Gene Therapy Engineering Research Articles

Nanotechnology and CRISPR/Cas-Mediated Gene Therapy Strategies: Potential Role for Treating Genetic Disorders.

Gene therapy has made substantial progress in the treatment of the genetic diseases, focussing on the reduction of characteristics of recessive/dominant disorders, as well as various cancers. Extensive research has been conducted in the past few decades to investigate the application of nanotechnology and CRISPR/Cas technology in gene therapy. Nanotechnology due to attributes such has targeted drug delivery, controlled release, scalability and low toxicity has gained attention of the medical world. CRISPR/Cas9 system is considered as an impactful genome editing tool in the area of next-generation therapeutics and molecular diagnostics. CRISPR technology emphasises on gene editing, gene regulation modulation, and formulation of defined genetic changes. Its applications in treatment of the genetic disorders are extended beyond traditional therapies. These techniques are being explored as treatment of several genetic disorders including Duchenne muscular dystrophy, cystic fibrosis, Alzheimer's disease, Parkinson's disease, and Huntington disease. Despite considerable therapeutic potential of gene therapy, several obstacles must be addressed before it can be widely adopted in clinical practice, particularly in terms of ensuring safety and effectiveness. As research advances in this captivating field, these therapies will become the primary treatments and will have significant beneficial effects on the lives of patients with genetic disorders.

Recent advances and current status of gene therapy for epilepsy.

Epilepsy is a common neurological disorder with complex pathogenic mechanisms, and refractory epilepsy often lacks effective treatments. Gene therapy is a promising therapeutic option, with various preclinical experiments achieving positive results, some of which have progressed to clinical studies. This narrative review was conducted by searching for papers published in PubMed/MEDLINE with the following single and/or combination keywords: epilepsy, children, neurodevelopmental disorders, genetics, gene therapy, vectors, transgenes, receptors, ion channels, micro RNAs (miRNAs), clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas)9 (CRISPR/Cas9), expression regulation, optogenetics, chemical genetics, mitochondrial epilepsy, challenges, ethics, and disease models. Currently, gene therapy research in epilepsy primarily focuses on symptoms attenuationmediated by viral vectors such as adeno-associated virus and other types. Advances in gene therapy technologies, such as CRISPR/Cas9, have provided a new direction for epilepsy treatment. However, the clinical application still faces several challenges, including issues related to vectors, models, expression controllability, and ethical considerations. Here, we summarize the relevant research and clinical advances in gene therapy for epilepsy and outline the challenges facing its clinical application. In addition to the shortcomings inherent in gene therapy components, the reconfiguration of excitatory and inhibitory properties in epilepsy treatment is a delicate process. On-demand, cell-autonomous treatments and multidisciplinary collaborations may be crucial in addressing these issues. Understanding gene therapy for epilepsy will help clinicians gain a clearer perception of the research progress and challenges, guiding the design of future clinical protocols and research decisions.

Prospects for gene therapy in polycystic kidney disease.

We aim to provide an updated perspective on the recent advancements in gene therapy for polycystic kidney disease (PKD), a genetic disorder with significant morbidity. Given the rapid evolution of gene therapy technologies and their potential for treating inherited diseases, this review explores the therapeutic prospects and challenges in applying these technologies to PKD. Significant progress has been made in understanding the genetic underpinnings of PKD, making it a prime candidate for gene therapy. Re-expression of the PKD genes, treatment with the C-terminal tail of polycystin 1 protein and antagomir therapy against miR-17 have shown promise in reducing cyst formation and preserving kidney function. The rapid development of gene-editing tools, antisense oligonucleotide-based strategies, programmable RNA, and advanced gene delivery systems has opened new possibilities for PKD treatment. However, challenges such as off-target effects, delivery efficiency, and long-term safety remain significant barriers to clinical application. Current research highlights the transformative potential of gene therapy for PKD. Ongoing studies are crucial to overcoming existing challenges and translating these findings into clinical practice. We highlight the need for multidisciplinary efforts to optimize gene-editing technologies and ensure their safety and efficacy in treating PKD.

