Preface

Chapter 6 **AAV-Mediated Gene Therapy for CRB1-Hereditary**

Celso Henrique Alves and Jan Wijnholds

Chapter 7 **Adeno-Associated Virus (AAV)-Mediated Gene Therapy for Disorders of Inherited and Non-Inherited Origin 139** Indu Rajapaksha, Peter Angus and Chandana Herath

**Retinopathies 119**

**VI** Contents

**Section 3 Cardiac Gene Therapy 167**

Chapter 8 **Gene Therapy for Cardiomyopathies 169** Yves Fromes and Caroline Roques

Gene therapy constitutes a unique class of advanced biological therapy, which works by transferring genetic materials with the aim of regulating, repairing, replacing, adding, or de‐ leting a genetic sequence of interest. This class encompasses nucleic acids transferred using non-viral systems, recombinant viruses that were modified to express a therapeutic gene of interest, genome editing interventions, and genetically engineered human cells that were modified *ex vivo.*

Over the past decades, gene therapy has seen a massive transformation from a proof-of-con‐ cept approach to a clinical reality culminating in the regulatory approval of state-of-the-art products in the European Union and in the United States. These included *in vivo* gene thera‐ py approaches based on recombinant adeno-associated viruses for the treatment of rare in‐ herited genetic disorders such as: lipoprotein lipase deficiency (Glybera®, a replicationdeficient adeno-associated virus serotype 1 (AAV1) expressing S447X variant of human lipoprotein lipase (LPL) gene) and retinal dystrophies caused by mutations in the retinal pigment epithelium-specific 65 KDa (RPE65) gene (Luxturna®, a replication-deficient AAV2 expressing human RPE65 gene). Cancer gene therapy has also dominated the scene, with the approval of cutting-edge gene therapy products. These include the first oncolytic virothera‐ py for melanoma based on attenuated non-integrating Herpes Simplex Virus-1 (HSV-1) modified not only to efficiently replicate within tumors, but also to express the immune stimulatory protein granulocyte macrophage colony-stimulating factor (GM-CSF). Non-sol‐ id tumors, notably B-cell acute lymphoblastic leukaemia (ALL), diffuse large B-cell lympho‐ ma (DLBCL), and primary mediastinal large B-cell lymphoma (PMBCL) have also seen progress in their therapeutic management with the approval of a new generation of *ex vivo* autologous genetically modified T-cell based cancer immunotherapies (Kymriah® and Yes‐ carta®, autologous T-cells genetically engineered with a lentiviral vector (used during Kym‐ riah® manufacturing)/retroviral vector (used during Yescarta® manufacturing) to express anti-CD19 chimeric antigen receptor (CAR)). The clinical and marketing authorization suc‐ cesses of these gene/genetically modified cell-based technologies in cancer and rare genetic diseases have now opened up the pathway for gene therapy application in other new target indications including infections and diabetes.

Gene therapy is continuously shaping and revolutionizing the field of medicine, with more cutting-edge therapies including genome editing-based medicines entering the clinic and be‐ coming a treatment modality in the next 5-10 years.

This book captures some of the scientific progresses notably in gene transfer technologies as well the preclinical and clinical developments of gene therapy interventions (both *ex vivo* and *in vivo*) in the treatment of a broad range of debilitating inherited and non-inherited genetic disorders.

#### **Houria Bachtarzi, DPhil (Oxon), MPharm (Hons), MRPharmS**

ERA Consulting (UK) Ltd A member of the ERA Consulting Group of Companies London Gas Museum, Twelvetrees Crescent London, United Kingdom **Section 1**

**Gene Transfer Technologies for In Vivo and Ex** 

**Vivo Applications**

**Gene Transfer Technologies for In Vivo and Ex Vivo Applications**

This book captures some of the scientific progresses notably in gene transfer technologies as well the preclinical and clinical developments of gene therapy interventions (both *ex vivo* and *in vivo*) in the treatment of a broad range of debilitating inherited and non-inherited

**Houria Bachtarzi, DPhil (Oxon), MPharm (Hons), MRPharmS**

A member of the ERA Consulting Group of Companies

London Gas Museum, Twelvetrees Crescent

ERA Consulting (UK) Ltd

London, United Kingdom

genetic disorders.

