1st Edition

3D Cell-Based Biosensors in Drug Discovery Programs Microtissue Engineering for High Throughput Screening

By William S. Kisaalita Copyright 2010
    404 Pages 109 B/W Illustrations
    by CRC Press

    404 Pages 109 B/W Illustrations
    by CRC Press

    Advances in genomics and combinatorial chemistry during the past two decades inspired innovative technologies and changes in the discovery and pre-clinical development paradigm with the goal of accelerating the process of bringing therapeutic drugs to market. Written by William Kisaalita, one of the foremost experts in this field, 3D Cell-Based Biosensors in Drug Discovery Programs: Microtissue Engineering for High Throughput Screening provides the latest information — from theory to practice — on challenges and opportunities for incorporating 3D cell-based biosensors or assays in drug discovery programs.

    The book supplies a historical perspective and defines the problem 3D cultures can solve. It also discusses how genomics and combinatorial chemistry have changed the way drug are discovered and presents data from the literature to underscore the less-than-desirable pharmaceutical industry performance under the new paradigm. The author uses results from his lab and those of other investigators to show how 3D micro environments create cell culture models that more closely reflect normal in vivo-like cell morphology and function. He makes a case for validated biomarkers for three-dimensionality in vitro and discusses the advantages and disadvantages of promising tools in the search of these biomarkers. The book concludes with case studies of drugs that were abandoned late in the discovery process, which would have been discarded early if tested with 3D cultures.

    Dr. Kisaalita presents evidence in support of embracing 3D cell-based systems for widespread use in drug discovery programs. He goes to the root of the issue, establishing the 3D cell-based biosensor physiological relevance by comparing 2D and 3D culture from genomic to functional levels. He then assembles the bioengineering principles behind successful 3D cell-based biosensor systems. Kisaalita also addresses the challenges and opportunities for incorporating 3D cell-based biosensors or cultures in current discovery and pre-clinical development programs. This book makes the case for widespread adoption of 3D cell-based systems, rendering their 2D counterparts, in the words of Dr. Kisaalita "quaint, if not archaic" in the near future.

    Biosensors and Bioassays
    Conventional Biosensors
    Conventional Biosensor Applications
    Cell-Based Biosensors versus Cell-Based Assays (Bioassays)
    3D Cultures 
    Concluding Remarks

    Target-Driven Drug Discovery
    Drug Discovery and Development
    The Taxol (Paclitaxel) Discovery Case
    The Gleevec (Imatinib Mesylate) Dicovery Case
    Target-Driven Drug Discovery Paradigm
    The New Discovery Paradigm Promise
    Concluding Remarks

    3D versus 2D Cultures
    Comparative Genomics and Proteomics
    Transcriptional Profi ling Studies
    Comparative GO Annotation Analysis
    Proteomics Studies
    Concluding Remarks

    Comparative Structure and Function
    Complex Physiological Relevance
    Cardiomyocyte Contractility
    Liver Cell Bile Canaliculi In Vitro
    Nerve Cell Voltage-Gated Calcium Signaling
    Concluding Remarks

    Emerging Design Principles
    Chemical Microenvironmental Factors
    Cell Adhesion Molecules
    Short-Range Chemistry
    Long-Range Chemistry
    Concluding Remarks

    Spatial and Temporal Microenvironmental Factors
    Nano- and Microstructured Surfaces
    Nano and Scaffold-Combined Structures
    Temporal Factor
    Concluding Remarks

    Material Physical Property and Force Microenvironmental Factors
    Stiffness-Dependent Responses
    Force-Dependent Responses
    Concluding Remarks

    Proteomics as a Promising Tool in the Search for 3D Biomarkers
    Why Search for Three-Dimensionality Biomarkers?
    Cellular Adhesions
    Signaling Pathways
    Overview of Proteomics Techniques
    Study Design and Methods
    Concluding Remarks

    Readout Present and Near Future
    Readout Present and Near Future
    Fluorescence-Based Readouts
    Bioluminescence-Based Readouts
    Label-Free Biosensor Readouts
    Concluding Remarks

    Ready-to-Use Commercial 3D Plates
    Market Opportunities
    Concluding Remarks

    Technology Deployment Challenges and Opportunities
    Challenges to Adopting 3D Cultures in HTS Programs
    Typical HTS Laboratory and Assay Configurations
    Just-in-Time Reagents Provision Model
    Limited Value-Addition from 3D Culture Physiological Relevance: Transepithelium Drug Transport and Induction of Drug Metabolizing Enzyme Cases
    Paucity of Conclusive Support of 3D Culture Superiority

    Cases for 3D Cultures in Drug Discovery
    Three Cases
    The β1-Integrin Monoclonal Antibody Case
    The Matrix Metalloproteinase Inhibitors Case
    Resistance to the Chemotherapeutic Agents Case
    Concluding Remarks

    Ideal Case Study Design
    Rationale for The Case Study
    Why Hepatotoxicity?
    Hepatotoxicity and hESC-Derived Hepatocyte-Like Cells
    Study Design and Methods
    Analysis and Expected Results

    Appendix A:
    Patents for 3D Scaffolds
    Appendix B:
    Current Drug Targets
    Appendix C:
    Popular Cell Lines in Drug Discovery 
    Appendix D:
    Stem Cells in Drug Discovery


    William S. Kisaalita, PhD is professor and former coordinator of graduate engineering programs at the University of Georgia, where he also directs the Cellular Bioengineering Laboratory. The main research focus of his laboratory is cell-surface interactions with applications in cell-based biosensing in drug discovery. He has published more than 80 peer reviewed and trade press papers and made more than 100 poster and podium presentations. He has received numerous instructional awards including membership in the University of Georgia Teaching Academy. He is a member of ACS, AAAS, ASEE, and SBS. Dr. Kisaalita serves on the editorial boards of The Open Biotechnology Journal and The Journal of Community Engagement and Scholarship.