Abstract

Drugdiscoverypipelinesforneuropsychiatricdisordersaredry.Oneapproachtorejuvenatingthese pipelineswouldbetocreateassaysbasedonrelevantdiseasephenotypesinprimaryneurons,somethingthat iscurrentlylacking.However,ascalableassaydevelopmentplatformthatisbasedonbonafideneurons, remainscosteffective,andthatcansupportindustriallevelhigh-throughputscreening(HTS)doesnotcurrently exist.Overthepasteightyears(spreadacrossdifferentNIH-sponsoredgrants),ourcollaborativegrouphas createdaflexibleandscalableprimaryneuron-basedassaydevelopmentsystemthatiscompatiblewith industrial-levelHTS.Ourlong-standinggoalforthisprojecthasbeentooptimizetheseproceduresand workflowstosupportneuron-basedHTSphenotypicassayssothattheycansupportverylarge screeningcampaignsofupto200Kcompounds. Wearehappytoreportthatprogressoverthelastbudgetperiodhaspushedusclosertowardthis statedgoal.Wehaveinventedastate-of-the-art,disease-modelingassaycreatedinprimaryneuronsthatis designedtodiscovercompoundsthatreversethecellularconsequencesofgenetichaploinsufficiency.Indeed, asubstantialproportionofchildhoodbraindisordersarecausedbysingleautosomaldominantvariants resultingingenetichaploinsufficiency.Theraregeneticbraindisordersthatarisefromthesevariantsoffer greatpotentialfortranslationbecausethediseasemechanismiswell-understood(i.e.lowproteinexpression). Therefore,arationaleprecisiontherapyfortreatinggenetichaploinsufficiencydisorderswouldbetodiscover ?magicbullet?compoundsthatraiseexpressionoffunctionalproteinsfromtheremainingundamagedallele (e.g.?boostingcompounds?).Inthisrenewalproject,wewillemploytechnicalinnovationsthathaveunlocked thescalabilityofprimaryneuronsforphenotypicHTS.Asaproof-of-principle,wewillscale-upand implementanassaythatreportsreversaloflowSynGAPexpressioninneuronscausedbygenetic haploinsufficiencyoftheSYNGAP1/Syngap1gene.WewillminiaturizeanHTS-compatibleanddisease- modelingsteady-stateendogenousSynGAPexpressionassaysothatitiscompatiblewithindustrialscaleHTS automation.Onceimplemented,wewillthenscreenupto200,000uniquesubstancesusingacompletely automatedversionoftheneuron-basedSynGAPexpressionassay.Finally,usingacomprehensivemulti-stage biologicalvalidationfunnel,wewillidentifyandprioritizethemosttranslatablechemicalprobesthatraise SynGAPproteinexpression.Theoverallimpactofthisprojectisthatdiscoveryofmultiple,validatedSynGAP boostingcompoundswouldprovideproof-of-principlethatourflexibleplatformisaneffectivetoolfor phenotypicdrugdiscoveryfornervoussystemdisorders.

Public Health Relevance

OneapproachtorejuvenatingdrugpipelinesforneuropsychiatricdisordersistocreateHTSscalableassays basedonrelevantdiseasephenotypesinprimaryneurons,thoughacosteffectiveplatformfortrueHTS- compatibleneuron-basedphenotypicassaysdoesnotcurrentlyexistinacademia.Asameanstorefineour novelneuron-basedHTSsystem,andtoadvancetreatmentforASD-relateddisorders,weareseekingto scale-upandimplementaprimaryneuron-basedphenotypicassaythatcanrevealchemicalprobescapableof reversingtherootcauseofararegeneticbraindisorder.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
2R01MH113648-04A1
Application #
10063708
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Michelotti, Enrique
Project Start
2017-05-01
Project End
2024-02-29
Budget Start
2020-05-01
Budget End
2021-02-28
Support Year
4
Fiscal Year
2020
Total Cost
Indirect Cost

  Institution

Name
Scripps Florida
Department
Type
DUNS #
148230662
City
Jupiter
State
FL
Country
United States
Zip Code
33458

  Publications

Spicer, Timothy P; Hubbs, Christopher; Vaissiere, Thomas et al. (2018) Improved Scalability of Neuron-Based Phenotypic Screening Assays for Therapeutic Discovery in Neuropsychiatric Disorders. Mol Neuropsychiatry 3:141-150
Kilinc, Murat; Creson, Thomas; Rojas, Camilo et al. (2018) Species-conserved SYNGAP1 phenotypes associated with neurodevelopmental disorders. Mol Cell Neurosci 91:140-150