NF-κB-activated oncogene inhibition strategy for cancer gene therapy.

NF-κB is a promising target for cancer treatment because of its overactivation in almost all cancers but countless NF-κB inhibitors rarely became clinical drugs due to side effects. In contrast to traditional cancer treatments aimed at inhibiting NF-κB activity, this study develop a novel approach termed HOPE, which focuses on activating the exogenous effector gene CRISPR-Cas13a within cancer cells, achieved by utilizing the NF-κB-specific promoter DMP previously constructed, then targets and suppresses the expression of oncogenes TERT, PLK1, KRAS and MYC at mRNA level. We evaluated the antitumour effects of HOPE in various cultured cells and confirmed it could induce obvious the death of cancer cells without affecting normal cells. By packaging HOPE into adeno-associated virus (AAV) and intravenously injected it to treat mice that were subcutaneously transplanted with colorectal cancer. This validated that rAAV-HOPE could significantly inhibit tumour growth without side effects. Based on the scRNA-seq data, we observed that HOPE could activate the immune system and decrease the proportion of cancer cells, particularly reducing the stemness of cancer cells. This study elucidates an important role of HOPE in inhibiting cancer cell growth both in vitro and in vivo, additionally provides a novel therapeutic technology for cancer gene therapy.

COVID-19 Modified mRNA “Vaccines”: Lessons Learned from Clinical Trials, Mass Vaccination, and the Bio-Pharmaceutical Complex, Part 2

The COVID-19 modified mRNA (modmRNA) lipid nanoparticle-based “vaccines” are not classical antigen-based vaccines but instead prodrugs informed by gene therapy technology. Of considerable note, these products have been linked to atypical adverse and serious adverse event profiles. As discussed in Part 1, health-related risks and drawbacks were drastically misreported and underreported in the Pfizer and Moderna trial evaluations of these genetic products. Now in Part 2, we examine the main structural and functional aspects of these injectables. The COVID-19 modmRNA injectable products introduce a unique set of biological challenges to the human body with the potential to induce an extensive range of adverse, crippling, and life-threatening effects. Based on the fact that there is no current method to quantify host (cell-based) spike protein production in vivo following injection with these prodrugs, there is no standard “dose”. This is in part due to differences in spike protein production output, which depends on cell metabolism and transfection efficiency. It is therefore difficult to predict adverse event profiles on an individual basis, but considering that millions of adults across the world have reported severe and serious adverse events in the context of these modmRNA COVID-19 products, valid concerns are raised regarding injection of infants and younger age groups for whom COVID-19 poses only minimal risks. We address the process-related genetic impurities inherent in mass production of these products, and the potential risks posed by these contaminants. We then categorize the principal adverse events associated with the modmRNA products with a brief systems-based synopsis of each of the six domains of potential harms: (1) cardiovascular, (2) neurological, (3) hematologic; (4) immunological, (5) oncological, and (6) reproductive. We conclude with a discussion of the primary public health and regulatory issues arising from this evidence-informed synthesis of the literature and reiterate the urgency of imposing a global moratorium on the modmRNA-LNP-based platform.

Cardiac Arrhythmias and Their Management: An In-Depth Review of Current Practices and Emerging Therapies.

Cardiac arrhythmias encompass a range of conditions characterized by abnormal heart rhythms, affecting millions globally and significantly contributing to morbidity and mortality. This review provides a comprehensive analysis of the current practices and emerging therapies in managing cardiac arrhythmias, covering their definition, classification, epidemiology, and the critical importance of effective management. It explores the pathophysiology underlying various arrhythmias, including the mechanisms of arrhythmogenesis, such as re-entry, automaticity, and triggered activity. The review details the latest diagnostic tools, including ECG, Holter monitoring, and electrophysiological studies, and discusses the clinical presentation of different arrhythmias, from supraventricular to ventricular types and bradyarrhythmias. We examine current pharmacological and non-pharmacological treatment strategies, such as antiarrhythmic drugs, catheter ablation, and device therapy, highlighting their efficacy and limitations. Furthermore, the review delves into emerging therapies, including advanced catheter ablation techniques, novel antiarrhythmic agents, gene therapy, and innovative device technologies like leadless pacemakers and subcutaneous implantable cardioverter-defibrillators (ICDs). Special considerations for managing arrhythmias in diverse populations, including pediatrics, the elderly, and pregnant women, are discussed. Additionally, the review explores future directions in arrhythmia management, emphasizing personalized medicine, artificial intelligence applications, and the integration of advanced technologies in diagnosis and treatment. By synthesizing current knowledge and prospects, this review aims to enhance understanding and promote advancements in the field, ultimately improving patient outcomes with cardiac arrhythmias.

Targeted gene therapy for rare genetic kidney diseases

Chronic kidney disease (CKD) is one of the leading causes of mortality worldwide because of kidney failure and the associated challenges of its treatment including dialysis and kidney transplantation. About one-third of CKD cases are linked to inherited monogenic factors, making them suitable for potential gene therapy interventions. However, the intricate anatomical structure of the kidney poses a challenge, limiting the effectiveness of targeted gene delivery to the renal system. In this review, we explore the progress made in the field of targeted gene therapy approaches and their implications for rare genetic kidney disorders, examining preclinical studies and prospects for clinical application. In vivo gene therapy is most commonly used for kidney-targeted gene delivery and involves administering viral and non-viral vectors through various routes such as systemic, renal vein and renal arterial injections. Small nucleic acids have also been used in preclinical and clinical studies for treating certain kidney disorders. Unexpectedly, hematopoietic stem and progenitor cells have been used as an ex vivo gene therapy vehicle for kidney gene delivery, highlighting their ability to differentiate into macrophages within the kidney, forming tunneling nanotubes that can deliver genetic material and organelles to adjacent kidney cells, even across the basement membrane to target the proximal tubular cells. As gene therapy technologies continue to advance and our understanding of kidney biology deepens, there is hope for patients with genetic kidney disorders to eventually avoid kidney transplantation.

Advancements in Lipid Nanoparticle Technologies for Precision Targeting in Therapeutic Delivery

<img src=” https://s3.amazonaws.com/production.scholastica/article/121633/large/prnano_1412024bg2.jpeg?1721676909”> Lipid nanoparticles (LNPs) are at the forefront of targeted therapeutic delivery, serving as an innovative platform for precisely delivering therapeutic agents directly to specific cells or tissues. Therapeutic delivery seeks to enhance drug efficacy and safety by controlling the distribution of drugs within the body, thereby minimizing off-target effects while maximizing therapeutic impact. This review delves into advancements and innovations in LNP design to achieve unprecedented targeting precision, surmount biological barriers, and diminish off-target interactions. It encom-passes exploring targeting strategies, including passive targeting that leverages the enhanced permeability and retention effect, active targeting via receptor-ligand interactions, and stimuli-responsive methodologies for the controlled release of therapeutics. With a focus on novel tar-geting techniques, including endogenous targeting and strategic de-targeting, to enhance LNP de-livery specificity and therapeutic efficacy. This review articulates the critical role of lipid com-position, nanoparticle characterization, and surface engineering in tailoring LNPs for therapeutic applications. Through evaluating the forefront of LNP technology in drug delivery and gene therapy, this review forecasts the trajectory of nanoparticle-based therapeutics, envisioning their pivotal role in the advent of personalized medicine, thereby providing a foundation for future explorations aimed at refining and optimizing LNP systems for clinical applications.

Advancements in Gene Therapy and Stem Cell Applications in Interventional Cardiology: A Comprehensive Review

Gene therapy and stem cell applications represent the forefront of innovative treatments in interventional cardiology, offering potential solutions for previously intractable cardiovascular conditions. This review explores the current advancements, mechanisms, and clinical implications of gene therapy and stem cell technologies in the treatment of ischemic heart disease, heart failure, and myocardial infarction. The integration of these modalities into interventional procedures has the potential to revolutionize patient outcomes by promoting myocardial repair, enhancing angiogenesis, and improving cardiac function. Despite promising preclinical and early clinical results, challenges such as delivery methods, immunogenicity, and long-term efficacy remain. This article provides a detailed analysis of the latest research, clinical trials, and future directions in the field, highlighting the transformative potential and obstacles that need to be addressed for widespread clinical adoption.

Somatic Gene Therapy in the Prevention of Toxic Effects of Organophosphate Agents

Medical intervention in poisoning by organophosphate toxic agents (OPA) using atropine sulfate, 2-pyridinaldoxymethyl chloride (2-PAM), diazepam and other similar drugs can prevent the fatal outcome of poisoning. These drugs do not protect in case of sudden chemical attack and against post-exposure complications associated with permanent brain damage. The U.S. Department of Defense is funding research that can significantly simplify the protection of military personnel from OPA damage in the future. Their essence is in the use of gene therapy technologies, which allow experimental animals to produce their own proteins that destroy OPA and provide them with protection for several months. The aim of the work is to identify the achieved level of knowledge in the research using gene therapy technologies to create living objects resistant to OPA. The research method is analytical. The source base of the research are publications in scientific journals and descriptions of patents. Discussion of the results. As an enzyme that breaks down OPA in such experiments, genetically modified paraoxanase 1 (PON1) showed the greatest efficiency. PON1 hydrolyzes G-type OPAs, paraoxone, chlorpyrifosoxone, diazoxone and several other organophosphates. Adenoassociated virus vectors (AAV8, etc.) were used to introduce the gene encoding PON1 into the animal's body. A single injection of AAV8 carrying the recombinant PON1-IF11 gene (AAV8-PON1-IF11) resulted in high expression and secretion of the recombinant PON1-IF11 protein into the bloodstream and provided asymptomatic protection against multiple lethal doses of G-type OPA for at least 5 months. These studies are still in their early stage. An analysis of the affiliation of the authors of publications and patents showed a high involvement of the U.S. military department and its cooperating organizations (DTRA, etc.) in such research. Conclusion. Given the fascination in the West with the ideas of human modification using gene therapy methods, this direction will be intensively developed for military purposes. At the same time, the idea of pre-created resistance to OPA is in demand by the widespread use of organophosphates in agriculture. The author believes that it would be safer to use allogeneic mesenchymal stem cells transfected with genetically modified PON1 variants with enhanced enzyme activity. This resistance to OP agents can be health protective and lifesaving in soldiers in real combat when the enemy uses these agents. However, this approach must be based on a strong experimental background. The door is open, the technologies are available.

Detection Method for Gene Doping in a Mouse Model Expressing Human Erythropoietin from Adeno-Associated Virus Vector-9.

With the rapid development of gene therapy technology in recent years, its abuse as a method of sports doping in athletics has become a concern. However, there is still room for improvement in gene-doping testing methods, and a robust animal model needs to be developed. Therefore, the purposes of this study were to establish a model of gene doping using recombinant adeno-associated virus vector-9, including the human erythropoietin gene (rAAV9-hEPO), and to establish a relevant testing method. First, it was attempted to establish the model using rAAV9-hEPO on mice. The results showed a significant increase in erythrocyte volume accompanied by an increase in spleen weight, confirming the validity of the model. Next, we attempted to detect proof of gene doping by targeting DNA and RNA. Direct proof of gene doping was detected using a TaqMan-qPCR assay with certain primers/probes. In addition, some indirect proof was identified in RNAs through the combination of a TB Green qPCR assay with RNA sequencing. Taken together, these results could provide the foundation for an effective test for gene doping in human athletes in the future.

Application and perspective of CRISPR/Cas9 genome editing technology in human diseases modeling and gene therapy.

The Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) mediated Cas9 nuclease system has been extensively used for genome editing and gene modification in eukaryotic cells. CRISPR/Cas9 technology holds great potential for various applications, including the correction of genetic defects or mutations within the human genome. The application of CRISPR/Cas9 genome editing system in human disease research is anticipated to solve a multitude of intricate molecular biology challenges encountered in life science research. Here, we review the fundamental principles underlying CRISPR/Cas9 technology and its recent application in neurodegenerative diseases, cardiovascular diseases, autoimmune related diseases, and cancer, focusing on the disease modeling and gene therapy potential of CRISPR/Cas9 in these diseases. Finally, we provide an overview of the limitations and future prospects associated with employing CRISPR/Cas9 technology for diseases study and treatment.

Adenine base editor-based correction of the cardiac pathogenic Lmna c.1621C > T mutation in murine hearts.

Base editors are emerging as powerful tools to correct single-nucleotide variants and treat genetic diseases. In particular, the adenine base editors (ABEs) exhibit robust and accurate adenine-to-guanidine editing capacity and have entered the clinical stage for cardiovascular therapy. Despite the tremendous progress using ABEs to treat heart diseases, a standard technical route toward successful ABE-based therapy remains to be fully established. In this study, we harnessed adeno-associated virus (AAV) and a mouse model carrying the cardiomyopathy-causing Lmna c.1621C > T mutation to demonstrate key steps and concerns in designing a cardiac ABE experiment invivo. We found DeepABE as a reliable deep-learning-based model to predict ABE editing outcomes in the heart. Screening of sgRNAs for a Cas9 mutant with relieved protospacer adjacent motif (PAM) allowed the reduction of bystander editing. The ABE editing efficiency can be significantly enhanced by modifying the TadA and Cas9 variants, which are core components of ABEs. The ABE systems can be delivered into the heart via either dual AAV or all-in-one AAV vectors. Together, this study showcased crucial technical considerations in designing an ABE system for the heart and pointed out major challenges in further improvement of this new technology for gene therapy.

BONE TISSUE REGENERATION INDUCED BY LOCAL DELIVERY OF BONE MORPHOGENETIC PROTEIN 2 FROM PRO-ANGIOGENIC NEAR-INFRARED-RESPONSIVE HYDROGELS

In this work, we combined tissue engineering and gene therapy technologies to develop a therapeutic platform for bone regeneration. We have developed photothermal fibrin-based hydrogels that incorporate degradable CuS nanoparticles (CuSNP) which transduce incident near-infrared (NIR) light into heat. A heat-activated and rapamycin-dependent transgene expression system was incorporated into mesenchymal stem cells to conditionally control the production of bone morphogenetic protein 2 (BMP-2). Genetically engineered cells were entrapped in the photothermal hydrogels. In the presence of rapamycin, photoinduced mild hyperthermia induced the release of BMP-2 from the NIR responsive cell constructs. Transcriptome analysis of BMP-2 expressing cells showed a signature of induced genes related to stem cell proliferation and angiogenesis. We next generated 4 mm diameter calvarial defects in the left parietal bone of immunocompetent mice. The defects were filled with NIR-responsive hydrogels entrapping cells that expressed BMP-2 under the control of the gene circuit. After one and eight days, rapamycin was administered intraperitoneally followed by irradiation with an NIR laser. Ten weeks after implantation, the animals were euthanized and samples from the bone defect zone were processed for histological analysis using Masson's trichrome staining and for immunohistochemistry analyses using specific CD31 and CD105 antibodies. Samples from mice that were only administered rapamycin or vehicle or that were only NIR-irradiated showed the persistence of fibrous tissue bridging the defect. In animals that were treated with rapamycin, NIR irradiation of implants resulted in the formation of new mineralized tissue with a high degree of vascularization, thus indicating the therapeutic potential of the approach.Acknowledgements: This research was supported by grants RTI2018-095159-B-I00 and PID2021-126325OB-I00 (MCIN/AEI/10.13039/501100011033 and “ERDF A way of making Europe”), by grant P2022/BMD- 7406 (Regional Government of Madrid). M.A.L-J. is the recipient of predoctoral fellowship PRE2019-090430 (MCIN/AEI/10.13039/501100011033).

