Target Identification and Validation in Drug Discovery: Methods and Protocols

ISBN-13: 9781627033107 / Angielski / Twarda / 2013 / 371 str.

Jurgen Moll; Riccardo Colombo

Opis

The major reason for the elevated costs of drug development in the pharmaceutical industry is the high attrition rate. In Target Identification and Validation in Drug Discovery: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to identify and validate new target. These include methods and approaches covering biochemical, cell based, in vivo models and translational methods. Chapters also include selected case reports that demonstrate the integration of these technologies to real life experiences and to demonstrate the multiple use of more than one technology to increase knowledge on a specific target. These Written in the highly successful Methods in Molecular Biology series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory.Thorough and intuitive, Target Identification and Validation in Drug Discovery: Methods and Protocols contains a comprehensive list of essential methods and clear protocols to follow.

Spis treści

Part I- Background

 

1. The Path to Oncology Drug Target Validation – an Industry Perspective

 

Marta Cortés-Cros, Tobias Schmelzle, Volker M. Stucke and Francesco Hofmann

 

 

 

Part II- DNA as a tool to Modulate Drug Targets

 

2. Identification of Aptamers as Specific Binders and Modulators of Cell-Surface Receptor Activity

 

Henning Ulrich and Carsten Wrenger

 

 

 

3. The Design and Structure-functional Properties  of DNA-based Immunomodulatory Sequences

 

Nikolai V. Kuznetsov

 

 

 

Part III-RNA Interference : From Design to Data Analysis

 

4. siRNA Design Principles and Off-target Effects

 

Sebastian Petri and Gunter Meister

 

 

 

5. Western Blot Evaluation of siRNA Delivery by pH Responsive Peptides

 

Wanling Liang, A. James Mason and Jenny K.W. Lam

 

 

 

6. High Throughput RNAi Screening for the Identification of Novel Targets

 

Meredith C. Henderson and David O. Azorsa

 

 

 

7. Integration of RNAi and Small Molecule Screens to Identify Targets for Drug Development

 

Konstantinos Drosopoulos and Spiros Linardopoulos

 

 

 

8. CellProfiler and KNIME: Open Source Tools for High Content Screening

 

Martin Stöter, Antje Niederlein, Rico Barsacchi, Felix Meyenhofer, Holger Brandl and Marc Bickle

 

 

 

9. PARP Inhibition as a Prototype for Synthetic Lethal Screens

 

Xuesong Liu

 

 

 

Part IV-Protein Focused Technologies

 

10. Structure-Based Target Druggability Assessment

 

Jean-Yves Trosset and Nicolas Vodovar

 

 

 

11. Validating Pharmacological Disruption of Protein-protein Interactions by Acceptor Photobleaching FRET Imaging

 

Janos Roszik, Gábor Tóth, János Szöllősi and György Vereb

 

 

 

12. Systematic Analysis of Complex Signal Transduction Pathways using Protein Fragment Complementation Assays

 

Thomas I. Koblizek, Ann Siehoff and Anthony Pitt

 

 

 

13. Reverse-Phase Protein Microarrays and Their Utility in Drug Development

 

Maria Isabella Sereni , Mariaelena Pierobon,Roberto Angioli Emanuel F. Petricoin III, and Mitchell J. Frederick

 

 

 

Part V. Innovative Cell Culture Techniques to Mimic Tissue Microenvironments

 

14. A Cell Culture System that Mimics Chronic Lymphocytic Leukemia Cells Microenvironment for Drug Screening and Characterization

 

Alessandro Natoni

 

 

 

15. 2-Dimensional vs. 3-Dimensional in vitro Tumor Migration and Invasion Assays

 

Miriam Zimmermann, Carol Box and Suzanne A. Eccles

 

 

 

16. Tumor Spheroid-based Migration Assays for Evaluation of Therapeutic Agents

 

Maria Vinci, Carol Box, Miriam Zimmermann and Suzanne A. Eccles

 

 

 

17. The NeuroSphere Assay (NSA) Applied to Neural Stem Cells (NSCs) and Cancer Stem Cells (CSCs)

 

Rossella Galli

 

 

Part VI- Genetically Engineered Animal Models to study Gene Functions

 

18. Genetically Engineered Animal Models for in vivo Target Identification and Validation in Oncology

Gemma Texidó

 

 

 

19. Target Validation in Mice by Constitutive and Conditional RNAi

 

Aljoscha Kleinhammer, Wolfgang Wurst, Ralf Kühn

 

 

 

20. In vivo Target Validation by Inducible RNAi in Human Xenograft Mouse Models

 

Marco Mazzoletti and Gemma Texido

 

 

 

Part VII- Translational Methods to validate Biomarkers

 

21. Brightfield In Situ Hybridization Methods to Discover Gene Amplifications and Rearrangements in Clinical Samples

 

Hiroaki Nitta and Thomas M. Grogan

 

 

 

22. Combined microRNA In Situ Hybridization and Immunohistochemical Detection of Protein Markers

 

Boye Schnack Nielsen and Kim Holmstrøm

 

 

 

Thomas I. Koblizek, Ann Siehoff and Anthony Pitt