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E-raamat: Drug Discovery in Pancreatic Cancer: Models and Techniques

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  • Ilmumisaeg: 11-Mar-2010
  • Kirjastus: Springer-Verlag New York Inc.
  • Keel: eng
  • ISBN-13: 9781441911605
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  • Formaat: PDF+DRM
  • Ilmumisaeg: 11-Mar-2010
  • Kirjastus: Springer-Verlag New York Inc.
  • Keel: eng
  • ISBN-13: 9781441911605
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Pancreatic cancer is the fourth leading cause of cancer death in the United States. Every year, about 33,700 people in the United States will be diagnosed with pancreatic cancer and over 32,000 patients will die from the disease. The median survival of patients with advanced pancreatic cancer is about 6-months. This dismal picture of pancreatic cancer is mainly due to the lack of early diagnosis and effective treatment for patients with advanced disease. To increase the survival rate of pancreatic cancer patients, better tumor markers for diagnosis and new molecular targets for drug development are desperately needed. A lot of effort has been made in searching for pancreatic cancer-causing genes or genes associated with progression of malignant behavior in pancreatic cancer. As a result, alterations in the expression of several cancer-related genes have been identified in pancreatic tumors. The identification and characterization of these cancer-related genes have significantly increased our understanding of pancreatic cancer development, but unfortunately the treatment of pancreatic cancer has not advanced as much in the past 20 years.Over the past decade, tremendous advances have been made in the field of cancer drug discovery, particularly, in the area of molecular and genetic models and technologies. Many of those advanced models and technologies have been applied to the drug discovery processes for pancreatic cancer. In this book, a team of experts will describe the latest development in the application of these models and technologies in pancreatic cancer. The authors include basic researchers as well as clinicians who work in the front-line of the war against pancreatic cancer and have the first-hand experience on these cutting-edge tools and techniques. The book can be divided into two general areas: 1) model systems and 2) genomics and proteomics tools. In recent years there have been a lot of advances in the model systems for pancreatic cancer, including the further characterization of normal and cancerous pancreatic cell lines, the establishment of transgenic mouse models that recapitulate the initiation and progression of human pancreatic cancer, the development of a new xenograft model system for the evaluation of novel agents, and the establishment of a zebrafish pancreatic cancer model. The first four chapters of the book will be devoted to these models. The advances in genomics and proteomics research have made a major impact in cancer drug discovery. A number of these -omics-based tools and techniques have been applied in the pancreatic cancer drug discovery. Chapters 5-9 of the book will discuss techniques for genome-wide examination of gene expression, copy number, methylation, function and regulation. Chapters 10-11 will discuss in situ techniques for studying chromosomal and gene copy number abnormalities as well protein expression changes in cancer samples. Chapters 12-14 will focus on techniques for global examination of protein expression levels in biospecimens obtained from pancreatic cancer patients. Cancer drug discovery has become more and more target-centric.

There have been tremendous advances in cancer drug discovery, and in the area of molecular and genetic models and technologies. In this book, experts describe the latest developments in the application of these models and technologies in pancreatic cancer.
