Abstract

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.

  Funding Agency

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

  Institution

Name
Wistar Institute
Department
Type
DUNS #
075524595
City
Philadelphia
State
PA
Country
United States
Zip Code
19104

  Publications

Basu, Subhasree; Gnanapradeepan, Keerthana; Barnoud, Thibaut et al. (2018) Mutant p53 controls tumor metabolism and metastasis by regulating PGC-1?. Genes Dev 32:230-243
Seo, Jae Ho; Agarwal, Ekta; Bryant, Kelly G et al. (2018) Syntaphilin Ubiquitination Regulates Mitochondrial Dynamics and Tumor Cell Movements. Cancer Res 78:4215-4228
Lu, Huimin; Bowler, Nicholas; Harshyne, Larry A et al. (2018) Exosomal ?v?6 integrin is required for monocyte M2 polarization in prostate cancer. Matrix Biol 70:20-35
Zhang, Gao; Wu, Lawrence W; Mender, Ilgen et al. (2018) Induction of Telomere Dysfunction Prolongs Disease Control of Therapy-Resistant Melanoma. Clin Cancer Res 24:4771-4784
Wanat, Jennifer J; Logsdon, Glennis A; Driskill, Jordan H et al. (2018) TERRA and the histone methyltransferase Dot1 cooperate to regulate senescence in budding yeast. PLoS One 13:e0195698
Bhattacharjee, Souvik; Coppens, Isabelle; Mbengue, Alassane et al. (2018) Remodeling of the malaria parasite and host human red cell by vesicle amplification that induces artemisinin resistance. Blood 131:1234-1247
Beer, Lynn A; Speicher, David W (2018) Protein Detection in Gels Using Fixation. Curr Protoc Protein Sci 91:10.5.1-10.5.20
Shastrula, Prashanth K; Rice, Cory T; Wang, Zhuo et al. (2018) Structural and functional analysis of an OB-fold in human Ctc1 implicated in telomere maintenance and bone marrow syndromes. Nucleic Acids Res 46:972-984
Wu, Shuai; Fatkhutdinov, Nail; Fukumoto, Takeshi et al. (2018) SWI/SNF catalytic subunits' switch drives resistance to EZH2 inhibitors in ARID1A-mutated cells. Nat Commun 9:4116
Rebecca, Vito W; Nicastri, Michael C; McLaughlin, Noel et al. (2017) A Unified Approach to Targeting the Lysosome's Degradative and Growth Signaling Roles. Cancer Discov 7:1266-1283

Showing the most recent 10 out of 11 publications