Thereisacriticalneedforimprovedhumantissuemodelstostudyinfectiousdiseases.Animalmodelsoften failtoreproducehumanphysiology,andaresimilarlypoorpredictorsofdrugefficacywhentranslatedto humans.ThisproposaldescribestheMITCenterforHumanTissueModelsforInfectiousDiseases (MIT.HTMID),whichwillfocusontwodimensionalhumanneuralcellsandthreedimensionalhumancerebral organoidstostudyvirusinfections.TheprojectissitedentirelyatTheMassachusettsInstituteofTechnology. Thethreeinvestigatorsare:LeeGehrke(VirologyandInfectiousDiseases),RudolfJaenisch(Tissue Engineering,CellBiology,andStemCells)andDavidSabatini(GeneticsandScreeningTechnologies).The CenterwillalsoincludethreeCores?thatis,Administrative,Virology,andHumanCellsandTissues.The themesofthetwointerrelatedResearchProjectsofthisU19proposalare:?Project1:?Humantissuemodelsto studyinfectiousdiseases:Human2Dand3Dneuralculturesforstudyingvirustropismandinfection phenotypes,and?Project2:?Useof2Dculturesand3Dorganoidstoidentifycandidateantiviralcompounds?to usegeneticapproachestoidentifyandvalidatehostgenesthatpromoteorprotectagainstflavivirusinfection?. Theexperimentaluseoforganoidsissignificantbecausethethreedimensionalarchitectureanddifferentiation fromembryonicstem(ES)cellsandinducedpluripotentstem(iPS)cellsprovidenearphysiologicalfunctionsin tissueorganization,tissuerenewal,andresponsestopathogeninfections.Indeed,humanorganoidshave beengeneratedforawiderangeoftissuesandusesinstudyingdevelopmentanddiseases,includingvirus infections.Theprojectwillcomparetheinfectionsoffivedifferentneuralormicroglialcelltypes(neuronal progenitors,neurons,oligodendrocytes,astrocytes,microglia)withthreeflaviviruses(ZikaVirus,WestNile Virus,orDenguevirus).TheresearchgoalsofMIT.HTMIDaddresstheZikavirusglobalhealthcrisis,toward understandinghowrelatedflavivirusescancauseverydifferentdiseases,includingmicrocephalyandGuillain BarreSyndrome.Thevirusworkwillbeextendedbeyondflavivirusestoincludeotherneurotropicviruses?that is,pseudotypedvesicularstomatitisviruses(VSV)thatcarrytheenvelopesofselectagentencephaliticviruses (EasternEquineEncephalitis,WesternEquineEncephalitis,andVenezuelanEquineEncephalitis).Wewill usecellandmolecularmethodstodefineandcomparetheinfectionphenotypesofthecellsandviruses. TissueengineeringandgeneticswillbecombinedbyperformingCRISPRCas9screenstoidentifygenesthat regulateorareregulatedbyvirusinfection,andthengenerating?knockout?organoidstotestfunctionina threedimensionaltissue.Theorganoidhumantissuemodelwillalsobeusedtoevaluateanumberofantiviral compoundstovalidateitspotentialuseasadrugtestingplatform.Ifsuccessful,thisCenterwillyieldsignificant newdataandbeanintegralcomponentofanetworkofHumanTissueModelsforInfectiousDiseases.
This proposal describes the MIT Center for Human Tissue Models of Infectious Diseases (MIT.HTMID), which will study viral infections of biologically relevant human neural cells and cerebral organoids that are differentiated from stem cells. The proposal includes three investigators, representing expertise in stem cells and tissue engineering, virology, and human genetics. If successful, the MIT.HTMID center will develop and apply new human tissue models to investigate infectious diseases, with emphasis on understanding Zika virus microcephaly.
