Circulating Tumor Cells

Z. Hugh Fan, PhD, is a professor of the Department of Mechanical and Aerospace Engineering, J. Crayton Pruitt Family Department of Biomedical Engineering, and Department of Chemistry at the University of Florida (UF), USA. Prior to joining UF in 2003, Dr. Fan was a Principal Scientist at ACLARA BioSciences Inc. and was previously a Member of the Technical Staff at Sarnoff Corp. Dr. Fan has been recognized with E.T.S. Walton Award from Science Foundation of Ireland in 2009, Fraunhofer-Bessel Research Award from Alexander von Humboldt Foundation (Germany) in 2010, and UF Research Foundation Professorship in 2014.

List of Contributors xv

Foreword xxi

Preface xxv

PART I INTRODUCTION 1

1 Circulating Tumor Cells and Historic Perspectives 3
Jonathan W. Uhr

1.1 Early Studies on Cancer Dormancy Led to the Development of a Sensitive Assay for CTCs (1970–1998) 3

1.2 Modern Era for Counting CTCs: 1998–2007 6

1.3 Proof of Malignancy of CTCs 7

1.4 New Experiments Involving CTCs 7

1.5 Clinical Cancer Dormancy 8

1.6 Human Epidermal Growth Factor Receptor 2 (HER2) Gene Amplification can be Acquired as Breast Cancer Progresses 10

1.7 uPAR and HER2 Co-amplification 11

1.8 Epithelial–Mesenchymal Transition (EMT) 12

1.9 New Instruments to Capture CTCs 14

1.10 Genotypic Analyses 15

1.11 Conclusions 18

References 20

2 Introduction to Microfluidics 33
Kangfu Chen and Z. Hugh Fan

2.1 Introduction 33

2.2 Scaling Law 36

2.3 Device Fabrication 39

2.4 Functional Components in Microfluidic Devices 43

2.5 Concluding Remarks 46

References 47

PART II ISOLATION METHODS 51

3 Ensemble-decision Aliquot Ranking (eDAR) for CTC Isolation and Analysis 53
Mengxia Zhao, Perry G. Schiro, and Daniel T. Chiu

3.1 Overview of eDAR 53

3.2 Individual Components and Analytical Performance of eDAR 55

3.3 Application and Downstream Analyses of eDAR 69

3.4 Conclusion and Perspective 80

References 81

4 Sinusoidal Microchannels with High Aspect Ratios for CTC Selection and Analysis 85
Joshua M. Jackson, Małgorzata A. Witek, and Steven A. Soper

4.1 Introduction 85

4.2 Parallel Arrays of High-Aspect-Ratio, Sinusoidal Microchannels for CTC Selection 90

4.3 Clinical Applications of Sinusoidal CTC Microchip 114

4.4 Conclusion 118

Acknowledgments 119

References 119

5 Cell Separation using Inertial Microfluidics 127
Nivedita Nivedita and Ian Papautsky

5.1 Introduction 127

5.2 Device Fabrication and System Setup 128

5.3 Inertial Focusing in Microfluidics 129

5.4 Cancer Cell Separation in Straight Microchannels 132

5.5 Cancer Cell Separation in Spiral Microchannels 136

5.6 Conclusions 142

References 142

6 Morphological Characteristics of CTCs and the Potential for Deformability-Based Separation 147
Simon P. Duffy and Hongshen Ma

6.1 Introduction 147

6.2 Limitations of Antibody-based CTC Separation Methods 148

6.3 Morphological and Biophysical Differences Between CTCs and Hematological Cells 149

6.4 Historical and Recent Methods in CTC Separation Based on Biophysical Properties 153

6.5 Microfluidic Ratchet for Deformability-Based Separation of CTCs 155

6.6 Resettable Cell Trap for Deformability-based Separation of CTCs 160

6.7 Summary 165

References 166

7 Microfabricated Filter Membranes for Capture and Characterization of Circulating Tumor Cells (CTCs) 173
Zheng Ao, Richard J. Cote, Ram H. Datar, and Anthony Williams

7.1 Introduction 173

7.2 Size-based Enrichment of Circulating Tumor Cells 174

7.3 Comparison Between Size-based CTC Isolation and Affinity-based Isolation 177

7.4 Characterization of CTCs Captured by Microfilters 178

7.5 Conclusion 180

References 181

8 Miniaturized Nuclear Magnetic Resonance Platform for Rare Cell Detection and Profiling 183
Sangmoo Jeong, Changwook Min, Huilin Shao, Cesar M. Castro,

Ralph Weissleder, and Hakho Lee

8.1 Introduction 183

8.2 μNMR Technology 184

8.3 Clinical Application of μNMR for CTC Detection and Profiling 191

8.4 Conclusion 196

References 196

9 Nanovelcro Cell-Affinity Assay for Detecting and Characterizing Circulating Tumor Cells 201
Millicent Lin, Anna Fong, Sharon Chen, Yang Zhang, Jie-fu Chen, Paulina Do, Morgan Fong, Shang-Fu Chen, Pauline Yang, An-Jou Liang, Qingyu Li, Min Song, Shuang Hou, and Hsian-Rong Tseng

