Niosome-Based Approach for In Situ Gene Delivery to Retina and Brain Cortex as Immune-Privileged Tissues

Non-viral vectors have emerged as a promising alternative to viral gene delivery systems due to their safer profile. Among non-viral vectors, recently, niosomes have shown favorable properties for gene delivery, including low toxicity, high stability, and easy production. The three main components of niosome formulations include a cationic lipid that is responsible for the electrostatic interactions with the negatively charged genetic material, a non-ionic surfactant that enhances the long-term stability of the niosome, and a helper component that can be added to improve its physicochemical properties and biological performance. This review is aimed at providing recent information about niosome-based non-viral vectors for gene delivery purposes. Specially, we will discuss the composition, preparation methods, physicochemical properties, and biological evaluation of niosomes and corresponding nioplexes that result from the addition of the genetic material onto their cationic surface. Next, we will focus on the in situ application of such niosomes to deliver the genetic material into immune-privileged tissues such as the brain cortex and the retina. Finally, as future perspectives, non-invasive administration routes and different targeting strategies will be discussed.

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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.

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Current Designs of Polymeric Platforms towards the Delivery of Nucleic Acids inside the Cells with Focus on Polyethylenimine

Current Gene Therapy ◽

10.2174/1566523221666210705130238 ◽

2021 ◽

Vol 21 ◽

Author(s):

Fernando A. de Oliveira ◽

Lindomar J. C. Albuquerque ◽

Gwendoline Delecourt ◽

Véronique Bennevault ◽

Philippe Guégan ◽

...

Keyword(s):

Gene Delivery ◽

Nucleic Acids ◽

Viral Vectors ◽

Transfection Efficiency ◽

Imaging Techniques ◽

Structural Parameters ◽

Genetic Material ◽

Basic Research ◽

Viral Gene ◽

Gene Delivery Vectors

Background: Gene delivery is a promising technology for treating diseases linked to abnormal gene expression. Since nucleic acids are the therapeutic entities in such approach, a transfecting vector is required because the macromolecules are not able to efficiently enter the cells by themselves. Viral vectors have been evidenced to be highly effective in this context; however, they suffer from fundamental drawbacks, such as the ability to stimulate immune responses. The development of synthetic vectors has accordingly emerged as an alternative. Objectives: Gene delivery by using non-viral vectors is a multi-step process that poses many challenges, either regarding the extracellular or intracellular media. We explore the delivery pathway and afterwards, we review the main classes of non-viral gene delivery vectors. We further focus on the progresses concerning polyethylenimine-based polymer-nucleic acid polyplexes, which have emerged as one of the most efficient systems for delivering genetic material inside the cells. Discussion: The complexity of the whole transfection pathway, along with a lack of fundamental understanding, particularly regarding the intracellular trafficking of nucleic acids complexed to non-viral vectors, probably justifies the current (beginning of 2021) limited number of formulations that have progressed to clinical trials. Truly, successful medical developments still require a lot of basic research. Conclusion: Advances in macromolecular chemistry and high-resolution imaging techniques will be useful to understand fundamental aspects towards further optimizations and future applications. More investigations concerning the dynamics, thermodynamics and structural parameters of polyplexes would be valuable since they can be connected to the different levels of transfection efficiency hitherto evidenced.

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Polyplex: A Promising Gene Delivery System

International Journal of Pharmaceutical Sciences and Nanotechnology ◽

10.37285//ijpsn.2019.12.6.1 ◽

2019 ◽

Vol 12 (6) ◽

pp. 4681-4686

Author(s):

Rohan Aggarwal ◽

Monika Targhotra ◽

Bhumika Kumar ◽

P.K Sahoo ◽

Meenakshi K Chauhan

Keyword(s):

Gene Transfer ◽

Gene Delivery ◽

Delivery System ◽

Viral Vectors ◽

Gene Delivery System ◽

Cationic Polymers ◽

Therapeutic Application ◽

Promising Alternative ◽

Structure And Stability ◽

The Past

In the past few years gene delivery system has gained a huge attention owing to its proved efficacy in several diseases especially in those caused by genetic and/oroncological malfunctioning. The effective gene delivery mainly depends on the carrier molecules that can ensure the safe and specific delivery of the nucleic acidmolecules. Viral vectors have been used for a longer period as the gene transfer vehicle. However, these viral vectors have potential immunological disadvantages that made them less preferred. Recently, non-viral vectors such as polyplexes have emerged as a promising alternative for viral vectors. Polyplexes are formed by conjugating a polymer with DNA and in maximum cases the cationic polymers are preferred over others. The structure and stability of the polyplexes depends on various factors. The ability of the polymer to condense the DNA mainly dictates the efficiency of the polyplex mediated transfection. In this review we are going to provide a framework for the synthesis and design of the polyplexes along with the structure and stability of the complexes pertaining to mechanism of action, characterization and therapeutic application, including polyethyleneimine mediated cytotoxicity as well as newer strategies for the generation of better polyplexes.

