The authoritative guide to the revolutionary concept behind the successful Covid-19 vaccines

In Trends in mRNA Vaccine Research, a team of distinguished researchers delivers a practical and up-to-date discussion of the biochemical and biomedical foundations of mRNA vaccines. They also explore the manufacturing conditions required for successful vaccine development and review recent progress in a variety of medical fields, including vaccines against pathogens like SARS-CoV-2, HIV, plasmodium, Mycobacterium tuberculosis, as well as anticancer vaccines.

Volume highlights include:

• A historical overview of mRNA vaccine development
• Immune responses to modified or unmodified mRNA vaccines
• A description of the different mRNA vaccine platforms
• Latest data on current mRNA vaccine developments against infectious diseases and cancer

Perfect for medicinal chemists, immunologists, and epidemiologists, Trends in mRNA Vaccine Research will also benefit researchers and scientists working in the pharmaceutical industry, as well as cancer researchers with an interest in vaccine development.

Preface xiii

Preface from the Volume Editors xv

Part I How mRNA Vaccines Work 1

1 A Historical Overview on mRNA Vaccine Development 3
Rein Verbeke, Miffy H.Y. Cheng, and Pieter R. Cullis

1.1 Introduction 3

1.2 The Path of mRNA as an Unstable and Toxic Product to a New Class of Medicine 5

1.2.1 The Discovery and In Vitro Production of mRNA 5

1.2.2 The Inflammatory Nature of mRNA 7

1.3 How Studying Lipid Bilayer Structures in Cell Membranes Gave Rise to the Eventual Development of Lipid Nanoparticles for RNA Delivery 8

1.3.1 From Biological Cell Membranes to Liposomal Drugs 8

1.3.2 Ionizable Lipid Nanoparticles for Systemic Delivery of Nucleic Acids 10

1.4 The Journey of Developing Clinical mRNA Vaccines 12

1.5 Concluding Remarks 14

References 15

2 Immune Responses to mRNA Vaccine 29
Jean-Yves Exposito, Claire Monge, Danielle C. Arruda, and Bernard Verrier

2.1 Introduction 29

2.2 Innate Sensing of RNA Molecules 30

2.3 Innate Immune Response to mRNA Vaccines 32

2.3.1 Innate Immune Response in Humans 33

2.3.2 Tissue Innate Immune Response in Mice 34

2.4 mRNA Design and Innate Immunity 35

2.4.1 Cap 35

2.4.2 Untranslated Regions 37

2.4.3 Poly(A) 39

2.4.4 Coding Sequence 41

2.5 Optimization and Production of mRNA for an Adequate Innate Immune Response 42

2.5.1 IVT Production 42

2.5.2 Posttranscriptional Modification 44

2.5.3 Purification 45

2.6 mRNA Delivery Systems and Immune Response: The Role of Formulation Composition 45

2.7 Concluding Remarks and Perspectives 51

Acknowledgments 54

References 54

3 Modified or Unmodified mRNA Vaccines? - The Biochemistry of Pseudouridine and mRNA Pseudouridylation 69
Pedro Morais and Yi-Tao Yu

3.1 Pseudouridine ( ): The Fifth Nucleoside 69

3.2 RNA Pseudouridylation Mechanism 70

3.2.1 Naturally Occurring RNA Pseudouridylation 71

3.2.1.1 RNA-independent Pseudouridylation Catalyzed by PUS Enzymes 71

3.2.1.2 RNA-dependent Pseudouridylation Catalyzed by Box H/ACA snoRNP 71

3.2.2 Artificially Introduced RNA Pseudouridylation 73

3.2.2.1 Targeted Pseudouridylation of RNA Using Artificial Guide RNAs 73

3.2.2.2 Incorporation of During In Vitro Transcription of RNA 74

3.3 detectioninRNA 75

3.3.1 Indirect Sequencing Methods 76

3.3.2 Direct Sequencing Methods 76

3.4 Impact of in Pre-mRNA Splicing and Protein Translation 77

3.4.1 Effect of in snRNA and Pre-mRNA on Pre-mRNA Splicing 77

3.4.2 Effect of in rRNA and tRNA on Protein Translation 77

3.4.3 Effect of mRNA Pseudouridylation on Nonsense Suppression 78

3.4.4 Effect of mRNA Pseudouridylation on the Coding Specificity of Sense Codons 80

3.5 and the Immune System 80

3.6 Pseudouridylated Versus Unmodified mRNA Vaccines 82

3.6.1 Successor: N1-methyl- 82

3.6.2 Nucleoside-modified COVID-19 mRNA Vaccines 84

3.6.3 Unmodified mRNA COVID-19 vaccines 85

3.6.4 Cancer mRNA Vaccines 89

3.7 Conclusions 90

Acknowledgments 92

Conflict of Interest 92

References 92

4 Self-Replicating RNA Viruses for Vaccine Development 109
Kenneth Lundstrom

4.1 Introduction 109

4.2 Expression Systems For Self-Replicating RNA Viruses 109

4.3 Vaccines Against Infectious Diseases 113

4.4 Vaccines Against Cancers 130

4.4.1 Reporter Gene Expression 131

4.4.2 Tumor-associated Antigens 131

4.4.3 Cytotoxic and Anti-tumor Genes 139

4.4.4 Immunostimulatory Genes 139

4.4.5 Oncolytic Viruses 140

4.5 Conclusions and Future Aspects 143

References 144

5 Circular RNA Therapeutics and Vaccines 161