9.1 Introduction 202

9.2 Proof-of-Concept Demonstration of NanoVelcro Cell-Affinity Substrates 207

9.3 First-Generation NanoVelcro Chips for CTC Enumeration 209

9.4 Second-Generation NanoVelcro-LMD Technology for Single CTC Isolation 214

9.5 Third-Generation Thermoresponsive NanoVelcro Chips 219

9.6 Conclusions and Future Perspectives 220

Acknowledgment 221

References 221

10 Acoustophoresis in Tumor Cell Enrichment 227
Per Augustsson, Cecilia Magnusson, Hans Lilja, and Thomas Laurell

10.1 Introduction 227

10.2 Factors Determining Acoustophoresis Cell Separation 230

10.3 Acoustophoresis System for Separating Cells 234

10.4 Acoustophoresis Platform for Clinical Sample Processing 239

10.5 Unperturbed Cell Survival and Phenotype after Microchip Acoustophoresis 244

10.6 Summary 246

References 246

11 Photoacoustic Flow Cytometry for Detection and Capture of Circulating Melanoma Cells 249
John A. Viator, Benjamin S. Goldschmidt, Kiran Bhattacharyya, and Kyle Rood

11.1 Introduction 249

11.2 Current Methods for Detection and Capture of CMCs 254

11.3 Discussion 259

11.4 Future Work 261

References 262

12 Selectin-Mediated Targeting of Circulating Tumor Cells for Isolation and Treatment 267
Jocelyn R. Marshall and Michael R. King

12.1 Introduction 267

12.2 CTC Capture by E-selectin 271

12.3 Applications for E-selectin in Cancer Diagnosis and Treatment 273

12.4 Conclusions 278

References 279

13 Aptamer-Enabled Tumor Cell Isolation 287
Jinling Zhang and Z. Hugh Fan

13.1 Introduction 287

13.2 Aptamers and their Biomedical Applications 288

13.3 Aptamer-based Tumor Cell Isolation 290

13.4 Conclusion and Outlook 297

References 297

14 Depletion of Normal Cells for CTC Enrichment 301
Jeffrey J. Chalmers, Maryam B. Lustberg, Clayton Deighan, Kyoung-Joo Jenny Park, Yongqi Wu, and Peter Amaya

14.1 Introduction 301

14.2 Estimates of Number and Type of Cells in Blood 302

14.3 Summary of Examples of Negative Depletion 303

14.4 Types of Cells Observed After Depletion of Normal Cells 305

14.5 Incomplete Depletion of Normal Cells 305

14.6 Conclusion 310

References 311

PART III POST-ISOLATION ANALYSIS AND CLINICAL TRANSLATION 313

15 Tumor Heterogeneity and Single-cell Analysis of CTCs 315
Evelyn K. Sigal and Stefanie S. Jeffrey

15.1 Introduction 315

15.2 Tumor Heterogeneity 316

15.3 Single-Cell Analysis of CTCs and CTC Heterogeneity 318

15.4 Gene Expression Analysis 319

15.5 Mutational Analysis 321

15.6 Conclusion: Clinical Implications and Future Perspectives 323

References 324

16 Single-Cell Molecular Profiles and Biophysical Assessment of Circulating Tumor Cells 329
Devalingam Mahalingam, Pawel Osmulski, Chiou-Miin Wang, Aaron M. Horning, Anna D. Louie, Chun-Lin Lin, Maria E. Gaczynska, and Chun-Liang Chen

16.1 Introduction 329

16.2 Methods 331

16.3 CTC Applications 336

16.4 Conclusions 342

References 343

17 Directing Circulating Tumor Cell Technologies Into Clinical Practice 351
Benjamin P. Casavant, David Kosoff, and Joshua M. Lang

17.1 Introduction 351

17.2 Defining Biomarkers 352

17.3 The Technology 356

17.4 Translating Technology 357

17.5 Conclusions 360

References 361

PART IV COMMERCIALIZATION 365
18 DEPArray™ Technology for Single CTC Analysis 367

Farideh Z. Bischoff, Gianni Medoro, and Nicolo Manaresi

18.1 Challenges in Molecular Profiling of CTCs 367

18.2 DEPArray™ Technology Solution 368

18.3 DEPArray™ for Single Tumor Cell Analysis 369

18.4 Clinical Significance in Single CTC Profiling 373

18.5 Conclusion 374

References 374

19 CELLSEARCH® Instrument, Features, and Usage 377
Denis A. Smirnov, Brad W. Foulk, Mark C. Connelly, and Robert T. McCormack

19.1 Introduction 377

19.2 Principles of CELLSEARCH® 379

19.3 EpCAM Density and CTC Capture 380

19.4 Clinical Applications of CELLSEARCH® CTCs 383

19.5 Beyond EpCAM Capture 390

19.6 Discussion 391

References 394

PART V GLOSSARY 401

Circulating Tumor Cell Glossary 403
Jose I. Varillas and Z. Hugh Fan

Index 423