, 100 nL of 3.75 mM compound was added to the cell solutions (7.5 uM final [compound]). The positive control used in this assay was strophanthidine (Sigma, Cat.# S6626-250MG, Lot#038K1036), which was used in dose (12-point, 2-fold dilution, from 20 uM final [compound]) and was incorporated at the beginning and end of each days run. Cells were grown for 48 h, equilibrated to room temperature and 30 uL Steady-Glo (Promega, E2250) reagent was added. The plates were then incubated at room temperature for 30 min, and luciferase levels were measured.

PubChem AID 1813

Number substances evaluated: 303857

Since the calibration control used acts as an inhibitor of this assay, activators were identified by selecting compounds with a negative PubChem_Activity_Score.

Active Compounds : Activity_Score >= 62.
62 represents a score that is 4x the standard deviation of all negative control (DMSO) samples in the screen.

Number of active compounds: 2498

Inconclusive Compounds: Activity_Score < 62 with at least one replicate activity score >=62.

Number of inconclusive compounds: 481
Comment
The following compounds [SID] were purchased or synthesized for this project:
87218804 87218892 87218893 87218896 87218898 87218900
87218901 87218902 87218903 87218905 87218906 87218907
87218908 87218909 87218910 87457149 87457152 87556803
87556804 87556806 87556807 87556808 87556809
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1ML_NUMML number assigned to compoundString
Additional Information
Grant Number: 1-R21-NS-059434-01

Data Table (Concise)
PageFrom: