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Biomedical Applications of Proteomics Editedby Jean-CharlesSanchez Garry L. Corthals DenisF. Hochstrasser BiomedicalApplicationofProteomics EditedbyJ.-C.Sanchez,G.Corthals,D.F.Hochstrasser Copyright©2004Wiley-VCHVerlagGmbH&Co.KGaA,Weinheim ISBN:3-527-30807-5 Related Titles from WILEY-VCH ReinerWestermeier,TomNaven Proteomics in Practice A Laboratory Manual of Proteome Analysis 2002, 318 pages ISBN3-527-30354-5 JenniferVanEyk, MichaelJ.Dunn(eds.) Proteomic and Genomic Analysis of Cardiovascular Disease 2003, 396 pages ISBN3-527-30596-3 StefanLorkowski, PaulCullen(eds.) Analysing Gene Expression A Handbook of Methods. Possibilities and Pittfalls 2003, 954 pages ISBN3-527-30488-6 DevKambhampati (ed.) Protein Microarray Technology 2003, 256 pages ISBN3-527-30597-1 Biomedical Applications of Proteomics Edited by Jean-Charles Sanchez Garry L. Corthals Denis F. Hochstrasser Editedby (cid:1) This book was carefully produced. Nevertheless, editors,authorsandpublisherdonotwarrantthe Dr.Jean-CharlesSanchez informationcontainedthereintobefreeofer- Dr.GarryL.Corthals rors.Readersareadvisedtokeepinmindthat Prof.Dr.DenisF.Hochstrasser statements,data,illustrations,proceduraldetails orotheritemsmayinadvertentlybeinaccurate. BiomedicalProteomicsResearchGroup LaboratoireCentraldeChimieClinique HopitauxUniversitairesdeGenève 24,rueMicheli-du-Crest LibraryofCongressCardNo.:appliedfor 1211Genève14 Acataloguerecordforthisbookisavailablefrom Switzerland theBritishLibrary. Bibliographicinformationpublished byDieDeutscheBibliothek DieDeutscheBibliothekliststhispublication intheDeutscheNationalbibliografie;detailed bibliographicdataisavailableintheInternetat <http://dnb.ddb.de>. ©2004WILEY-VCHVerlagGmbH&Co.KGaA, Weinheim Allrightsreserved(includingthoseoftranslation inotherlanguages).Nopartofthisbookmaybe reproducedinanyform–byphotoprinting,micro- film,oranyothermeans–nortransmittedor translatedintomachinelanguagewithoutwritten permissionfromthepublishers.Registerednames, trademarks,etc.usedinthisbook,evenwhennot specificallymarkedassuch,arenottobeconside- redunprotectedbylaw. PrintedintheFederalRepublicofGermany Printedonacid-freepaper Coverdesign SCHULZGrafik-Design, Fußgönheim Composition K+VFotosatzGmbH,Beerfelden Printing betz-druckGmbH,Darmstadt Bookbinding Litges&DopfBuchbindereiGmbH, Heppenheim ISBN 3-527-30807-5 Toour families at home... Anne-Catherine, Christiane and Hélène Jule, Kevin, Lucile,Michael, Rory, Sandrine and Virginie ...and to our BPRG family. BiomedicalApplicationofProteomics EditedbyJ.-C.Sanchez,G.Corthals,D.F.Hochstrasser Copyright©2004Wiley-VCHVerlagGmbH&Co.KGaA,Weinheim ISBN:3-527-30807-5 VII Contents ListofContributors XVII Abbreviations XXV Introduction 1 DenisF. Hochstrasser, Garry L. Corthals, and Jean-CharlesSanchez PartI AspectsinBiomedical Research 5 1 ProteomicsinBiomedicine –ATool,aScience,oranArt? 7 Marc A.Reymond 1.1 Introduction 7 1.2 Diagnosisand Prognosis:an Oxymoron 8 1.3 The Dimensionsof Prognosis 9 1.4 Protein Technologies,Diagnosis,and Prognosis 10 1.5 Individual Protein Patterns in Clinical Practice 10 1.6 New ResearchTools,Old Problems 11 1.7 What isHuman Material? 11 1.8 Using Human Tissuein Biomedical Research– Potential Pitfalls 12 1.9 Informed Consent 12 1.10 Specificity ofProteomicsStudies 14 1.11 Conclusionand Summary 14 1.12 Further Reading 15 PartII Blood Vessels 17 2 Antibody-based VascularTargeting:ProteomicTechniques fortheIdentification andQuantification ofMembraneProteins onEndothelialCells 19 SimoneScheurer, Jascha-NikolaiRybak, ChristophRoesli,Giuliano Elia, and Dario Neri 2.1 Introduction 19 2.2 Vascular Targeting 20 BiomedicalApplicationofProteomics EditedbyJ.