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 #
5U54NS100717-02
Application #
9360024
Study Section
Special Emphasis Panel (ZNS1)
Project Start
Project End
2018-08-31
Budget Start
2017-09-01
Budget End
2018-08-31
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost

  Institution

Name
J. David Gladstone Institutes
Department
Type
DUNS #
099992430
City
San Francisco
State
CA
Country
United States
Zip Code
94158

  Publications

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
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:

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