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Aliphatic Quaternary Ammonium Functionalized Nanogels for Gene Delivery

Pharmaceutics ◽

10.3390/pharmaceutics13111964 ◽

2021 ◽

Vol 13 (11) ◽

pp. 1964

Author(s):

Huaiying Zhang ◽

Damla Keskin ◽

Willy H. de Haan-Visser ◽

Guangyue Zu ◽

Patrick van Rijn ◽

...

Keyword(s):

Gene Delivery ◽

Tertiary Amine ◽

Quaternary Ammonium ◽

Gene Transfection ◽

Genetic Material ◽

Cationic Lipid ◽

Endosomal Escape ◽

Hereditary Diseases ◽

Viral Gene

Gene therapy is a promising treatment for hereditary diseases, as well as acquired genetic diseases, including cancer. Facing the complicated physiological and pathological environment in vivo, developing efficient non-viral gene vectors is needed for their clinical application. Here, poly(N-isopropylacrylamide) (p(NIPAM)) nanogels are presented with either protonatable tertiary amine groups or permanently charged quaternized ammonium groups to achieve DNA complexation ability. In addition, a quaternary ammonium-functionalized nanogel was further provided with an aliphatic moiety using 1-bromododecane to add a membrane-interacting structure to ultimately facilitate intracellular release of the genetic material. The ability of the tertiary amine-, quaternized ammonium-, and aliphatic quaternized ammonium-functionalized p(NIPAM) nanogels (i.e., NGs, NGs-MI, and NGs-BDD, respectively) to mediate gene transfection was evaluated by fluorescence microscopy and flow cytometry. It is observed that NGs-BDD/pDNA complexes exhibit efficient gene loading, gene protection ability, and intracellular uptake similar to that of NGs-MI/pDNA complexes. However, only the NGs-BDD/pDNA complexes show a notable gene transfer efficiency, which can be ascribed to their ability to mediate DNA escape from endosomes. We conclude that NGs-BDD displays a cationic lipid-like behavior that facilitates endosomal escape by perturbing the endosomal/lysosomal membrane. These findings demonstrate that the presence of aliphatic chains within the nanogel is instrumental in accomplishing gene delivery, which provides a rationale for the further development of nanogel-based gene delivery systems.

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A ligand-based system for receptor-specific delivery of proteins

Scientific Reports ◽

10.1038/s41598-019-55797-1 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Mariano Maffei ◽

Chiara Morelli ◽

Ellie Graham ◽

Stefano Patriarca ◽

Laura Donzelli ◽

...

Keyword(s):

Gene Delivery ◽

Protein Delivery ◽

Genetic Diseases ◽

Cell Surface Receptor ◽

Viral Gene ◽

Promising Alternative ◽

Large Proteins ◽

Reporter Mice ◽

Surface Receptor

AbstractGene delivery using vector or viral-based methods is often limited by technical and safety barriers. A promising alternative that circumvents these shortcomings is the direct delivery of proteins into cells. Here we introduce a non-viral, ligand-mediated protein delivery system capable of selectively targeting primary skin cells in-vivo. Using orthologous self-labelling tags and chemical cross-linkers, we conjugate large proteins to ligands that bind their natural receptors on the surface of keratinocytes. Targeted CRE-mediated recombination was achieved by delivery of ligand cross-linked CRE protein to the skin of transgenic reporter mice, but was absent in mice lacking the ligand’s cell surface receptor. We further show that ligands mediate the intracellular delivery of Cas9 allowing for CRISPR-mediated gene editing in the skin more efficiently than adeno-associated viral gene delivery. Thus, a ligand-based system enables the effective and receptor-specific delivery of large proteins and may be applied to the treatment of skin-related genetic diseases.

