Animal Models for the Development of Cancer Immunotherapy

Seng-Lai Tan, PhD, is Chief Scientific Officer at Immunitas Therapeutics. He has more than 20 years of drug discovery and development experience in both small and large molecule therapeutic modalities, including multi-specific antibodies and stapled peptides. He is former Head of Immunology at Marengo (Elstar) Therapeutics and Forma Therapeutics, and has held leadership positions at EMD Serono Research & Development Institute and Roche.

• List of Contributors xiiiIntroduction 1Mithun Khattar and Karuppiah KannanTumor 3Host 5Modality 7References 91 Transplantable Syngeneic Murine Tumor Models 11Rich Woessner, Alexandra Borodovsky, Kris Sachsenmeier, Felix Scheuplein, Robert W. Wilkinson, Chaoyang Ye and Simon Dovedi1.1 Introduction 111.2 Overview of Syngeneic Murine Tumor Models 141.2.1 Immunological Diversity in Syngeneic Models 141.2.2 Technical and Experimental Design Considerations 241.2.2.1 Timeline of Immune Response 241.2.2.2 Lab to Lab Variations 251.2.2.3 Housing and Husbandry Conditions 251.2.2.4 Age and Source of Mice 251.2.2.5 Microbiome Effects 261.2.2.6 Checkpoint Antibodies 261.2.2.7 Site of Implantation 271.3 Modeling the Immune Suppressive Tumor Microenvironment with Syngeneic Models 281.3.1 Immune Suppressive Metabolites 281.3.2 Immune Suppressive Cell Populations 301.3.2.1 Myeloid‐Derived Suppressor Cells 311.3.2.2 Tumor‐Associated Macrophages 321.3.2.3 Regulatory T Cells 331.3.2.4 Regulatory B Cells 341.3.2.5 Concluding Remarks 341.4 Genomic Analysis/Bioinformatics for Syngeneic Tumor Models 351.4.1 Background 351.4.2 Traditional Methods 351.4.2.1 Immunohistochemistry and Immunofluorescence 351.4.2.2 In Silico Imputation 371.4.3 Medium throughput methods 381.4.4 High‐Throughput Methods: scRNAseq 381.4.5 Multi‐omics Methods 391.4.6 Data Analysis 391.4.7 Technical and Sample Considerations 401.4.8 Challenges and Translation into Clinic 401.5 OT‐1 Systems and OVA Expressing Tumor Models for Understanding the Effect of Immuno‐Oncology (I‐O) Agents on T‐Cell Responses 411.6 Modeling Chemotherapy/Radiation Therapy + Immunotherapy Combinations with Syngeneic Models 431.6.1 Preclinical Modeling of Radiotherapy/Immunotherapy Combinations 441.6.2 Preclinical Modeling of Chemotherapy/Immunotherapy Combinations 451.6.3 Summary 461.7 Conclusions 46References 482 Genetically Engineered Mouse Models (GEMMs) in Cancer Immunotherapy R&DZhao Chen 632.1 Current Challenges in Immuno-oncology In vivo Modeling 632.2 Pros and Cons of GEMM in Immuno-oncology Studies 672.3 Organoid-Based GEMM Tumors 682.4 Modeling Different TME Using GEMM 702.5 In vivo Target ID Based on GEMM 712.6 Modeling Immune System Functions with GEMM 71References 723 Mouse Tumor Homografts Recapitulate Human Molecular Pathogenesis 75Rajendra Kumari, Davy Ouyang and Henry Li3.1 Introduction 753.2 Transplanted Syngeneic Cell Line Models 763.3 Spontaneous Autochthonous Mouse Models 783.4 Mouse-Derived Tumor Homografts 813.5 Generation of Tumor Homografts 83Protocol 833.6 Orthotopic Implantation 873.7 Pancreatic Cancer Homografts 883.8 Prostate Cancer Homografts 943.9 Large Scale Mouse Clinical Trial Using Homografts 993.10 Limitations with Regard to Usage of Murine Tumor Homografts 1013.11 Summary 101Acknowledgments 102References 1024 Modeling Resistance to Immune Checkpoint Blockade 107Michael Quigley, Matthew J. Meyer and Kanstantsin Katlinski4.1 Introduction 1074.2 The Use of Preclinical Mouse Models to Understand Mechanisms of Resistance to Checkpoint Blockade 1104.3 Lack of Inflammation 1124.4 Inadequate Antigenicity 1144.5 Suppressive Immune Cell Populations 1174.6 Compensatory T-Cell Checkpoints 1204.7 Suppressive Stromal Factors 1254.8 Tumor-Intrinsic Mechanisms of Resistance to Checkpoint Blockade 1274.9 Identification of New Mechanisms of Resistance and Opportunities for Therapeutic Intervention 1314.10 Conclusion 133References 1345 Orthotopic Mouse Models of Colorectal Cancer and Imaging Techniques 151Ce Yuan, Xianda Zhao, Dechen Wangmo, Travis J. Gates and Subbaya Subramanian5.1 Introduction 1515.2 Orthotopic Model 1525.2.1 Model Establishment 1525.2.2 Surgical Tumor Implantation in the Cecum 1525.2.3 Endoscopy-Guided Intra-colon Wall Tumor Cell Injection 1545.2.4 Immune Features 1545.2.5 Metastasis Features 1555.2.6 Advantages and Limitations 1555.3 Imaging Techniques 1565.3.1 Small Animal Endoscopy 1565.3.2 Magnetic Resonance Imaging 1575.3.3 Positron Emission Tomography 1585.3.4 Imaging Tumor-Specific Markers 1585.3.5 Bioluminescent Imaging (BLI) 1595.3.6 Imaging Immune Cells 1605.4 Summary and Future Directions 160Acknowledgments 162References 1636 Preclinical Osteoimmuno-Oncology Models to Study Effects of Immunotherapies on Bone Metastasis 167Tiina E. Kähkönen, Jussi M. Halleen and Jenni Bernoulli6.1 Introduction to Metastasis and Use of Preclinical Models 1676.2 Bone Metastatic Microenvironment 1696.2.1 Introduction to Bone Biology 1696.2.2 Immune Cells in the Bone Marrow 1706.2.3 Osteoimmunology 1726.2.4 Immunotherapies and Bone Safety 1746.2.5 Vicious Cycle of Bone Metastasis 1756.2.6 Immune Cells in Regulating Metastasis to Bone 1756.2.7 Effects of Estrogens and Androgens on Cancer, Bone, and Immune System 1776.3 Bone Metastasis Models 1786.3.1 Conventional Models to Study Bone Metastasis 1786.3.2 Novel Bone Metastasis Models 1806.3.3 Imaging Techniques in Bone Metastasis Models 1816.3.3.1 Bioluminescence and Fluorescence Imaging and Injectable Dyes 1816.3.3.2 X-ray Imaging 1836.3.3.3 Micro-computed Tomography 1866.3.3.4 Tumor Biomarkers and Bone Turnover Markers 1886.3.3.5 Novel Imaging Techniques 1896.3.3.6 Histology and Immunohistochemistry 1896.3.3.7 Analysis of Immune Cells: Flow Cytometry and Immune Cell Tracking 1916.3.4 What Models to Choose and Readouts to Include in a Bone Metastasis Study? 1936.4 Syngeneic Bone Metastasis Models 1946.4.1 Breast Cancer 1946.4.2 Prostate Cancer 1956.4.3 Multiple Myeloma and Lymphoma 1966.4.4 Osteosarcoma 1976.4.5 Lung and Bladder Cancer 1976.5 Humanized Mouse Models of Bone Metastasis 1986.5.1 Breast Cancer 1996.5.2 Prostate Cancer 2006.6 Conclusions and Future Directions 200References 2017 Humanized Mouse Models for Cancer Immunotherapy: A Focus on Human PBMC and HSPC Reconstitution 211Zhuo Li, Xiao Yang and Xiaomin Song7.1 Immunocompromised Mice 2117.1.1 An Evolving History of Mouse Strains 2127.1.2 Emerging Strain Variants 2137.2 Human Immune System (HIS) Models 2217.2.1 Hu-PBMC Model 2217.2.2 Hu-CD34+ HSPC Model 2267.3 Tumors of Human Origin 2297.3.1 Patient-Derived Xenograft (PDX) 2297.3.2 Conditionally Reprogrammed PDX (CR-PDX) Cells 2307.4 Applications of HIS Mice in I-O Research 2317.4.1 Hu-PBMC Tumor Efficacy Model 2317.4.2 Hu-CD34+ HSPC Tumor Efficacy Model 2327.4.3 Considerations of HIS Tumor Models 2337.4.4 Preclinical Safety Evaluation in HIS Mice 2347.5 Future Perspectives 2357.5.1 Creating Human Tumor Microenvironment (TME) in HIS Mice 2357.5.2 Combining Patient PBMCs and Tumors in HIS Mice 2367.5.3 Human Microbiota-Associated Mice 236References 2378 Bone Marrow–Liver–Thymus (BLT) Humanized Mice as a Tool to Assess Checkpoint Inhibitor Adverse Events 251Kenrick M. Semple, Alan D. Knapton and Kristina E. HowardReferences 2609 Development of Swine Models for Cancer Research: SCID Pigs and Other Emerging Pig Cancer Models 263Adeline N. Boettcher and Christopher K. Tuggle 2639.1 The Emergence of New Swine Biomedical Models 2639.2 Overview of Existing Swine Biomedical Models 2649.2.1 ARTEMIS Deficient SCID Pigs 2649.2.2 RAG1 and RAG2 Knockout SCID Pigs 2659.2.3 IL2RG Knockout SCID Pigs 2669.2.4 Double Mutant SCID Pigs 2669.2.5 Inducible TP53and KRAS Mutant Pigs 2679.3 Overview of Existing and Emerging Cancer Models in Pigs 2679.3.1 B-Cell Lymphoma 2689.3.2 Breast Cancer 2689.3.3 Colorectal Carcinoma 2699.3.4 Glioblastoma 2699.3.5 Hepatocellular Carcinoma 2709.3.6 Melanoma and Histiocytoma 2709.3.7 Osteosarcoma 2719.3.8 Ovarian Cancer 2729.3.9 Pancreatic Cancer 2729.4 Immunotherapy Models to Develop in Pigs 2729.4.1 Humanized SCID Pigs and Xenograft Models 2739.4.2 Transplant Models in Immunocompetent Swine 2749.4.3 Immunotherapeutic Models to Be Developed in Swine Models 2749.4.3.1 Dendritic Cell Vaccines 2749.4.3.2 T- and NK-Cell Therapy Testing 2759.4.3.3 Personalized Xenograft Models in SCID Pigs 2769.4.3.4 Immunometabolism Targeting 2769.5 SCID Pig Housing 2779.5.1 Biocontainment and Biosecurity 2779.5.2 Farrowing, Caesarean-Sections, Piglet Care, and Microbiota 2779.6 Concluding Remarks and Future Directions 278List of Abbreviations 279References 279Index 287