Viral vectors for gene delivery and gene therapy within the endocrine system

The transfer of genetic material into endocrine cells and tissues, both in vitro and in vivo, has been identified as critical for the study of endocrine mechanisms and the future treatment of endocrine disorders. Classical methods of gene transfer, such as transfection, are inefficient and limited mainly to delivery into actively proliferating cells in vitro. The development of viral vector gene delivery systems is beginning to circumvent these initial setbacks. Several kinds of viruses, including retrovirus, adenovirus, adeno-associated virus, and herpes simplex virus, have been manipulated for use in gene transfer and gene therapy applications. As different viral vector systems have their own unique advantages and disadvantages, they each have applications for which they are best suited. This review will discuss viral vector systems that have been used for gene transfer into the endocrine system, and recent developments in viral vector technology that may improve their use for endocrine applications - chimeric vectors, viral vector targeting and transcriptional regulation of transgene expression.

Download Full-text

How Far Are Non-Viral Vectors to Come of Age and Reach Clinical Translation in Gene Therapy?

International Journal of Molecular Sciences ◽

10.3390/ijms22147545 ◽

2021 ◽

Vol 22 (14) ◽

pp. 7545

Author(s):

Myriam Sainz-Ramos ◽

Idoia Gallego ◽

Ilia Villate-Beitia ◽

Jon Zarate ◽

Iván Maldonado ◽

...

Keyword(s):

Gene Therapy ◽

Clinical Practice ◽

Gene Delivery ◽

Viral Vectors ◽

Viral Vector ◽

Clinical Success ◽

Genetic Material ◽

Viral Gene ◽

Promising Alternative ◽

Vector Systems

Efficient delivery of genetic material into cells is a critical process to translate gene therapy into clinical practice. In this sense, the increased knowledge acquired during past years in the molecular biology and nanotechnology fields has contributed to the development of different kinds of non-viral vector systems as a promising alternative to virus-based gene delivery counterparts. Consequently, the development of non-viral vectors has gained attention, and nowadays, gene delivery mediated by these systems is considered as the cornerstone of modern gene therapy due to relevant advantages such as low toxicity, poor immunogenicity and high packing capacity. However, despite these relevant advantages, non-viral vectors have been poorly translated into clinical success. This review addresses some critical issues that need to be considered for clinical practice application of non-viral vectors in mainstream medicine, such as efficiency, biocompatibility, long-lasting effect, route of administration, design of experimental condition or commercialization process. In addition, potential strategies for overcoming main hurdles are also addressed. Overall, this review aims to raise awareness among the scientific community and help researchers gain knowledge in the design of safe and efficient non-viral gene delivery systems for clinical applications to progress in the gene therapy field.

Download Full-text

Gene Therapy Approaches

Qubahan Academic Journal ◽

10.48161/qaj.v1n1a35 ◽

2021 ◽

Vol 1 (1) ◽

pp. 52-56

Author(s):

Hogir Saadi

Keyword(s):

Gene Therapy ◽

Viral Vectors ◽

Nuclear Translocation ◽

Viral Vector ◽

Ex Vivo ◽

Genetic Material ◽

Target Tissue ◽

Human Beings ◽

Vector Systems

Gene therapy can be described broadly as the transfer of genetic material to control a disease or at least to enhance a patient's clinical status. The transformation of viruses into genetic shuttles is one of the core principles of gene therapy, which will introduce the gene of interest into the target tissue and cells. To do this, safe strategies have been invented, using many viral and non-viral vector delivery. Two major methods have emerged: modification in vivo and modification ex vivo. For gene therapeutic approaches which are focused on lifelong expression of the therapeutic gene, retrovirus, adenovirus, adeno-associated viruses are acceptable. Non-viral vectors are much less successful than viral vectors, but because of their low immune responses and their broad therapeutic DNA ability, they have advantages. The addition of viral functions such as receptor-mediated uptake and nuclear translocation of DNA may eventually lead to the development of an artificial virus in order to improve the role of non-viral vectors. For human use in genetic conditions, cancers and acquired illnesses, gene transfer techniques have been allowed. The ideal delivery vehicle has not been identified, although the accessible vector systems are capable of transporting genes in vivo into cells. Therefore, only with great caution can the present viral vectors be used in human beings and further progress in the production of vectors is required. Current progresses in our understanding of gene therapy approaches and their delivery technology, as well as the victors used to deliver therapeutic genes, are the primary goals of this review. For that reason, a literature search on PubMed and Google Scholar was carried out using different keywords.