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Long-Circulating Polymeric Nanovectors for Tumor-Selective Gene Delivery

Technology in Cancer Research & Treatment ◽

10.1177/153303460500400605 ◽

2005 ◽

Vol 4 (6) ◽

pp. 615-625 ◽

Cited By ~ 71

Author(s):

Sushma Kommareddy ◽

Sandip B. Tiwari ◽

Mansoor M. Amiji

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Viral Vectors ◽

High Efficiency ◽

Transfection Efficiency ◽

The United States ◽

Medical Intervention ◽

Hydrophilic Polymers ◽

Circulation Time ◽

Viral Gene

Significant advances in the understanding of the genetic abnormalities that lead to the development, progression, and metastasis of neoplastic diseases has raised the promise of gene therapy as an approach to medical intervention. Most of the clinical protocols that have been approved in the United States for gene therapy have used the viral vectors because of the high efficiency of gene transfer. Conventional means of gene delivery using viral vectors, however, has undesirable side effects such as insertion of mutational viral gene into the host genome and development of replication competent viruses. Among non-viral gene delivery methods, polymeric nanoparticles are increasingly becoming popular as vectors of choice. The major limitation of these nanoparticles is poor transfection efficiency at the target site after systemic administration due to uptake by the cells of reticuloendothelial system (RES). In order to reduce the uptake by the cells of the RES and improve blood circulation time, these nanoparticles are coated with hydrophilic polymers such as poly(ethylene glycol) (PEG). This article reviews the use of such hydrophilic polymers employed for improving the circulation time of the nanocarriers. The mechanism of polymer coating and factors affecting the circulation time of these nanocarriers will be discussed. In addition to the long circulating property, modifications to improve the target specificity of the particles and the limitations of steric protection will be analyzed.

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Recent Advances in the Development of Bio-Reducible Polymers for Efficient Cancer Gene Delivery Systems

10.46619/cmj.2019.2-1007 ◽

2019 ◽

Vol 2 (1) ◽

pp. 6-13 ◽

Cited By ~ 1

Author(s):

Kiel Sung Yong ◽

◽

Wan Kim Sung ◽

◽

...

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Ribonucleic Acid ◽

Viral Vectors ◽

Delivery Systems ◽

Host Cells ◽

Viral Gene ◽

Cancer Gene ◽

Inherited Disorders ◽

Messenger Ribonucleic Acid

Gene therapy is the unique method for the use of genetic materials such as Messenger ribonucleic acid (mRNA), plasmid deoxyribonucleic acid (pDNA), and small interfering ribonucleic acid (siRNA) into specific host-cells for the treatment of inherited disorders in any diseases. The successful way to utilize the gene therapy is to develop the efficient cancer gene delivery systems. In this paper, the successful and efficient gene delivery systems are briefly reviewed on the basis of bio-reducible polymeric systems for cancer therapy. The viral gene delivery systems such as RNA-based viral and DNA-based viral vectors are also discussed. The development of bio-reducible polymer for gene delivery system has briefly discussed for the efficient cancer gene delivery of viral vectors and non-viral vectors.

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Genetic modification of mesenchymal stem cells to enhance their anti-tumor efficacy

Asia-Pacific Journal of Oncology ◽

10.32948/ajo.2021.03.28 ◽

2021 ◽

pp. 1-6

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Gene Delivery ◽

Genetic Modification ◽

Chemokine Receptors ◽

Viral Vectors ◽

Viral Gene ◽

Delivery Methods ◽

Differentiation Potential ◽

Viral Gene Delivery

Non-hematopoietic mesenchymal stem cells (MSCs) are widely used in regenerative medicine and tissue engineering as they possess multilineage differentiation potential and self-renewal properties. MSCs can be easily isolated from several tissues and expanded following standard cell culture procedures. MSCs have the capability of mobilization to the tumor site; so, they can automatically relocate to the tumor sites through their chemokine receptors following intravenous transplantation. In this respect, they can be used for MSC-based gene therapy. In this therapeutic technique, beneficial genes are inserted by viral and non-viral methods into MSCs that lead to transgene expression in them. Genetic modifications of MSCs have been widely studied and thoroughly investigated to further enhance their therapeutic efficacy. The current strategies of MSC-based therapies emphasize the incorporation of beneficial genes, which will enhance the therapeutic ability of MSCs and have better homing efficiency. Non-viral methods produce less toxicity and immunogenicity compared to viral gene delivery methods and thus represent a promising and efficient tool for the genetic engineering of MSCs. Several non-viral gene delivery strategies have been developed in recent decades, and some of them have been used for MSCs modification. This mini review provides an overview of current gene delivery approaches used for the genetic modification of MSCs with beneficial genes including viral and non-viral vectors.