-C.Sanchez,G.Corthals,D.F.Hochstrasser Copyright©2004Wiley-VCHVerlagGmbH&Co.KGaA,Weinheim ISBN:3-527-30807-5 VIII Contents 2.2.1 Angiogenesis 20 2.2.2 Angiogenesis-relatedDisorders 21 2.2.3 Markersof Angiogenesis 23 2.2.4 Ligand-basedVascularTargeting 24 2.3 Technologiesforthe Quantitation of MembraneProteinsin Different Cell Types 25 2.3.1 Gel-basedQuantitative Profilingof MembraneProteins 25 2.3.2 Gel-independent Quantitative Profiling ofMembraneProteins 29 2.4 ModelSystems forthe Identification ofVascularTargets 32 2.4.1 In vitro ModelSystems forthe Study ofGeneExpressionin Response to Environmental Changes 32 2.4.2 In vivo ModelSystems forthe Identification of VascularTargets 33 2.5 Conclusions 35 2.6 Acknowledgements 35 2.7 References 36 3 Vasculature,VascularDisease,andAtherosclerosis 39 ElisabettaGianazza and Ivano Eberini 3.1 Introduction 39 3.2 Protein CompositionofHuman Aorta in Atherosclerosis– Ex vivo Studies 41 3.2.1 Cellular Proteins 41 3.2.2 Infiltrating Proteins 42 3.3 Protein CompositionofHuman Aorta in Atherosclerosis– In vitro Studies 44 3.4 Lipoproteinsand Apolipoproteinsas DiseaseFactors 45 3.4.1 Typing of Apolipoprotein E Phenotype in Humans 45 3.4.2 Studiesin Transgenic orKnockout Mice 45 3.5 Pathogenetic Mechanisms 47 3.6 End Pathologies: Myocardialand CerebralInfarction 48 3.7 Surgical Treatments 49 3.8 Pharmacological and Dietary Treatments 49 3.9 Animal Modelsof Atherosclerosisand its Complications 51 3.10 Conclusions 52 3.11 Acknowledgements 52 3.12 References 53 4 Discovery ofNewDiagnostic MarkersofStroke 57 LaureAllard,DenisF. Hochstrasser, and Jean-CharlesSanchez 4.1 Introduction 57 4.2 Stroke Features 57 4.2.1 Brain Anatomy 57 4.2.2 CerebrovascularBloodCirculation 59 4.2.3 Aetiology and Pathology ofStroke 59 4.2.4 Pathophysiology – DifferentTypes of Stroke 59 Contents IX 4.2.5 Epidemiology 60 4.2.6 Treatment 61 4.3 Current Diagnosisof Stroke 61 4.3.1 Physician’s Evaluation 62 4.3.2 Imaging 62 4.3.3 Lumbar Puncture 62 4.3.4 BiochemicalMarkers ofStroke 63 4.4 Proteomic-basedApproach for the Discoveryof Early Diagnostic Stroke Markers 63 4.4.1 One-dimensionalGel Electrophoresis 64 4.4.2 Two-dimensionalGel Electrophoresis 66 4.4.3 SELDI-TOF 69 4.5 Conclusions 70 4.6 Acknowledgements 71 4.7 References 71 PartIII Cancer 73 5 UnravellingBiological PathwaysandtheIdentification ofClinicalMarkers andTargetsinRenalCancer 75 RosamondeE. Banksand PeterJ. Selby 5.1 Renal Cancer – The Clinical Perspective 75 5.1.1 Epidemiology 75 5.1.2 Current Clinical Approachesand Clinical Challenges 76 5.1.3 Immunotherapy for Renal Cancer 78 5.1.4 Existing MarkersorTherapeutic Targets UndergoingClinical Evaluation 79 5.2 ProteomicStudies 80 5.2.1 Tissue-basedStudies 81 5.2.2 Primary and EstablishedCell Lines 87 5.2.3 Biological Fluids 88 5.3 Conclusions 90 5.4 References 91 6 HeatShockProtein27inCancer 97 CeciliaSarto, FulvioMagni, CristinaValsecchi,and Paolo Mocarelli 6.1 Introduction 97 6.2 Genomic Aspects 97 6.3 Structure 98 6.4 Functions 99 6.5 HSP27 Expressionin Cancer 101 6.6 Post-translationalModification and ProteomicTools 102 6.6.1 Phosphorylation 106 6.6.2 S-Thiolation, Oxidation, and Others 106 6.7 Perspectives 107 X Contents 6.8 Acknowledgements 107 6.9 