Abstract
Immune responses to expressed foreign transgenes continue to hamper progress of gene therapy development. Translated foreign proteins with intracellular location are generally less accessible to the immune system, nevertheless they can be presented to the immune system through both MHC Class I and Class II pathways. When the foreign protein luciferase was expressed following intramuscular delivery of plasmid DNA in outbred mice, expression rapidly declined over 4 weeks. Through modifications to the expression plasmid and the luciferase transgene we examined the effect of detargeting expression away from antigen-presenting cells (APCs), targeting expression to skeletal muscle and fusion with glycine-alanine repeats (GAr) that block MHC-Class I presentation on the duration of luciferase expression. De-targeting expression from APCs with miR142-3p target sequences incorporated into the luciferase 3’UTR reduced the humoral immune response to both native and luciferase modified with a short GAr sequence but did not prolong the duration of expression. When a skeletal muscle specific promoter was combined with the miR target sequences the humoral immune response was dampened and luciferase expression persisted at higher levels for longer. Interestingly, fusion of luciferase with a longer GAr sequence promoted the decline in luciferase expression and increased the humoral immune response to luciferase. These studies demonstrate that expression elements and transgene modifications can alter the duration of transgene expression but other factors will need to overcome before foreign transgenes expressed in skeletal muscle are immunologically silent.
Keywords: Gene therapy, luciferase, microRNA, plasmid DNA, skeletal muscle, tissue-specific promoter, transgene immunogenicity.
Current Gene Therapy
Title:Effects of APC De-Targeting and GAr Modification on the Duration of Luciferase Expression from Plasmid DNA Delivered to Skeletal Muscle
Volume: 15 Issue: 1
Author(s): Maria C. Subang, Rewas Fatah, Ying Wu, Drew Hannaman, Jason Rice, Claire F. Evans, Yuti Chernajovsky and David Gould
Affiliation:
Keywords: Gene therapy, luciferase, microRNA, plasmid DNA, skeletal muscle, tissue-specific promoter, transgene immunogenicity.
Abstract: Immune responses to expressed foreign transgenes continue to hamper progress of gene therapy development. Translated foreign proteins with intracellular location are generally less accessible to the immune system, nevertheless they can be presented to the immune system through both MHC Class I and Class II pathways. When the foreign protein luciferase was expressed following intramuscular delivery of plasmid DNA in outbred mice, expression rapidly declined over 4 weeks. Through modifications to the expression plasmid and the luciferase transgene we examined the effect of detargeting expression away from antigen-presenting cells (APCs), targeting expression to skeletal muscle and fusion with glycine-alanine repeats (GAr) that block MHC-Class I presentation on the duration of luciferase expression. De-targeting expression from APCs with miR142-3p target sequences incorporated into the luciferase 3’UTR reduced the humoral immune response to both native and luciferase modified with a short GAr sequence but did not prolong the duration of expression. When a skeletal muscle specific promoter was combined with the miR target sequences the humoral immune response was dampened and luciferase expression persisted at higher levels for longer. Interestingly, fusion of luciferase with a longer GAr sequence promoted the decline in luciferase expression and increased the humoral immune response to luciferase. These studies demonstrate that expression elements and transgene modifications can alter the duration of transgene expression but other factors will need to overcome before foreign transgenes expressed in skeletal muscle are immunologically silent.
Export Options
About this article
Cite this article as:
Subang C. Maria, Fatah Rewas, Wu Ying, Hannaman Drew, Rice Jason, Evans F. Claire, Chernajovsky Yuti and Gould David, Effects of APC De-Targeting and GAr Modification on the Duration of Luciferase Expression from Plasmid DNA Delivered to Skeletal Muscle, Current Gene Therapy 2015; 15 (1) . https://dx.doi.org/10.2174/1566523214666141114204943
DOI https://dx.doi.org/10.2174/1566523214666141114204943 |
Print ISSN 1566-5232 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5631 |
Call for Papers in Thematic Issues
Programmed Cell Death Genes in Oncology: Pioneering Therapeutic and Diagnostic Frontiers (BMS-CGT-2024-HT-45)
Programmed Cell Death (PCD) is recognized as a pivotal biological mechanism with far-reaching effects in the realm of cancer therapy. This complex process encompasses a variety of cell death modalities, including apoptosis, autophagic cell death, pyroptosis, and ferroptosis, each of which contributes to the intricate landscape of cancer development and ...read more

- Author Guidelines
- Bentham Author Support Services (BASS)
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Anti-Infective Strategies of the Future: Is there Room for Species-Specific Antibacterial Agents?
Current Pharmaceutical Design Involvement of the Transforming Growth Factor β in the Pathogenesis of Hereditary Hemorrhagic Telangiectasia
Current Pharmaceutical Design Novel Insights for Multiple Sclerosis and Demyelinating Disorders with Apoptosis, Autophagy, FoxO, and mTOR
Current Neurovascular Research Editorial (Thematic Issue: Nanotechnology for Drug Delivery Applications)
Current Drug Delivery Development ofNovel Compounds to Treat Autoimmune and Inflammatory Diseases and Graft Versus Host Reactions
Endocrine, Metabolic & Immune Disorders - Drug Targets Treatment of Diabetic Foot Ulcer: An Overview Strategies for Clinical Approach
Current Diabetes Reviews Subject Index To Volume 2
Current Rheumatology Reviews Cannabinoids as Therapeutic Agents for Ablating Neuroinflammatory Disease
Endocrine, Metabolic & Immune Disorders - Drug Targets Tumor Necrosis Factor Inhibitors as Therapeutic Choice in Psoriasis
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Review of Pediatric Osteoarticular Infections
Reviews on Recent Clinical Trials Congenital Abnormalities: Consequence of Maternal Zika Virus Infection: A Narrative Review
Infectious Disorders - Drug Targets The Role of Microglial Cells on Neuroinflammation: Possible Therapeutic Applications
Recent Patents on Regenerative Medicine Molecularly Imprinted Sol-Gel Materials for Medical Applications
Current Topics in Medicinal Chemistry Instructions from the Vascular System - Directing Neural Stem Cell Fate in Health and Disease
Current Medicinal Chemistry Monoclonal Antibodies in the Treatment of Multiple Sclerosis
Current Medicinal Chemistry Regulatory T Cells in Central Nervous System: in Health and Disease
Central Nervous System Agents in Medicinal Chemistry Anti-Inflammatory Therapeutic Approaches to Reduce Acute Atherosclerotic Complications
Current Pharmaceutical Biotechnology Neurovirulence of SARS CoV2: From Clinical Data to Preclinical Neuropsychological Exploration
Coronaviruses Diagnostic Value of 99m Tc- Labeled-Ubiquicidin 29-41 (99m Tc-UBI) Scan in the Diagnosis of Vertebral Osteomyelitis
Current Medical Imaging Neurobrucellosis: A Case Report with an Unusual Presentation
Recent Advances in Anti-Infective Drug Discovery