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Polymeric Nanoparticles in Gene Therapy: New Avenues of Design and Optimization for Delivery Applications

Polymers ◽

10.3390/polym11040745 ◽

2019 ◽

Vol 11 (4) ◽

pp. 745 ◽

Cited By ~ 41

Author(s):

Raj Rai ◽

Saniya Alwani ◽

Ildiko Badea

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Targeted Delivery ◽

Polymeric Nanoparticles ◽

Genetic Material ◽

Delivery Systems ◽

Viral Gene ◽

Cationic Polymers ◽

Natural Polymers ◽

Viral Gene Delivery

The field of polymeric nanoparticles is quickly expanding and playing a pivotal role in a wide spectrum of areas ranging from electronics, photonics, conducting materials, and sensors to medicine, pollution control, and environmental technology. Among the applications of polymers in medicine, gene therapy has emerged as one of the most advanced, with the capability to tackle disorders from the modern era. However, there are several barriers associated with the delivery of genes in the living system that need to be mitigated by polymer engineering. One of the most crucial challenges is the effectiveness of the delivery vehicle or vector. In last few decades, non-viral delivery systems have gained attention because of their low toxicity, potential for targeted delivery, long-term stability, lack of immunogenicity, and relatively low production cost. In 1987, Felgner et al. used the cationic lipid based non-viral gene delivery system for the very first time. This breakthrough opened the opportunity for other non-viral vectors, such as polymers. Cationic polymers have emerged as promising candidates for non-viral gene delivery systems because of their facile synthesis and flexible properties. These polymers can be conjugated with genetic material via electrostatic attraction at physiological pH, thereby facilitating gene delivery. Many factors influence the gene transfection efficiency of cationic polymers, including their structure, molecular weight, and surface charge. Outstanding representatives of polymers that have emerged over the last decade to be used in gene therapy are synthetic polymers such as poly(l-lysine), poly(l-ornithine), linear and branched polyethyleneimine, diethylaminoethyl-dextran, poly(amidoamine) dendrimers, and poly(dimethylaminoethyl methacrylate). Natural polymers, such as chitosan, dextran, gelatin, pullulan, and synthetic analogs, with sophisticated features like guanidinylated bio-reducible polymers were also explored. This review outlines the introduction of polymers in medicine, discusses the methods of polymer synthesis, addressing top down and bottom up techniques. Evaluation of functionalization strategies for therapeutic and formulation stability are also highlighted. The overview of the properties, challenges, and functionalization approaches and, finally, the applications of the polymeric delivery systems in gene therapy marks this review as a unique one-stop summary of developments in this field.

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Scaffold-Mediated Gene Delivery for Osteochondral Repair

Pharmaceutics ◽

10.3390/pharmaceutics12100930 ◽

2020 ◽

Vol 12 (10) ◽

pp. 930 ◽

Cited By ~ 1

Author(s):

Henning Madry ◽

Jagadeesh Kumar Venkatesan ◽

Natalia Carballo-Pedrares ◽

Ana Rey-Rico ◽

Magali Cucchiarini

Keyword(s):

Gene Delivery ◽

Viral Vectors ◽

Viral Gene ◽

Therapeutic Gene ◽

Therapeutic Concept ◽

Osteochondral Repair ◽

Gene Vectors ◽

A Site

Osteochondral defects involve both the articular cartilage and the underlying subchondral bone. If left untreated, they may lead to osteoarthritis. Advanced biomaterial-guided delivery of gene vectors has recently emerged as an attractive therapeutic concept for osteochondral repair. The goal of this review is to provide an overview of the variety of biomaterials employed as nonviral or viral gene carriers for osteochondral repair approaches both in vitro and in vivo, including hydrogels, solid scaffolds, and hybrid materials. The data show that a site-specific delivery of therapeutic gene vectors in the context of acellular or cellular strategies allows for a spatial and temporal control of osteochondral neotissue composition in vitro. In vivo, implantation of acellular hydrogels loaded with nonviral or viral vectors has been reported to significantly improve osteochondral repair in translational defect models. These advances support the concept of scaffold-mediated gene delivery for osteochondral repair.

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