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 #
5U54NS100717-05
Application #
10011930
Study Section
Special Emphasis Panel (ZNS1)
Project Start
2016-09-30
Project End
2021-08-31
Budget Start
2020-09-01
Budget End
2021-08-31
Support Year
5
Fiscal Year
2020
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

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
Nowakowski, Tomasz J; Rani, Neha; Golkaram, Mahdi et al. (2018) Regulation of cell-type-specific transcriptomes by microRNA networks during human brain development. Nat Neurosci 21:1784-1792
Martinez-Losa, Magdalena; Tracy, Tara E; Ma, Keran et al. (2018) Nav1.1-Overexpressing Interneuron Transplants Restore Brain Rhythms and Cognition in a Mouse Model of Alzheimer's Disease. Neuron 98:75-89.e5
Mavor, David; Barlow, Kyle A; Asarnow, Daniel et al. (2018) Extending chemical perturbations of the ubiquitin fitness landscape in a classroom setting reveals new constraints on sequence tolerance. Biol Open 7:
Caballero, Benjamin; Wang, Yipeng; Diaz, Antonio et al. (2018) Interplay of pathogenic forms of human tau with different autophagic pathways. Aging Cell 17:
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

Showing the most recent 10 out of 15 publications