Download Full-text

Gene transfer in the kidney

AJP Renal Physiology ◽

10.1152/ajprenal.1999.276.1.f1 ◽

1999 ◽

Vol 276 (1) ◽

pp. F1-F9 ◽

Cited By ~ 7

Author(s):

Vicki Rubin Kelley ◽

Vikas P. Sukhatme

Keyword(s):

Gene Transfer ◽

Ex Vivo ◽

Future Prospects ◽

Future Directions ◽

Advantages And Disadvantages ◽

Current Vector ◽

Vector Systems ◽

Somatic Gene ◽

Somatic Gene Transfer

Gene transfer approaches offer the promise of revolutionizing medicine. In this review, we focus on the current and future prospects of somatic gene transfer into the kidney. The advantages and disadvantages of current vector systems are described, and the ex vivo and in vitro approaches applicable to the kidney are reviewed. We discuss uses of gene transfer approaches to dissect the pathogenesis of kidney disease and the future directions and applications of gene transfer to combat kidney destruction.

Download Full-text

Gene Therapy: Design and Prospects for Craniofacial Regeneration

Journal of Dental Research ◽

10.1177/0022034509337480 ◽

2009 ◽

Vol 88 (7) ◽

pp. 585-596 ◽

Cited By ~ 29

Author(s):

E.L. Scheller ◽

P.H. Krebsbach

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Gene Delivery ◽

Genetic Material ◽

Building Blocks ◽

Tissue Growth ◽

Viral Gene ◽

Insert Size ◽

Relative Safety ◽

Size Limitation

Gene therapy is defined as the treatment of disease by transfer of genetic material into cells. This review will explore methods available for gene transfer as well as current and potential applications for craniofacial regeneration, with emphasis on future development and design. Though non-viral gene delivery methods are limited by low gene transfer efficiency, they benefit from relative safety, low immunogenicity, ease of manufacture, and lack of DNA insert size limitation. In contrast, viral vectors are nature’s gene delivery machines that can be optimized to allow for tissue-specific targeting, site-specific chromosomal integration, and efficient long-term infection of dividing and non-dividing cells. In contrast to traditional replacement gene therapy, craniofacial regeneration seeks to use genetic vectors as supplemental building blocks for tissue growth and repair. Synergistic combination of viral gene therapy with craniofacial tissue engineering will significantly enhance our ability to repair and replace tissues in vivo.

Download Full-text

Optimizations of In Vitro Mucus and Cell Culture Models to Better Predict In Vivo Gene Transfer in Pathological Lung Respiratory Airways: Cystic Fibrosis as an Example

Pharmaceutics ◽

10.3390/pharmaceutics13010047 ◽

2020 ◽

Vol 13 (1) ◽

pp. 47

Author(s):

Rosy Ghanem ◽

Véronique Laurent ◽

Philippe Roquefort ◽

Tanguy Haute ◽

Sophie Ramel ◽

...

Keyword(s):

Cystic Fibrosis ◽

Gene Therapy ◽

Cell Culture ◽

Gene Transfer ◽

Gene Delivery ◽

Culture Models ◽

Transfer Strategies ◽

Cell Culture Models

The respiratory epithelium can be affected by many diseases that could be treated using aerosol gene therapy. Among these, cystic fibrosis (CF) is a lethal inherited disease characterized by airways complications, which determine the life expectancy and the effectiveness of aerosolized treatments. Beside evaluations performed under in vivo settings, cell culture models mimicking in vivo pathophysiological conditions can provide complementary insights into the potential of gene transfer strategies. Such models must consider multiple parameters, following the rationale that proper gene transfer evaluations depend on whether they are performed under experimental conditions close to pathophysiological settings. In addition, the mucus layer, which covers the epithelial cells, constitutes a physical barrier for gene delivery, especially in diseases such as CF. Artificial mucus models featuring physical and biological properties similar to CF mucus allow determining the ability of gene transfer systems to effectively reach the underlying epithelium. In this review, we describe mucus and cellular models relevant for CF aerosol gene therapy, with a particular emphasis on mucus rheology. We strongly believe that combining multiple pathophysiological features in single complex cell culture models could help bridge the gaps between in vitro and in vivo settings, as well as viral and non-viral gene delivery strategies.

Download Full-text

Nano-Scale Gene Delivery Systems: Current Technology, Obstacles, and Future Directions

Current Medicinal Chemistry ◽

10.2174/0929867325666180108100723 ◽

2018 ◽

Vol 25 (21) ◽

pp. 2448-2464 ◽

Cited By ~ 5

Author(s):

Antonio Garcia-Guerra ◽

Thomas L. Dunwell ◽

Sonia Trigueros

Keyword(s):