Research on the gene therapy and prevention of lung cancer

In recent years, there have been significant advancements in molecular biology, gene therapy technology, and high-tech testing techniques. As our understanding of the pathogenesis of lung cancer continues to deepen, it has become evident that gene therapy plays a crucial role in its treatment. This article provides an overview of tumor cell development, various gene therapy techniques including suicide gene therapy, silent gene therapy, cancer suppressor genetic therapy, and antibody genetic therapy. Additionally, it analyzes the application value, treatment technology, and safety concerns associated with gene therapy. The discussion also covers the safety issues and current application status of viral and nonviral vectors in gene therapy, comparing the advantages and disadvantages of different vector types. Furthermore, the article summarizes the future directions for designing and improving gene delivery vectors, along with their clinical application prospects. Ultimately, this article offers insights into the significance and treatment approaches for early prevention of lung cancer.

A Finnish Survey of Adverse Effects of COVID-19 Injectables and the Functionality of the Medical System

The declaration of the COVID-19 pandemic brought unprecedented containment measures across multiple countries, with the announced intent of saving lives. Among them was the rapid development of prophylactic vaccines, with the promise of no corners cut, but warning signals from safety monitoring systems around the world raised concerns. The current survey was undertaken to evaluate the Finnish medical system in light of adverse effects from COVID-19 vaccination. An online survey consisting of 96 questions (see the Appendix) was promoted through social media. Persons believing they had been injured by any COVID-19 injectable contacted the second author who first verified the identity of the respondent and then gave that person the key to complete the questionnaire. The open period lasted from 5 March 2023 to 25 July 2023.: Out of 67 respondents 63 completed the survey and the collected data was analyzed. Of 63 respondents, 55 (87%) were females and 8 (13%) males. The majority, 56 (89%) of them, were fully employed, and 29 (46%) were social sector or medical professionals. None were previously “vaccine hesitant”. All received from 1 to 4 doses of COVID-19 injectable(s), and 19 (30%) reported doing so under duress. Despite having complied with mandatory vaccination, 13 of the individuals (21%) lost their jobs, and 13 (21%) changed their profession because of the shots. Of the vaccine recipients, 40 (63%) reported that they were only partially able to continue their daily routines, and 17 (27%) reported not being able to maintain daily routines at all. Upon noticing adverse effects, 58 (92%) of the respondents opted to refuse any further doses, but 29 (46%) of them were compelled to take further shots despite their symptoms of prior vaccine injury. A broad range of system disorders was reported, including, notably, neurological and cardiovascular problems. Respondents were asked to describe their experience with the medical system and the healthcare workers they called on for help. Results show that COVID-19 injectables elicited a broad range of severe symptoms, raising grave concerns about the dangers of the products and casting doubt on whether promised benefits outweigh the risks that are increasingly well-known. Dozens of vaccinees in this data set were gravely impacted and these cases can only represent a small fraction of the larger number of injuries that have already occurred and continue to occur in Finland alone. Further use of these and future products based on this gene therapy technology should be suspended or discontinued altogether and medical doctors must educate and prepare themselves to face the consequences of this worldwide public health disaster. Open and sincere debates on the implications for medical ethics and the risk-benefit ratio of this novel technology are warranted.

A Finnish Survey of Adverse Effects of COVID-19 Injectables and the Functionality of the Medical System