VIII Preface

**Chapter 1**

Provisional chapter

**Nucleic Acid-Based Therapy: Development of a**

DOI: 10.5772/intechopen.80741

Gene therapy returns to the center stage of medicine to treat patients with diseases that are unable to be cured with the conventional therapeutic strategies. This development is due to various reasons, including vector development and significant achievement in nextgeneration sequencing. Among the various methodologies of gene therapy, nucleic acidbased therapy has been considered to be promising in various diseases. The development of delivery methods to target cells in vivo, however, remains critical. These include viral vector-based and nonviral vector-based gene delivery methods as well as physical approaches such as hydrodynamic gene delivery (HGD). HGD is a simple and effective in vivo gene transfer method for the functional analysis of therapeutic genes and regulatory elements in small animals. Moreover, this chapter outlines the principle of HGD, gene expression studies in rodents, and recent advances in clinical application of HGD and provides future perspectives in developing a safe and efficient method for nucleic acid-

Keywords: nucleic acid-based therapy, nonviral delivery, hydrodynamic gene delivery,

In 1990, first human gene therapy was conducted, targeting adenosine deaminase deficiency via retrovirus-mediated delivery system [1]. Since then, the number of clinical trials has gradually increased, and approximately 2600 trials have been globally undertaken or approved until November 2017 [2]. Most trials (75%) utilized a viral vector as a delivery tool of gene. Viral vector-based delivery resulted in a high level of gene expression for a long period;

> © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and eproduction in any medium, provided the original work is properly cited.

© 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

site-specificity, computer-controlled injection, human gene therapy

Nucleic Acid-Based Therapy: Development of a

**Nonviral-Based Delivery Approach**

Nonviral-Based Delivery Approach

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

Takeshi Suda and Shuji Terai

Takeshi Suda and Shuji Terai

Abstract

based therapy.

1. Introduction

http://dx.doi.org/10.5772/intechopen.80741

Takeshi Yokoo, Kenya Kamimura, Tsutomu Kanefuji,

Takeshi Yokoo, Kenya Kamimura, Tsutomu Kanefuji,

#### **Chapter 1** Provisional chapter

#### **Nucleic Acid-Based Therapy: Development of a Nonviral-Based Delivery Approach** Nucleic Acid-Based Therapy: Development of a Nonviral-Based Delivery Approach

DOI: 10.5772/intechopen.80741

Takeshi Yokoo, Kenya Kamimura, Tsutomu Kanefuji, Takeshi Suda and Shuji Terai Takeshi Yokoo, Kenya Kamimura, Tsutomu Kanefuji, Takeshi Suda and Shuji Terai

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.80741

#### Abstract

Gene therapy returns to the center stage of medicine to treat patients with diseases that are unable to be cured with the conventional therapeutic strategies. This development is due to various reasons, including vector development and significant achievement in nextgeneration sequencing. Among the various methodologies of gene therapy, nucleic acidbased therapy has been considered to be promising in various diseases. The development of delivery methods to target cells in vivo, however, remains critical. These include viral vector-based and nonviral vector-based gene delivery methods as well as physical approaches such as hydrodynamic gene delivery (HGD). HGD is a simple and effective in vivo gene transfer method for the functional analysis of therapeutic genes and regulatory elements in small animals. Moreover, this chapter outlines the principle of HGD, gene expression studies in rodents, and recent advances in clinical application of HGD and provides future perspectives in developing a safe and efficient method for nucleic acidbased therapy.

Keywords: nucleic acid-based therapy, nonviral delivery, hydrodynamic gene delivery, site-specificity, computer-controlled injection, human gene therapy

#### 1. Introduction

In 1990, first human gene therapy was conducted, targeting adenosine deaminase deficiency via retrovirus-mediated delivery system [1]. Since then, the number of clinical trials has gradually increased, and approximately 2600 trials have been globally undertaken or approved until November 2017 [2]. Most trials (75%) utilized a viral vector as a delivery tool of gene. Viral vector-based delivery resulted in a high level of gene expression for a long period;

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and eproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

however, carcinogenesis and lethal immune reaction were reported [3–5]. Numerous researchers have been attempting to overcome these serious obstacles to enable safe and efficient therapy. For this purpose, the improvement of viral vector has been extensively studied in the last decade, and in addition, nonviral vector-based gene delivery method has developed with great promise. As expected, it resulted in less antigenicity and less chance of integration into the human genome than viral vector; therefore, it can be regarded as a biologically safer method than viral vector-based gene delivery method. However, the period of transgene expression tends to be limited.

This chapter focuses on nonviral vector-based delivery method, which could be used for the nucleic acid-based therapy. In these methods, a transgene is not integrated into the host genome; hence, gene expression is transient. Because temporal transgene expression is applied to promising technologies, such as generation of iPS cells and gene editing by CRISPR/Cas9, nonviral vector-based gene delivery may play a big role in future medicine.

The last section of this chapter outlines the recent progress in the HGD, which enables the highest level of delivery efficiency among nonviral vector-based approaches and the clinical application utilizing the well-established method of catheter insertion into the vessels in the multiple organs.