References 108 7 Proteomic ApproachesforBiomarkerDiscovery inColorectal Cancer 111 Richard J. Simpsonand Donna S. Dorow 7.1 Introduction and Background toColorectalCancer 111 7.2 MolecularBasisof ColorectalCancer: Tumorigenesis isa Multistep Process 112 7.3 The CaseforEarly Detection:CRCisTreatableif DetectedEarly 114 7.4 Approachesto Biomarker Discovery 116 7.4.1 UseofELISAs to DetectCRC 119 7.4.2 Two-dimensionalGel Electrophoresis 120 7.4.3 One-dimensionalSodium DodecylSulfate-Polyacrylamide Gel Electrophoresis(SDS-PAGE) Plasma MembraneProteome 123 7.4.4 Multidimensional Protein Identification Technologies 124 7.4.5 ProteomicPattern Diagnostics 125 7.4.6 Protein Microarrays 126 7.4.7 ProteomicApproachesforIdentifyingTumor-specificAutoantigens 127 7.5 Conclusions 127 7.6 References 128 8 ClinicalProteomics: OvarianCancer 133 AyodeleA. Alaiya 8.1 Introduction 133 8.2 General Background 135 8.2.1 OvarianCancer 135 8.2.2 OvarianTumorMarkers 137 8.2.3 ScreeningMethodsandDiagnosticDifficultiesinOvarianTumors 139 8.2.4 Treatment and Prognosisof OvarianTumors 139 8.3 Cancer Proteomics 140 8.3.1 Protein Profiling and Cancer 140 8.3.2 RNA ExpressionAnalysis in CancerCells: Promisesand Pitfalls 141 8.3.3 Potentials and Limitations ofCurrent Protein Profiling Technologies 142 8.3.4 Unravelling Tumor Complexity Prior to ProteomeAnalysis 143 8.3.5 The Future of Clinical Proteomics:Challenges and Opportunities 144 8.4 Short OverviewofOvarian Cancer Proteomics 145 8.4.1 The PromiseofProteomicsin OvarianCancerDiagnostics 145 8.4.2 Analysis ofTissueSamples 146 8.4.3 Analysis ofSerum Samples 147 8.4.4 DiseasePrognosisand Protein ExpressionData 148 8.5 Acknowledgements 150 8.6 References 151 Contents XI 9 ProteinExpressionProfilingAnalysisinHematopoietic StemCells: PhenotypicCharacterization ofMesenchymalStemCells 155 JuanAntonio López,Antonio Bernad, and Juan Pablo Albar 9.1 Introduction 155 9.2 Mesenchymal Stem Cells 155 9.2.1 Stem Cell Definition Criteria 156 9.2.2 New Horizonsin Stem Cell Biology 157 9.2.3 Current and Future Applications of the Stem Cell Technology 157 9.2.4 Stemnessand Stem Cell-associatedGenetic Programs 159 9.3 Proteomics 160 9.3.1 Differential Display via Two-DimensionalGel Electrophoresis 162 9.3.2 Protein Identification 164 9.3.3 Differential Proteomics 164 9.3.4 Protein Profiling 165 9.4 ProteomicAnalysis ofMSCs 165 9.5 Acknowledgements 169 9.6 References 170 10 Lymphoblastoid andLymphoma Cells 173 Raymonde Joubert-Caron, Didier Lutomski,and MichelCaron 10.1 Introduction 173 10.2 Experimental Models 174 10.2.1 Experimental Procedures 174 10.3 Studiesof Protein-pattern Changes Following Treatment with AZC 175 10.3.1 Soluble Protein-pattern Changes Induced by AZC Treatment 177 10.3.2 Membrane-associatedProtein Pattern Changes Induced by AZC Treatment 180 10.4 ProteomicStudy ofGal1-mediated B Cell Apoptosis 183 10.4.1 Identification ofthe MajorGal1-binding MembraneGlycoprotein 184 10.4.2 KineticsofModificationofPhosphorylation ofthe Protein Tyrosine KinaseLyn 184 10.5 Lymphoblastoid and Lymphoma Cells 2-DE Database 186 10.6 References 188 PartIV Pharmaco-toxicology 189 11 Chemoresistance inCancerCells 191 JuliaPoland, DirkSchadendorf, HermannLage,and Pranav Sinha 11.1 Introduction 191 11.2 Two-dimensionalElectrophoresisMaps of GastricCancer, Pancreatic Cancer, and Melanoma 192 11.2.1 GastricCancer 192 11.2.2 PancreaticCancer 194 11.2.3 Melanoma 196

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