The declaration of the COVID-19 pandemic brought unprecedented containment measures across multiple countries, with the announced intent of saving lives. Among them was the rapid development of prophylactic vaccines, with the promise of no corners cut, but warning signals from safety monitoring systems around the world raised concerns. The current survey was undertaken to evaluate the Finnish medical system in light of adverse effects from COVID-19 vaccination. An online survey consisting of 96 questions (see Appendix) was promoted through social media. Persons believing they had been injured by any COVID-19 injectable contacted the second author who first verified the identity of the respondent and then gave that person the key to complete the questionnaire. The open period lasted from 5 March 2023 to 25 July 2023.: Out of 67 respondents 63 completed the survey and the collected data was analyzed. Of 63 respondents, 55 (87%) were females and 8 (13%) males. The majority, 56 (89%) of them, were fully employed, and 29 (46%) were social sector or medical professionals. None were previously “vaccine hesitant”. All received from 1 to 4 doses of COVID-19 injectable(s), and 19 (30%) reported doing so under duress. Despite having complied with mandatory vaccination, 13 of the individuals (21%) lost their jobs, and 13 (21%) changed their profession because of the shots. Of the vaccine recipients, 40 (63%) reported that they were only partially able to continue their daily routines, and 17 (27%) reported not being able to maintain daily routines at all. Upon noticing adverse effects, 58 (92%) of the respondents opted to refuse any further doses, but 29 (46%) of them were compelled to take further shots despite their symptoms of prior vaccine injury. A broad range of system disorders was reported, including, notably, neurological and cardiovascular problems. Respondents were asked to describe their experience with the medical system and the healthcare workers they called on for help. Results show that COVID-19 injectables elicited a broad range of severe symptoms, raising grave concerns about the dangers of the products and casting doubt on whether promised benefits outweigh the risks that are increasingly well-known. Dozens of vaccinees in this data set were gravely impacted and these cases can only represent a small fraction of the larger number of injuries that have already occurred and continue to occur in Finland alone. Further use of these and future products based on this gene therapy technology should be suspended or discontinued altogether and medical doctors must educate and prepare themselves to face the consequences of this worldwide public health disaster. Open and sincere debates on the implications for medical ethics and the risk-benefit ratio of this novel technology are warranted.

Reversible, Covalent DNA Condensation Approach Using Chemical Linkers for Enhanced Gene Delivery.

Nonviral gene delivery has emerged as a promising technology for gene therapy. Nonetheless, these approaches often face challenges, primarily associated with lower efficiency, which can be attributed to the inefficient transportation of DNA into the nucleus. Here, we report a two-stage condensation approach to achieve efficient nuclear transport of DNA. First, we utilize chemical linkers to cross-link DNA plasmids via a reversible covalent bond to form smaller-sized bundled DNA (b-DNA). Then, we package the b-DNA into cationic vectors to further condense b-DNA and enable efficient gene delivery to the nucleus. We demonstrate clear improvements in the gene transfection efficiency in vitro, including with 11.6 kbp plasmids and in primary cultured neurons. Moreover, we also observed a remarkable improvement in lung-selective gene transfection efficiency in vivo by this two-stage condensation approach following intravenous administration. This reversible covalent assembly strategy demonstrates substantial value of nonviral gene delivery for clinical therapeutic applications.

Gene Therapy for Immunoglobulin E, Complement-Mediated, and Eosinophilic Disorders.

Immunoglobulin E, complement, and eosinophils play an important role in host defense, but dysfunction of each of these components can lead to a variety of human disorders. In this review, we summarize how investigators have adapted gene therapy and antisense technology to modulate immunoglobulin E, complement, and/or eosinophil levels to treat these disorders.

Glycogen storage disease type I: Genetic etiology, clinical manifestations, and conventional and gene therapies.

Glycogen storage disease type I (GSDI) is an inherited metabolic disorder characterized by a deficiency of enzymes or proteins involved in glycogenolysis and gluconeogenesis, resulting in excessive intracellular glycogen accumulation. While GSDI is classified into four different subtypes based on molecular genetic variants, GSDIa accounts for approximately 80%. GSDIa and GSDIb are autosomal recessive disorders caused by deficiencies in glucose-6-phosphatase (G6Pase-α) and glucose-6-phosphate-transporter (G6PT), respectively. For the past 50 years, the care of patients with GSDI has been improved following elaborate dietary managements. GSDI patients currently receive dietary therapies that enable patients to improve hypoglycemia and alleviate early symptomatic signs of the disease. However, dietary therapies have many limitations with a risk of calcium, vitamin D, and iron deficiency and cannot prevent long-term complications, such as progressive liver and renal failure. With the deepening understanding of the pathogenesis of GSDI and the development of gene therapy technology, there is great progress in the treatment of GSDI. Here, we review the underlying molecular genetics and the current clinical management strategies of GSDI patients with an emphasis on promising experimental gene therapies.