Gene Delivery ◽

Physicochemical Properties ◽

Genetic Material ◽

Delivery Systems ◽

Bibliographic Databases ◽

Central Component ◽

Medical Needs ◽

Advantages And Disadvantages ◽

Nano Scale

Within the different applications of nanomedicine currently being developed, nanogene delivery is appearing as an exciting new technique with the possibility to overcome recognised hurdles and several biological and medical needs. The central component of all delivery systems is the requirement for the delivery of genetic material into cells, and for them to eventually reside in the nucleus where their desired function will be exposed. However, genetic material does not passively enter cells; thus, a delivery system is necessary. The emerging field of nano-gene delivery exploits the use of new materials and the properties that arise at the nanometre-scale to produce delivery vectors that can effectively deliver genetic material into a variety of different types of cells. The novel physicochemical properties of the new delivery vectors can be used to address the current challenges existing in nucleic acid delivery in vitro and in vivo. While there is a growing interest in nanostructure-based gene delivery, the field is still in its infancy, and there is yet much to discover about nanostructures and their physicochemical properties in a biological context. We carried out an organised and focused search of bibliographic databases. Our results suggest that despite new breakthroughs in nanostructure synthesis and advanced characterization techniques, we still face many barriers in producing highly efficient and non-toxic delivery systems. In this review, we overview the types of systems currently used for clinical and biomedical research applications along with their advantages and disadvantages, as well as discussing barriers that arise from nano-scale interactions with biological material. In conclusion, we hope that by bringing the far reaching multidisciplinary nature of nano-gene delivery to light, new targeted nanotechnology-bases strategies are developed to overcome the major challenges covered in this review.

Download Full-text

Novel Gene Delivery System of Sendai Virus Transduced Human Peripheral Monocytes.

Blood ◽

10.1182/blood.v108.11.5482.5482 ◽

2006 ◽

Vol 108 (11) ◽

pp. 5482-5482

Author(s):

Tomoko Tanaka ◽

Ryo Kurita ◽

Kashiya Takasugi ◽

Takafumi Nakamura ◽

Makoto Inoue ◽

...

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Gene Delivery ◽

Delivery System ◽

Sendai Virus ◽

Transduction Efficiency ◽

Gene Delivery System ◽

Gene Transduction ◽

Novel Gene

Abstract Extensive monocytes extravasation is seen in infection sites, inflamed tissue and tumor tissues and exhibit a tissue-specific range of functions including phagocytosis, antigen presentation to T cells, and the release of a wide array of cytokines, chemokines, enzymes and nitrogen species. As monocytes have natural tropism to such tissues in each disease condition, these cells can be used as novel gene delivery system to cure the condition. On the other hand, as monocytes are quiescent cells, available gene transfer vectors are limited. Sendai virus (SeV) is a negative-strand RNA virus and has recently been used for the gene therapy vectors, SeV vectors (SeVV), for somatic gene therapy. This vector does not have potential of stable transformation of target cells because they generate no DNA intermediates and therefore are unable to integrate. Also, SeVV has been shown to ensure efficient expression of foreign genes in various types of tissues and can be used for gene transfer into monocytes. We first of all studied the gene transduction efficiency of SeVV into human peripheral blood CD 14 positive monocytes using GFP gene as a marker. About 82% of monocytes became GFP positive, whereas only 23% of lymphocytes and 1.8% of granulocytes became positive. The transduction efficiency was peaked at M.O.I.=1. The expression level was at least preserved for 24 hours in vitro. To do the in vivo preclinical experiments, we also transduced cDNA of GFP and GDNF (glial cell line-derived neurotrophic factor) into the peripheral monocytes of Macaca fascicularis monkey and high level expression of these genes were obtained. In vitro monocyte function after gene transduction are now under investigation. SeVV is considered to be promising gene transfer vector for primate monocytes.

Download Full-text

Silica-Based Gene Delivery Systems: From Design to Therapeutic Applications

Pharmaceutics ◽

10.3390/pharmaceutics12070649 ◽

2020 ◽

Vol 12 (7) ◽

pp. 649 ◽

Cited By ~ 1

Author(s):

Ana Maria Carvalho ◽

Rosemeyre A. Cordeiro ◽

Henrique Faneca

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Targeted Delivery ◽

Viral Vectors ◽

Genetic Material ◽

Successful Outcome ◽

Inorganic Particles ◽

Therapeutic Applications

Advances in gene therapy have been foreshadowing its potential for the treatment of a vast range of diseases involving genetic malfunctioning. However, its therapeutic efficiency and successful outcome are highly dependent on the development of the ideal gene delivery system. On that matter, silica-based vectors have diverted some attention from viral and other types of non-viral vectors due to their increased safety, easily modifiable structure and surface, high stability, and cost-effectiveness. The versatility of silane chemistry and the combination of silica with other materials, such as polymers, lipids, or inorganic particles, has resulted in the development of carriers with great loading capacities, ability to effectively protect and bind genetic material, targeted delivery, and stimuli-responsive release of cargos. Promising results have been obtained both in vitro and in vivo using these nanosystems as multifunctional platforms in different potential therapeutic areas, such as cancer or brain therapies, sometimes combined with imaging functions. Herein, the current advances in silica-based systems designed for gene therapy are reviewed, including their main properties, fabrication methods, surface modifications, and potential therapeutic applications.

