TheProteomicsandMetabolomicsFacility,supportedbytheCancerCenterSupportGrant(CCSG)awarded toTheWistarInstitute,providesacomprehensivesetofproteomicsandtargetedquantitativemetabolite assaystotheWistarCancerCentermembershipasaprimarygoal.ResourcesofthisFacilityarealso availabletoinvestigatorsinotherCancerCentersandotheracademicinvestigatorsasasecondarygoal.Dr. Tang?sroleastheFacilityManagingDirectoristosupportcancerresearchandotherbiomedicalresearchby providingexpertconsultationandstate-of-the-arttechnologiesthatoperateatmaximumperformanceat affordablecoststousers.TheManagingDirectorwillassistinexperimentaldesign,manageFacilitystaff, performMSdataanalysesasneeded,andassistinthebiologicalinterpretationofresults.TheManaging Directorwillalsodevotesubstantialefforttooptimizingandimplementingnewmethods,updateanalyticaland dataanalysesmethods,andupdateinstrumentationtoensureeachprojectisperformedusingstate-of-the-art methodologies.Thisiscriticalbecauseinstrumentation,software,andanalyticalstrategiescontinuetoevolve rapidly,andmostcurrentandanticipatedfutureprojectsinvolveverychallengingproteomicsand metabolomicsproblems.Proteomicsprojectswillinclude:1)LC-MS/MSanalysisofisolatedproteincomplexes withandwithoutchemicalcrosslinking,2)in-depth,global,quantitativecomparisonsofexosomes,secretomes, andcelllysates,and3)targetedandglobalquantitativecomparisonsofposttranslationalmodifications. Quantitativedatawillbeobtainedeitherusinglabel-freequantitationofintegratedMSioncurrents,or quantitationusingstableisotopessuchasSILACorisobarictag(TMToriTRAQ)labeling.Theproteome analyseswilluseoptimizedmethodsandstate-of-the-artinstruments,suchastheThermoScientificQ ExactiveHFmassspectrometer,thatcandetectandrobustlyquantifymostoftheproteinspresentincomplex samples,includingwholecelllysates.Metabolomicsprojectswillincludequantifyingthesteadystatelevelsof keymetabolitesusingtargetedmultiplexMRM-MSontheSciexQTRAP5500hybridtriplequadrupolelinear iontrapmassspectrometer.Plansforfuturedevelopmentinclude:1)expandingdepthofproteomeanalyses toefficientlyanalyzegreaterthan90%ofgeneproductsexpressedbycellsandtissues,2)implementing routineuseofchemicalcross-linkingtoidentifyprotein-proteininteractionsandobtainstructuralinformation,3) implementationofmetabolitefluxanalysesusing13Cstableisotopetracer,and4)implementationoflipidomics analyses.AllofthesenewmethodswillusetheThermoScientificQExactivemassspectrometersorfuture nextgenerationinstruments.Theseproteomicsandmetabolomicsanalyseswillcontributetocriticaldata requiredtoidentifyproteinandmetabolitetargets,aswellasgeneratehypothesesthatarevitalforthesuccess ofthecancer-relatedprojectsdescribedinthisapplication.

Public Health Relevance

Thisapplicationwillsupportcancerandotherbiomedical-relatedresearcheffortsofWistarInstitute investigatorsbyprovidinghighlevel,cuttingedgeproteomicsandmetabolomicsexpertiseandanalytical capacitiesthroughtheProteomicsandMetabolomicsFacility.Focusofthesupportedprojectsinvolve experimentsoncancercelllines,primarytumors,animalmodels,andpatientbiologicalfluids.Themajoraims oftheseprojectsaretoidentifycomponentsofproteincomplexestohelpunderstandtheirfunctions,elucidate posttranslationalmodifications,identifysignificantlychangingproteinormetabolitetargets,andtoquantify targetsindifferenttypeofsamples.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Project #
1R50CA221838-01
Application #
9422013
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Knowlton, John R
Project Start
2017-09-20
Project End
2022-08-31
Budget Start
2017-09-20
Budget End
2018-08-31
Support Year
1
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Wistar Institute
Department
Type
DUNS #
075524595
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
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