Abstract

?PROJECT2 Manyunknownpathogenicstepsliebetweennormaltauproteininahealthyneuronandthecellular dysfunctionthatoccursasaresultoftauuptakeandaggregation.
The aims addresstwoemergingfacetsof thepathobiology:themechanismoftauuptakeinneuronsandastrocytesandtheeffectsoftauinclusionsona setoffunctionalcellularparameters.Tauuptakewillbestudiedusingahighlynovelandhighlycollaborative approachcalledCRISPRitoidentifyinanonbiasedmannergenesinvolvedintauuptakefollowedby functionalstudies,whichwillvalidatetheCRISPRihits.Amorefocusedapproachtouptakewillfocuson HSPGswithaGAGmicroarraypaneltoidentifyHSPGchainfeaturestowhichtaucanbindspecifically.Once tauisinsidethecellweaskwhataretheeffectsoftauinclusionsortaumutationsinhumaniPSCderived neurons.Wehavesynthesizedmultipletypesoftauaggregatesandstudiedtheiruptakeconditions.Avariety oftechniqueswillbeappliedtopinpointcellulardefectsduetotheburdenoftauinclusions.Thesetechniques fallintotwocategories?expressionsequencingandelectrophysiologicalassessment.Withregardtothe former,thestrengthofourproposalistheuseofsinglecellRNAseqtoidentifypreciselygeneexpression changesincellswithtauinclusionscomparedtoitsneighborsthatdonothavetauinclusions.Afurther strengthisthedatathatsupportsourhypothesisconcerningtheroleoftRNAintriggeringtauconformational changesthatmayleadtoaggregationaswellasaninvestigationoftheroleofcellularstressininducingtRNA cleavageandtheformationofhalftRNAsknownastiRNAs.Wehavealsohypothesizedthattaucaninduce electrophysiologicdysfunctionandoverthepastthreeyearsincollaborationwiththephysicsdepartmenthave builttoolscapableofdetectingconductiondeficits,alterationsintheactionpotentialattheaxonalinitial segmentandhyerexcitability.Thedetectionmethodsutilizeamultielectrodearray(MEA)platform,modified MEAsthatarepatternedtoconfineneurongrowthanddirectsignalingbetweenneuronalensembles,analytical toolsforspiketrains,andanautomatedcellharvestingdevice.Thedelineationoftheeffectsontaumutations andtauinclusionsonculturedcellsisacellularphenotypethatisnotwelldescribedinthefield,butwillbe necessaryinthefutureforsmallmoleculescreens.Workinginterchangeablywithseveralcellularsystemswill allowustoselecttheidealcontextforeachexperimentalquestionposedandexploremultiplefacetsina searchfortaurelatedcellularphenotypes.Thisproposalrestsonstronginteractionsamongalltheteam members.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54NS100717-04
Application #
9791006
Study Section
Special Emphasis Panel (ZNS1)
Project Start
Project End
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost

  Institution

Name
Weill Medical College of Cornell University
Department
Type
DUNS #
060217502
City
New York
State
NY
Country
United States
Zip Code
10065

  Publications

Tracy, Tara E; Gan, Li (2018) Tau-mediated synaptic and neuronal dysfunction in neurodegenerative disease. Curr Opin Neurobiol 51:134-138
Wang, Chao; Telpoukhovskaia, Maria A; Bahr, Ben A et al. (2018) Endo-lysosomal dysfunction: a converging mechanism in neurodegenerative diseases. Curr Opin Neurobiol 48:52-58
Paulo, Esther; Wu, Dongmei; Wang, Yangmeng et al. (2018) Sympathetic inputs regulate adaptive thermogenesis in brown adipose tissue through cAMP-Salt inducible kinase axis. Sci Rep 8:11001
Min, Sang-Won; Sohn, Peter Dongmin; Li, Yaqiao et al. (2018) SIRT1 Deacetylates Tau and Reduces Pathogenic Tau Spread in a Mouse Model of Tauopathy. J Neurosci 38:3680-3688
Rauch, Jennifer N; Chen, John J; Sorum, Alexander W et al. (2018) Tau Internalization is Regulated by 6-O Sulfation on Heparan Sulfate Proteoglycans (HSPGs). Sci Rep 8:6382
Masand, Ruchi; Paulo, Esther; Wu, Dongmei et al. (2018) Proteome Imbalance of Mitochondrial Electron Transport Chain in Brown Adipocytes Leads to Metabolic Benefits. Cell Metab 27:616-629.e4
Tekirdag, Kumsal; Cuervo, Ana Maria (2018) Chaperone-mediated autophagy and endosomal microautophagy: Joint by a chaperone. J Biol Chem 293:5414-5424
Theofilas, Panos; Ehrenberg, Alexander J; Nguy, Austin et al. (2018) Probing the correlation of neuronal loss, neurofibrillary tangles, and cell death markers across the Alzheimer's disease Braak stages: a quantitative study in humans. Neurobiol Aging 61:1-12
Kaushik, Susmita; Cuervo, Ana Maria (2018) The coming of age of chaperone-mediated autophagy. Nat Rev Mol Cell Biol 19:365-381
Kampmann, Martin (2018) CRISPRi and CRISPRa Screens in Mammalian Cells for Precision Biology and Medicine. ACS Chem Biol 13:406-416

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