Inheritedcongenitalanteriorsegmentdysgenesis(ASD)disordersareamajorcauseofvision impairmentandblindness.Theyincludedevelopmentaloculardeficienciesoftheiris,corneaandlens inadditiontosignificantpredisposition,~50%,toglaucoma.Fromvariousstudiesinrecentyearsit hasbecomeclearthatmanycongenitaldisordersandpredispositionsarisefromearlydevelopmental defects,bothmajorandsubtle.Examiningthedevelopmentalprocessgivingrisetotheseessential componentsofthevisualsystemisthereforeafruitfulapproachtostudyingthedisease.Theanterior segmentderiveslargelyfromtheearlymigratingneuralcrestcellsthattargetspecificallytothe anteriorofthedevelopingretinaandarelooselyclassifiedasperiocularmesenchyme(POM).POM deficiencyhasbeendemonstratedincertaincasesofASD.Ibelieveamajorroadblockineffortsto treatandunderstandASDhasbeenthelimitedexaminationofPOMbiologyandfunctionduringearly oculardevelopment.Thishamperseffortstocharacterizetheetiologyofdiseaseandscreeningin ASDfamilies,inadditiontodelayingattemptsatiPSC-mediatedapproachesfortreatmentofASD.To circumventtheseproblems,weaimtocomprehensivelycharacterizePOMcellspecification, migration,targeting,differentiation,andultimatelycontributiontoanteriorsegmentdysgenesis.Thisis thenextlogicalandnecessarystepinorderforthefieldtocontinuetheevolutionoftherapyand screeningforglaucomaandASDdisorders.OurimmediategoalistotestthehypothesisthatPOM cellsdelineateintovarioussubpopulationsandspecifytobecometheanteriorsegment mesenchyme(ASM).Totestourhypothesisweproposetoemploythehighlyversatilezebrafish embryomodelsystemandcomprehensivelyexaminePOMcelldevelopmentwhilecatalogingtheir contributiontotheanteriorsegmentthroughgenetic,molecularandcuttingedgeinvivoimaging approaches.
Our specificaims are:
Aim1 :Characterizespecificationanddefinethecompositionof theASMtheusingacombinationofgeneexpressionanalysispairedwithlight-sheetinvivotime lapseimagingandconfocalmicroscopy.
Aim2 :Trackthemigratorybehaviorandlineagetrace distinctpopulationsofASMduringanteriorsegmentformationusinglight-sheetinvivotimelapse imagingandconfocalmicroscopypairedwithanovelfatemappingoptogenetictracingmechanism.
Aim3 :GenerateacomprehensiveASMtranscriptomeforscreeningnoveltargetsinASDpatient samplesusingacombinationofflowcytometrycoupledtoRNAseqandnextgenerationexome sequencing.

Public Health Relevance

/RELEVANCETOHUMANHEALTH: Anteriorsegmentdysgenesis(ASD)disordershavedevelopmentalunderpinningsthatarecrucialto ourcomprehensionofvision,diseaseandadvancementoftherapy.Clinicalpracticewilldirectly benefitfromstudiesofanimalmodelslikezebrafish,whichprovidestremendousresearch advantagesincludingexuterodevelopment,opticaltransparencythroughoutdevelopmentand capacityforgenomeengineeringanddiseasemodeling.Thegoalofourproposalistogeneratethe firstcomprehensiveclassificationofperiocularmesenchymecellsinordertofacilitateeffortsin screeningforASDpredispositionandenablefutureinvivostemcelltherapy.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
1R01EY027805-01A1
Application #
9448681
Study Section
Biology of the Visual System Study Section (BVS)
Program Officer
Liberman, Ellen S
Project Start
2018-02-01
Project End
2023-01-31
Budget Start
2018-02-01
Budget End
2019-01-31
Support Year
1
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Kentucky
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40526