Background: Tissues, such as skeletal muscle, have been targeted for the delivery of plasmid DNA (pDNA) encoding vaccines and therapeutics. The application of electric pulses (electroporation or electrotransfer) increases cell membrane permeability to enhance plasmid delivery and expression. However, the molecular effects of DNA electrotransfer on the muscle tissue are poorly characterized.
Materials And Methods: Four hours after intramuscular plasmid electrotransfer, we evaluated gene expression changes by RNA sequencing. Differentially expressed genes were analyzed by gene ontology (GO) pathway enrichment analysis.
Results: GO analysis highlighted many enriched molecular functions. The terms regulated by pulse application were related to muscle stress, the cytoskeleton and inflammation. The terms regulated by pDNA injection were related to a DNA-directed response and its control. Several terms regulated by pDNA electrotransfer were similar to those regulated by pulse application. However, the terms related to pDNA injection differed, focusing on entry of the plasmid into the cells and intracellular trafficking.
Conclusion: Each muscle stimulus resulted in specific regulated molecular functions. Identifying the unique intrinsic molecular changes driven by intramuscular DNA electrotransfer will aid in the design of preventative and therapeutic gene therapies.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11304878 | PMC |
| http://dx.doi.org/10.1089/bioe.2022.0041 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Phase I trial of phIL12 plasmid intratumoral gene electrotransfer in patients with basal cell carcinoma in head and neck region.
Eur J Surg Oncol
January 2025
Institute of Oncology Ljubljana, Slovenia; Faculty of Health Sciences, University of Ljubljana, Slovenia. Electronic address:
Introduction: In the treatment of cancer, immunomodulatory approaches are developed to support the organism in fighting cancer or to enhance the immunomodulatory effects of local ablative techniques. To this end, we conducted an interventional, open-label, single-arm Phase I trial to evaluate the safety and tolerability of intratumoral phIL12 plasmid DNA gene electrotransfer as primary objectives.
Methods: The study was dose-escalating with 3 consecutive cohorts of 3 patients per phIL12 dose level (0.
Vector-Free Deep Tissue Targeting of DNA/RNA Therapeutics via Single Capacitive Discharge Conductivity-Clamped Gene Electrotransfer.
Adv Sci (Weinh)
January 2025
Translational Neuroscience Facility, Department of Physiology, School of Biomedical Sciences, Graduate School of Biomedical Engineering, Tyree Institute for Health Engineering (IHealthE), UNSW, Sydney, NSW, 2052, Australia.
Viral vector and lipid nanoparticle based gene delivery have limitations around spatiotemporal control, transgene packaging size, and vector immune reactivity, compromising translation of nucleic acid (NA) therapeutics. In the emerging field of DNA and particularly RNA-based gene therapies, vector-free delivery platforms are identified as a key unmet need. Here, this work addresses these challenges through gene electrotransfer (GET) of "naked" polyanionic DNA/mRNA using a single needle form-factor which supports "electro-lens" based compression of the local electric field, and local control of tissue conductivity, enabling single capacitive discharge minimal charge gene delivery.
View Article and Find Full Text PDFRNA Blotting by Electrotransfer and RNA Detection.
Methods Mol Biol
November 2024
Institut de Biologie Physico-Chimique, UMR8226, CNRS, Sorbonne Université, