Advanced Technologies in Cardiovascular Bioengineering

Preface
Contents
Part I: Cardiac Development and Morphogenesis
From Simple Cylinder to Four-Chambered Organ: A Brief Overview of Cardiac Morphogenesis
1 Introduction
1.1 Early Gastrulation and Formation of the Cardiac Crescent (E5.0 – E7.5)
1.2 The Linear Heart Tube (E8.0)
1.3 Cardiac Looping (E8.5)
1.4 The Four-Chambered Heart (E9.5)
1.5 The Mature Embryonic Heart (E15.0)
2 Challenges and Opportunities
References
Lineage Tracing Models to Study Cardiomyocyte Generation During Cardiac Development and Injury
1 Introduction
2 Mosaic Analysis with Double Markers (MADM)
2.1 Methodology
2.2 Labeling Rare Populations and Lineage Tracing
2.3 Utility of MADM in Other Organ Systems
2.4 Limitations/Future Directions
3 Rainbow Reporter
3.1 Methodology
3.2 Labeling of Rare Cell Populations and Lineage Tracing
3.3 Utility of Rainbow in Other Organ Systems
3.4 Limitations/Future Directions
References
Mechanisms that Govern Endothelial Lineage Development and Vasculogenesis
1 Cardiovascular Diseases Are Common and Have Considerable Morbidity and Mortality
2 Master Regulators Govern Fate Decisions and Lineage Development
3 Developmental Milestones for Endothelial Development
4 The Common Origin of Endothelial and Hematopoietic Lineages
5 ETV2 Is Necessary and Sufficient for Endothelial Lineage Development
6 ETV2 Expression during Mouse Embryogenesis and the Postnatal Period
7 ETV2 Is Dynamically and Transcriptionally Regulated by Upstream Factors
8 Definition of Transcriptional Targets for ETV2 Mediate Distinct Developmental Events
9 Protein-Protein Interacting Factors for ETV2 Are Important Coregulators
10 ETV2 Is a Master Regulator for Hematoendothelial Lineages Using Conversion Assays
11 ETV2 Overexpression in Tumor Angiogenesis
12 ETV2 Functions to Repress Nonhematoendothelial Fate Decisions
13 Summary
References
Part II: Cellular Approaches to Cardiac Repair and Regeneration
Remuscularization of Ventricular Infarcts Using the Existing Cardiac Cells
1 Introduction
2 Cell Cycle Activation of Existing Cardiomyocytes
2.1 Cardiac Regeneration Achieved with Cardiomyocyte Proliferation
2.2 Approaches to Stimulate Cell Cycle Re-entry of Cardiomyocytes
Cell Cycle Regulators
Signaling Transduction Pathways
Developmental Transcription Factors
2.3 Other Factors
3 Direct Conversion of Cardiac Fibroblasts into Cardiomyocytes
3.1 Reprogramming Factors
3.2 Barriers and Boosters to Direct Cardiac Reprogramming
Small Molecules and Growth Factors
Epigenetic Regulators
Autophagy
Immune Regulators
3.3 Molecular Trajectory of Cardiac Reprogramming
3.4 Direct Cardiac Reprogramming in Human
4 Conclusions and Perspectives
References
Allogeneic Immunity Following Transplantation of Pluripotent Stem Cell-Derived Cardiomyocytes
1 Introduction
1.1 Immunosuppressive Therapies After Heart Transplantation
1.2 Current Immunosuppression Therapies Following Transplantation of PSC-CMs in Clinical Trials
1.3 Allogeneic Transplantation Model in Non-human Primates
1.4 Strategies to Reduce Immunosuppressants Following PSC-CM Transplantation
2 Conclusion
References
Vascular Regeneration with Induced Pluripotent Stem Cell-Derived Endothelial Cells and Reprogrammed Endothelial Cells
1 Introduction
2 Endothelial Cells Generated from Human Pluripotent Stem Cells (hPSC-ECs)
2.1 Derivation of Endothelial Cells from Human Pluripotent Stem Cells
2.2 Remaining Questions and Challenges for hPSC-ECs
3 Direct Reprogramming of Somatic Cells into Endothelial Cells
3.1 Reprogramming into Endothelial Cells by Pluripotent Transcription Factors
3.2 Direct Reprogramming by EC Specific Transcription Factors
3.3 Direct Reprogramming Using Biological Molecules
3.4 Remaining Questions and Challenges for rECs
Clinically Compatibility
Clinical Applicability
4 Future Perspectives
5 Conclusion
References
The Guinea Pig Model in Cardiac Regeneration Research; Current Tissue Engineering Approaches and Future Directions
1 Introduction
2 Tissue Engineered Heart Repair. Cardiac Tissue Engineering Based Regeneration
2.1 How Much Maturation Do We Need?
2.2 Engineered Heart Tissue Transplantation in Small Animal Models
2.3 Upscaling Towards a Clinical Application
2.4 Cell Composition – Do We Need More Than Just Cardiomyocytes?
3 Guinea Pig Model
3.1 Guniea Pig Model in Cardiac Research
3.2 Lessons That Cannot Be Learned from Small Animal Models
3.3 Future Use for the Guinea Pig Model?
Evaluation of the Mode of Action
Improving Transplantation Success
4 Conclusion
References
Part III: Genetic Approaches to Study Cardiac Differentiation and Repair
Analysing Genetic Programs of Cell Differentiation to Study Cardiac Cell Diversification
1 Introduction
2 Designing Stem Cell Biology to Recapitulate Developmental Biology
3 Mesendoderm Patterning and Germ Layer Derivatives
4 Translating Developmental Biology into In Vitro Stem Cell Differentiation Protocols
5 Protocol Development for Cardiovascular Lineage Differentiation in 2D and 3D Systems
5.1 Monolayer and EB-Based Differentiation
5.2 3D and Tissue engineering Based In Vitro Differentiation Protocols
6 Single Cell Technologies Accelerating Discovery into Developmental Biology and Cell Differentiation
7 Cardiac Developmental Single-Cell RNA-Seq Data Analysis
7.1 In Vivo Cardiac Single Cell Analysis
7.2 In Vitro Cardiac Single Cell Analysis
8 Concluding Remarks
References
Recombinant Adeno-Associated Virus for Cardiac Gene Therapy
1 Introduction
2 Gene Therapy Vectors
3 Wild-Type AAV Biology
4 AAV Capsid Biology
5 Cardiac rAAV Tropism in Small Animal Models
6 Cardiac rAAV Tropism in Large Animal Models
7 Therapeutic Evaluation of rAAV Vectors in Large Animal Models of Cardiac Disease
8 Clinical Trials of rAAV-Based Cardiac Gene Therapy
9 Generation of Novel AAV Variants with Increased Cardiotropism
9.1 Liver Detargeting by Ablation of Heparan Binding
9.2 Fusion of a Sodium Ion Channel Ligand to the AAV Capsid
9.3 High Throughput Generation of Novel AAV Capsid Variants
9.4 AAV Capsid Shuffling
10 Rapid Validation of AAV Variants in Heart Tissue
11 Cardiac Promoters to Further Enhance Target Specificity
12 Towards Clinical Translation
13 Conclusions
References
Part IV: Bioengineering Approaches to Cardiovascular Tissue Modeling and Repair
Microfabricated Systems for Cardiovascular Tissue Modeling
1 Introduction
2 Materials for Microfabricated Systems
2.1 Cell Types
Fibroblasts
Endothelial Cells
2.2 Environmental Stimuli
2.3 Extracellular Matrix
2.4 Scaffolds for Microtissues
3 Microfabrication Technology for Cardiovascular Tissue Engineering
3.1 Lithography
3.2 Nonlithography Molding
3.3 3D Printing
3.4 Electrospinning
4 Types of Microfabricated Systems and Platforms
4.1 Biowire I
4.2 Biowire II
4.3 Post/Cantilever Model
4.4 I-Wire Heart-on-a-Chip
4.5 AngioChip
4.6 AngioTube
4.7 2D Flat PDMS Strips
4.8 24-Well Based Platforms
4.9 Microfabricated Cardiac Patches
5 Characterizing Microfabricated Tissues: Incorporating Built-In Readouts into Tissue Platform Design
5.1 Advantages of Microfabricated Platforms for In Vitro Tissue Assessments
5.2 Tissue Contractile Function
5.3 Electrophysiology
5.4 Imaging
5.5 Chemical/Environmental Analyses
6 Applying Microfabricated Systems Towards Challenges in Cardiovascular Research
6.1 Physiological Mechanisms
6.2 Disease Modeling
6.3 Drug Screening and the Discovery of Regenerative Therapeutics
7 Conclusions
References
Bioengineering of Pediatric Cardiovascular Constructs: In Vitro Modeling of Congenital Heart Disease
1 Introduction
2 Procedural Planning and Education
3 Modeling of the Developing Heart
4 Hypoplastic Left Heart Syndrome (HLHS)
5 Pulmonary Artery Stenosis
6 3D Models of Valvular Disease
7 Pump Failure and Drug Delivery
8 Ethical Considerations of 3D Bioprinting and Modeling in Pediatrics
References
Biomaterial Interface in Cardiac Cell and Tissue Engineering
1 Introduction
1.1 Engineering Cardiac Cell and Tissue Microenvironment
1.2 Cell-Biomaterial Interface for Cardiac Tissue Engineering
2 Bulk Hydrogels
2.1 Hydrogel-Based In Vitro 3D Tissue Constructs
2.2 Hydrogel-Based In Vivo Cardiac Therapy
3 Structural Materials
3.1 Porous Materials
3.2 Fibrous Scaffolds
4 Smart Materials
4.1 Smart Materials for In Vitro Dynamic Culture Platforms
4.2 Smart Materials for Drug Delivery Vehicles
5 Conclusion and Future Perspectives
References
Stem Cell-Based 3D Bioprinting for Cardiovascular Tissue Regeneration
1 Introduction
2 Molecular, Cellular and Extracellular Approaches to Promote Cardiovascular Regeneration in Humans
2.1 Cell-Free Approaches
2.2 Cell-Based Approaches
Skeletal Myoblasts (SMs)
Bone Marrow-Derived Cells (BMCs)
Mesenchymal Stem Cells (MSCs)
Cardiac Stem Cells (CSCs)
Embryonic Stem Cells (ESCs)
Induced Pluripotent Stem Cells (iPSCs)
2.3 2D vs 3D Cultures
2.4 Biomaterials for Cardiovascular Tissue Engineering: Polymers, Scaffolds & Hydrogels
Natural Biomaterials
Synthetic Biomaterials
Hybrid Biomaterials
2.5 The Vascularization Problem
2.6 3D Bioprinting of Heart Tissues
In Vitro Testing of 3D Bioprinted Cardiac Tissues
In Vivo Testing of 3D Bioprinted Cardiac Tissues
3 Discussion
4 Conclusions
References
Creating and Validating New Tools to Evaluate the Electrical Integration and Function of hPSC-Derived Cardiac Grafts In Vivo
1 Introduction
2 Evaluation of hPSC-CM Graft EP Function Via Direct Epicardial Surface Recordings
3 Evaluation of hPSC-CM Graft EP Function Via Optical Mapping
3.1 Optical Mapping Using Genetically Encoded Calcium-Sensitive Fluorescent Reporters
3.2 Optical Mapping of GCaMP and a Simultaneously Imaged Water-Soluble Voltage Dye
3.3 Optical Mapping Using Genetically Encoded Voltage-Sensitive Fluorescent Reporters
4 Evaluation of hPSC-CM Graft EP Function in Large-Animal Models
5 Conclusions
References
Part V: Clinical Perspectives
Understanding the Molecular Interface of Cardiovascular Diseases and COVID-19: A Data Science Approach
1 Introduction
2 Materials and Methods
2.1 Unstructured Text Data Sources
2.2 Structured Molecular Data Sources
2.3 CaseOLAP Scoring Platform
3 Results and Discussion
3.1 COVID-19 Symptom-Symptom Relationships
3.2 COVID-19 Symptom-Comorbidity Relationships
3.3 Proteins Implicated in Both COVID-19 and CVDs
All Heart Diseases’ Highest Scoring Proteins
Principal Component Analysis (PCA)
Distinct Nature of CHD and Its Top-Ranking Proteins
3.4 Exploring Relationships Between Associated Proteins in COVID-19 and CVD
Protein Clustering
Exploration of CHD Cluster
Exploration of ARR Cluster
Exploration of OHD Cluster
4 Limitations and Future Work
5 Concluding Remarks
References
Clinical Application of iPSC-Derived Cardiomyocytes in Patients with Advanced Heart Failure
1 Introduction
2 Preparation of Clinical Grade hiPSC-Derived Cardiomyocytes
2.1 Preparation of Clinical Grade hiPSCs
2.2 Large Cell Culture Systems for hiPSC-Derived Cardiomyocyte Expansion
2.3 Purification of hiPSC-Derived Cardiomyocytes
3 Transplant Strategies for hiPSC-Derived Cardiomyocytes
4 Clinical Protocols for Intramyocardial Transplantation of hiPSC-Derived CMs
5 Discussion
5.1 Maturation of Cardiomyocytes
5.2 Immunological Rejection
HLA-Matched iPSCs
Universal Pluripotent Stem Cells
5.3 Preventive Strategies for Tumor Formation
5.4 Electrical Coupling and Arrhythmia
5.5 Implementation in Cardiac Tissues
6 Conclusion
References
Cell Therapy with Human ESC-Derived Cardiac Cells: Clinical Perspectives
1 Why Pluripotent Stem Cells?
1.1 Rationale for the Use of Cardiac Committed Cells
1.2 Interest of PSC as a Source of Cardiac Committed Cells
1.3 The ESCORT Trial
1.4 Other PSC Clinical Trials
1.5 The Issue of Dosing
2 How Do PSC-Derived Cardiac Cells Work?
2.1 The “Remuscularization” Hypothesis
Cell Survival
Arrhythmias
2.2 The Paracrine Hypothesis
Evidence for a Paracrine Mechanism of Action
Use of the PSC Secretome
Advantages
Challenges
2.3 A Common Mechanism-Independent Issue: Safety
3 Perspectives
3.1 Remuscularization
3.2 Paracrine Signalling
References
Index