Abstract

Autosomaldominantmutationsinvacuolarproteinsorting35(VPS35)havebeenidentifiedasacausalPD gene,playingaroleinthedevelopmentoflate-onsetPD.VPS35functionsasascaffoldingproteinforretromer complexthatmediatesrecyclingofcargoproteins(transmembraneproteins)fromendosomestothetrans- Golgiapparatusortheplasmamembrane.Therefore,mutationsinVPS35couldpreventorlimititsdeliveryof thecargoes,whicharecrucialforthesurvivalofdopaminergicneurons.Hence,adisruptionintherecycling pathwayofthesecargoesmayplayacrucialroleinthedemiseofdopaminergicneuronsinthesubstantia nigraofpatientswithVPS35mutations.AgreaterunderstandingofbiologyofVPS35andthepathophysiology ofmutantVPS35areessentialtothedevelopmentoftherapeuticinterventionsaimedatpreventingtheonset and/orretardingprogressionofPD.Nonetheless,thefunctionalconsequencesoftheVPS35geneticvariations inPDhavenotyetbeendiscovered.TobetterunderstandthepathogenicinvolvementofVPS35mutationsin vivo,wegeneratedatetracyclineconditionalhumanVPS35transgenic(Tg)mousewhereexpressionof mutanthumanD620NVSP35orwild-type(WT)humanVPS35proteinsisachievedinthenigrostriatal dopaminergicpathway,underthecontrolofthedopaminepathway-specifictyrosinehydroxylase(TH)-tTA driver.Utilizingthismousemodel,inaim1,wewillstudyneurochemical,neuroanatomicalandbehavioral changesusinghigh-performanceliquidchromatography,unbiasedstereologicaltechniques,andbehavioral testinginthesemiceastheyage.Inparticular,wewillexplorewhethertheoverexpressionofmutantD620N VPS35indopaminergic(DA)neuronsmayinducelossofdopaminergicneuronsduringaging.Importantly,our preliminarystudyindicatesthattherearerobustandprogressivedegenerationinthesubstantianigraofthe TH-tTA/TetP-D620NVPS35mice.Inaddition,thereisanintriguingbutpoorunderstandingofthepathogenic interplaybetweentheVPS35mutationandmitochondriadysfunctioninPD.Ourpreliminaryresultindicates thatVPS35interactswithKeap1andtheinteractionbetweenKeap1andD620NVPS35leadstoaccumulation ofKeap1,akeyregulatorofNrf2,andaconcomitantdecreaseinproteinlevelsandactivityofNrf2,amaster regulatorofoxidativestress.Inaddition,thedysregulationofKeap1/Nrf2levelsmediateD620NVPS35- inducedDAneuronaltoxicityandmitochondriadysfunctioninhumanDAneurons.
In aim2, wewill characterizeapotentialdefectinmitochondrialqualitycontrolintheD620NVPS35Tgmicerepresenting degenerationofDAneuronsandtheroleofthederegulationofKeap1/Nrf2levelsinregulatingthesedefects. Moreover,wewilldeterminewhethersuppressionofKeap1accumulationrescuesthelossofDAneuronsand mitochondriadysfunctionsinD620NVPS35Tgmice.Thisproposalmayprovideaneworvaluablegenetic mousemodelforPDwithdopaminergicneurodegenerationandanewinsightofVPS35retromerfunctionin lossofdopaminergicneuronsandmitochondriadysfunction.

Public Health Relevance

(RELEVANCE) MutationsinVPS35arethecommoncauseoffamilialPD,yetlittleisknownabouthowthisproteinfunctionsin thepathogenesisofPD.Decipheringofalteredretromerfunctionindegenerationofdopaminergicneurons throughgeneratingandcharacterizingWTandD620NVPS35transgenicmiceandunderstandingapotential pathogenicinterplaybetweenVPS35mutationandmitochondrialdysfunctionswillfurtherenhanceour understandingofVPS35-linkedPDandpromotedevelopmentofdrugsthatsloworhaltthedisease.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21NS098006-01A1
Application #
9316771
Study Section
Cell Death in Neurodegeneration Study Section (CDIN)
Program Officer
Sieber, Beth-Anne
Project Start
2017-02-01
Project End
2019-01-31
Budget Start
2017-02-01
Budget End
2018-01-31
Support Year
1
Fiscal Year
2017
Total Cost
$244,938
Indirect Cost
$94,938

  Institution

Name
Johns Hopkins University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21205

  Publications

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Yun, Seung Pil; Kam, Tae-In; Panicker, Nikhil et al. (2018) Block of A1 astrocyte conversion by microglia is neuroprotective in models of Parkinson's disease. Nat Med 24:931-938
Kim, Donghoon; Hwang, Heehong; Choi, Seulah et al. (2018) D409H GBA1 mutation accelerates the progression of pathology in A53T ?-synuclein transgenic mouse model. Acta Neuropathol Commun 6:32
Kim, Sangjune; Yun, Seung Pil; Lee, Saebom et al. (2018) GBA1 deficiency negatively affects physiological ?-synuclein tetramers and related multimers. Proc Natl Acad Sci U S A 115:798-803
Yun, Seung Pil; Kim, Donghoon; Kim, Sangjune et al. (2018) ?-Synuclein accumulation and GBA deficiency due to L444P GBA mutation contributes to MPTP-induced parkinsonism. Mol Neurodegener 13:1
Dawson, Ted M; Golde, Todd E; Lagier-Tourenne, Clotilde (2018) Animal models of neurodegenerative diseases. Nat Neurosci 21:1370-1379
Lee, Saebom; Kim, Sangjune; Park, Yong Joo et al. (2018) The c-Abl inhibitor, Radotinib HCl, is neuroprotective in a preclinical Parkinson's disease mouse model. Hum Mol Genet 27:2344-2356
Yun, Seung Pil; Kim, Hyojung; Ham, Sangwoo et al. (2017) VPS35 regulates parkin substrate AIMP2 toxicity by facilitating lysosomal clearance of AIMP2. Cell Death Dis 8:e2741
Brahmachari, Saurav; Karuppagounder, Senthilkumar S; Ge, Preston et al. (2017) c-Abl and Parkinson's Disease: Mechanisms and Therapeutic Potential. J Parkinsons Dis 7:589-601