Download Full-text

Exosome as a Natural Gene Delivery Vector for Cancer Treatment

Current Cancer Drug Targets ◽

10.2174/1568009620666200924154149 ◽

2020 ◽

Vol 20 (11) ◽

pp. 821-830

Author(s):

Prasad Pofali ◽

Adrita Mondal ◽

Vaishali Londhe

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Nucleic Acids ◽

Cancer Treatment ◽

Intestinal Barrier ◽

Gene Carrier ◽

Gene Delivery Vector ◽

Delivery Vector

Background: Current gene therapy vectors such as viral, non-viral, and bacterial vectors, which are used for cancer treatment, but there are certain safety concerns and stability issues of these conventional vectors. Exosomes are the vesicles of size 40-100 nm secreted from multivesicular bodies into the extracellular environment by most of the cell types in-vivo and in-vitro. As a natural nanocarrier, exosomes are immunologically inert, biocompatible, and can cross biological barriers like the blood-brain barrier, intestinal barrier, and placental barrier. Objective: This review focusses on the role of exosome as a carrier to efficiently deliver a gene for cancer treatment and diagnosis. The methods for loading of nucleic acids onto the exosomes, advantages of exosomes as a smart intercellular shuttle for gene delivery and therapeutic applications as a gene delivery vector for siRNA, miRNA and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and also the limitations of exosomes as a gene carrier are all reviewed in this article. Methods: Mostly, electroporation and chemical transfection are used to prepare gene loaded exosomes. Results: Exosome-mediated delivery is highly promising and advantageous in comparison to the current delivery methods for systemic gene therapy. Targeted exosomes, loaded with therapeutic nucleic acids, can efficiently promote the reduction of tumor proliferation without any adverse effects. Conclusion: In the near future, exosomes can become an efficient gene carrier for delivery and a biomarker for the diagnosis and treatment of cancer.

Download Full-text

Characterization of Recombinant Adeno-Associated Viruses (rAAVs) for Gene Therapy Using Orthogonal Techniques

Pharmaceutics ◽

10.3390/pharmaceutics13040586 ◽

2021 ◽

Vol 13 (4) ◽

pp. 586

Author(s):

Liam Cole ◽

Diogo Fernandes ◽

Maryam T. Hussain ◽

Michael Kaszuba ◽

John Stenson ◽

...

Keyword(s):

Gene Therapy ◽

Light Scattering ◽

Dynamic Light Scattering ◽

Viral Vector ◽

Structural Characteristics ◽

Genetic Material ◽

Label Free ◽

Gene Therapies ◽

Multiple Challenges

Viruses are increasingly used as vectors for delivery of genetic material for gene therapy and vaccine applications. Recombinant adeno-associated viruses (rAAVs) are a class of viral vector that is being investigated intensively in the development of gene therapies. To develop efficient rAAV therapies produced through controlled and economical manufacturing processes, multiple challenges need to be addressed starting from viral capsid design through identification of optimal process and formulation conditions to comprehensive quality control. Addressing these challenges requires fit-for-purpose analytics for extensive characterization of rAAV samples including measurements of capsid or particle titer, percentage of full rAAV particles, particle size, aggregate formation, thermal stability, genome release, and capsid charge, all of which may impact critical quality attributes of the final product. Importantly, there is a need for rapid analytical solutions not relying on the use of dedicated reagents and costly reference standards. In this study, we evaluate the capabilities of dynamic light scattering, multiangle dynamic light scattering, and SEC–MALS for analyses of rAAV5 samples in a broad range of viral concentrations (titers) at different levels of genome loading, sample heterogeneity, and sample conditions. The study shows that DLS and MADLS® can be used to determine the size of full and empty rAAV5 (27 ± 0.3 and 33 ± 0.4 nm, respectively). A linear range for rAAV5 size and titer determination with MADLS was established to be 4.4 × 1011–8.7 × 1013 cp/mL for the nominally full rAAV5 samples and 3.4 × 1011–7 × 1013 cp/mL for the nominally empty rAAV5 samples with 3–8% and 10–37% CV for the full and empty rAAV5 samples, respectively. The structural stability and viral load release were also inferred from a combination of DLS, SEC–MALS, and DSC. The structural characteristics of the rAAV5 start to change from 40 °C onward, with increasing aggregation observed. With this study, we explored and demonstrated the applicability and value of orthogonal and complementary label-free technologies for enhanced serotype-independent characterization of key properties and stability profiles of rAAV5 samples.

Download Full-text