Drug Evaluations in Pancreatic Cancer Culture Systems
1(28)
Bhargava Mullapudi
Yongzeng Ding
Xianzhong Ding
Paul Grippo
Introduction
2(1)
Types of Cells and Cell Lines
2(5)
Normal (Non-Cancerous) Cells
2(1)
Pancreatic Cancer Cell Lines in Culture
3(1)
Rodent Pancreatic Cancer Cell Lines
4(1)
Insulin-Secreting Cell Lines
5(1)
Stem Cells
5(1)
Advantages
6(1)
Limitations
7(1)
Types of Culturing Systems
7(2)
Dish Cultures
7(1)
3D Cultures
8(1)
Co-Cultures
8(1)
High-Throughput Screening
9(1)
Applications for Pharmacotherapeutic Evaluations
9(9)
Signal Transduction Pathways
9(4)
Receptor Tyrosine Kinases
13(2)
VEGF
15(1)
NF-kB (Nuclear Factor-kappaB)
16(2)
Cell Cycle
18(1)
In Vitro Analyses
18(4)
Apoptosis
18(1)
Cell Proliferation
18(1)
DNA Synthesis
19(1)
Cell Viability in Cell Populations
20(1)
Cell Cycle Parameters
21(1)
Invasion/Metastasis
22(1)
Concluding Remarks
22(1)
References
23(6)
Mouse Xenograft Models for Drug Discovery in Pancreatic Cancer
29(22)
Belen Rubio-Viqueira
Manuel Hidalgo
Introduction
29(1)
Classical Drug Development Program at the NCI
30(2)
New Animal Models
32(5)
Orthotopic Tumor Models
32(2)
Genetically Engineered Mouse Models
34(1)
Humanized Mice
35(1)
Freshly Heterotransplanted Human Tumor Xenografts
36(1)
Interpreting Results: Variables and Endpoints
37(3)
Growth Characteristics of the Model
38(1)
Strain of Mice
38(1)
Stage at Which the Treatment Begins
38(1)
The Test Compounds
39(1)
Pharmacokinetics
39(1)
Endpoints
39(1)
Future Approaches in Drug Discovery: Biomarkers and Personalized Therapies
40(4)
Predictive Biomarkers
41(2)
Models and Techniques for Personalized Treatments
43(1)
Concluding Remarks
44(2)
References
46(5)
Fluorescent Metastatic Mouse Models of Pancreatic Cancer for Drug Discovery
51(22)
Michael Bouvet
Robert M. Hoffman
Introduction
52(3)
Green Fluorescent Protein (GFP) Models of Pancreatic Cancer
55(5)
GFP Models of Pancreatic Cancer
55(1)
Tumor Selective Metastatic Organ Targeting
55(2)
Real-Time Simultaneous Whole-Body Imaging of BxPC-3-GFP Tumor and Multiple Metastatic Growth
57(2)
Sequential Intravital Images of Omental and Liver Micrometastasis of BxPC-3-GFP
59(1)
Red Fluorescent Protein (RFP) Models of Pancreatic Cancer
60(5)
Red Fluorescent Protein Models of Pancreatic Cancer
60(1)
Sensitivity of Fluorescence Imaging
61(2)
Use of RFP Models for Drug Discovery and Evaluation
63(2)
Dual-Color Models of Pancreatic Cancer
65(3)
Dual-Color Imaging of Nascent Blood Vessels Vascularizing Pancreatic Cancer in an Orthotopic Model Demonstrates Anti-Angiogenesis Efficacy of Gemcitabine
65(2)
Dual-Color Imaging of Nascent Blood Vessels Vascularizing Liver Metastasis of Pancreatic Cancer also Demonstrates Anti-Angiogenesis Efficacy of Gemcitabine
67(1)
Concluding Remarks
68(1)
References
69(4)
A New Preclinical Paradigm for Pancreas Cancer
73(22)
Sunil R. Hingorani
Introduction
73(3)
Delayed Diagnosis in a Rapidly Lethal Disease
74(1)
Diagnostics and Measures of Treatment Efficacy
74(1)
Lost in Translation
75(1)
Cancer as a Complex Organ
76(1)
Human Disease in a Mammalian Surrogate: Modeling Pancreas Cancer in Mice
77(1)
Lessons Learned from Modeling in Mice
78(3)
Defining a Way Forward
81(1)
Criteria for a Valid Mouse Model of Disease
82(1)
Coordinated Targeting of Stroma and Epithelium: Getting Through the Shield to Hit the Heel
82(4)
Cell Autonomous Targets
82(1)
Non-cell Autonomous Targets
83(3)
Putting Principles into Practice
86(4)
Risk Factors and Chemoprevention
86(1)
Early Detection, Disease Recurrence, and Minimum Residual Disease
86(1)
Target Inhibition
87(1)
Acquired Resistance
87(1)
Imaging
87(1)
Sequencing of Agents and Dosing Schedules
88(2)
Concluding Remarks
90(1)
References
90(5)
Zebrafish as a Biological System for Identifying and Validating Therapeutic Targets and Compounds
95(18)
Nelson S. Yee
Introduction
96(1)
Role of Zebrafish in Pancreatic Cancer
96(1)
Zebrafish Model as a Biological System to Identify Molecular Targets and Validate Drugs
96(1)
Zebrafish Models and Techniques for Drug Discovery in Pancreatic Cancer
97(2)
Wild-Type Zebrafish
99(2)
Exocrine Pancreas of Wild-Type Zebrafish as Models
99(1)
Techniques in Wild-Type Zebrafish for Drug Discovery
100(1)
Germ-Line Mutants
101(3)
Zebrafish with Germ-Line Mutations Affecting Exocrine Pancreas as Models
101(2)
Techniques in Zebrafish with Germ-Line Mutations for Drug Discovery
103(1)
Transgenics
104(2)
Zebrafish Developing Pancreatic Cancer as Models
104(2)
Techniques in Genetically Engineered Zebrafish for Drug Discovery
106(1)
Transplants
106(2)
Zebrafish Xenograft of Pancreatic Cancer as Models
106(2)
Techniques in Zebrafish Xenograft Models for Drug Discovery
108(1)
Concluding Remarks
108(1)
References
109(4)
Gene Expression Arrays in Pancreatic Cancer Drug Discovery Research
113(22)
Charles Gawad
Introduction
113(1)
Gene Expression Profiling Using DNA Microarray
114(2)
Gene Expression Microarray Platforms
114(2)
Technical Considerations
116(1)
Application of Microarray Data in Drug Discovery for Pancreatic Cancer
116(8)
Reclassifying Cancer Types
117(1)
Characterizing Known Cancer Classes
117(2)
Identifying New Classes
119(1)
Expression Signatures of Cancer Subclasses
119(2)
From Lists of Differentially Expressed Genes to Drug Targets
121(2)
Gene Expression Signatures as a Drug Discovery Tool
123(1)
Other Potential Applications of DNA Microarray
124(4)
Biomarkers
124(1)
Drug Resistance
125(1)
Incorporating Gene Expression Arrays with Model Organisms
126(1)
Integrative High-Throughput Analyses
127(1)
Concluding Remarks
128(1)
References
129(6)
Using Array Comparative Genomic Hybridization of Pancreatic Cancer Samples to Map Interesting Regions for Target Gene Identification
135(18)
Chunlao Tang
Robert Lucito
Introduction
135(1)
Alteration of the Genome Is Central to the Causation of All Cancer
136(1)
Development of Array Comparative Genomics Hybridization and Platform Comparison
137(2)
Analysis of Array CGH Data
139(1)
Pancreatic Tumor Samples Analyzed and Findings
140(7)
Further Analysis to Validate Regions and Idenify Cancer Genes
147(2)
Concluding Remarks
149(1)
References
149(4)
The Application of High-Throughput RNAi in Pancreatic Cancer Target Discovery and Drug Development
153(18)
Hongwei Yin
Jeff Kiefer
Michelle Kassner
Nanyun Tang
Spyro Mousses
Introduction
153(1)
RNA Interference (RNAi)
154(2)
RNAi High-Throughput Screening
156(4)
High-Throughput RNAi for Target Identification and Its Application to Pancreatic Cancer
160(4)
High-Throughput RNAi in Drug Discovery and Development and Its Application to Pancreatic Cancer
164(4)
Concluding Remarks
168(1)
References
168(3)
MicroRNA Profilling and Its Application in Drug Discovery in Pancreatic Cancer
171(10)
Aarati R. Ranade
Glen J. Weiss
Introduction
171(1)
miRNA Profiling
172(3)
Microarray Technology
173(1)
Bead-Based Method Using Flow Cytometry
173(1)
Cloning Methods---miRNA Serial Analysis of Gene Expression
174(1)
RNA-Primed Array-Based Klenow Enzyme Assay
175(1)
Study of miRNA in Pancreatic Cancer
175(2)
Application of miRNA Profiling in Drug Discovery in Pancreatic Cancer
177(1)
Concluding Remarks
178(1)
References
178(3)
Methylation Detection and Epigenomics in Pancreatic Cancer
181(24)
Georg Feldmann
Anirban Maitra
Introduction
181(2)
Pancreatic Cancer Is a Genetic Disease
181(2)
Precursor Lesions to Pancreatic Cancer
183(1)
Epigenetic Alterations in Pancreatic Cancer
183(11)
Histone Modifications
184(3)
Global DNA Methylation Studies
187(6)
Epigenetic Regulation of miRNA in Cancer
193(1)
Aberrant DNA Methylation Patterns as Potential Biomarkers of Pancreatic Cancer
193(1)
The Pancreatic Cancer Genome Project
194(1)
Concluding Remarks
195(1)
References
196(9)
Tissue Microarray Applications in Drug Discovery for Pancreatic Cancer
205(18)
Aprill Watanabe
Galen Hostetter
Introduction
205(1)
FFPE Tissues in Cancer Research
206(1)
Tissue Microarrays
207(3)
Types of Tissue Microarrays
208(1)
Construction Equipment
209(1)
Construction Process
210(1)
Representation of Tissue
210(1)
Applications of TMAs
211(2)
Frozen Tissue Arrays
211(1)
Clinical Applications Using TMAs
212(1)
Cell Line Material TMAs
212(1)
In Situ Techniques
213(7)
Immunohistochemistry
214(3)
Fluorescent In Situ Techniques
217(3)
References
220(3)
Proteomic Analysis of Blood and Pancreatic Juice
223(20)
Mark Aspinall-O'Dea
John Neoptolemos
Eithne Costello
Introduction
223(3)
Quantitative LC-MS/MS Approaches
226(5)
Stable Isotope Labelling---In Vitro
226(2)
Stable Isotope Labelling---In Vivo
228(1)
Phosphoproteomics: Detection and Quantification of Phosphoproteins
229(1)
Multiple Reaction Monitoring
230(1)
Overview of Proteomic Analysis of Juice and Blood in Pancreatic Cancer Samples to Date
231(1)
Novel Treatments for Pancreatic Cancer: Analysis of Plasma/Serum Biomarkers
232(4)
The EGFR Pathway in Pancreatic Cancer and Its Therapeutic Intervention: Associated Biomarkers
233(2)
The VEGF Pathway and Its Therapeutic Intervention: Associated Biomarkers
235(1)
Concluding Remarks
236(1)
References
237(6)
Applications of Antibody-Lectin Sandwich Arrays (ALSA) to Pancreatic Cancer Diagnostics and Drug Discovery
243(28)
Yi-Mi Wu
Brian B. Haab
Introduction
243(1)
Protein Glycosylation in Normal Biological Functions
244(5)
Structural Features of Protein Glycosylation
245(1)
N-Glycans
245(2)
O-Glycans
247(2)
Protein Glycosylation in Cancer
249(6)
Roles of Glycans in Cancer Progression
249(2)
Common Glycan Alterations in Cancer
251(4)
Antibody-Lectin Sandwich Arrays (ALSA)
255(4)
Reproducible and Sensitive Detection Using Affinity Reagents
255(2)
Multiplexing Through the Use of Microarrays
257(1)
Convenient Detection of Both Core Protein and Glycan Levels
258(1)
Low-Volume, High-Throughput Sample Processing
258(1)
ALSA in Pancreatic Cancer Research
259(3)
Types of Experiments Using ALSA
259(1)
Example Applications in Pancreatic Cancer Research
260(2)
References
262(9)
The Development of Pharmacodynamic Endpoint Models for Evaluation of Therapeutics in Pancreatic Cancer
271(20)
Amanda F. Baker
Tomislay Dragovich
Introduction
271(1)
Assay Validation
272(1)
In Vitro Models
272(1)
Xenograft Models
273(1)
Hetero-Transplanted Human Xenografts
273(1)
Genetically Engineered Mouse Models
274(1)
Clinical Assessment of PD
274(1)
Imaging Biomarkers
275(1)
PD Models for Gemcitabine
275(4)
Equilibrative Nucleoside Transporter 1
276(1)
Cytidine Deaminase Activity and Deoxycytidylate Deaminase
276(1)
Ribonucleotide Reductase
277(1)
Survivin: An Anti-Apoptotic Gemcitabine Resistance Marker
277(1)
CA 19-9 as a Prognostic Factor for Response to Gemcitabine
278(1)
PD Models for Erlotinib
279(1)
PD Models for Investigational Angiogenesis-Targeting Agents in Pancreatic Cancer
279(2)
Introduction to Angiogenesis in Cancer
279(1)
Angiogenesis as a Therapeutic Target
280(1)
Biomarkers of Angiogenesis in Cancer
280(1)
PD Models Utilized in Assessing Mechanism and Activity of COX-2 Expression and Inhibition in Pancreatic Cancer
281(2)
COX-2 is Highly Expressed in Pancreatic Cancers
281(1)
Selective COX-2 Inhibitors
282(1)
PD Models for COX-2 Inhibition in Pancreatic Cancer
282(1)
Targeting Stroma: Secreted Protein Acid and Rich in Cysteine
283(1)
Other Potential PD Endpoints: Circulating Tumor Cells
283(1)
Concluding Remarks
284(1)
References
284(7)
Index 291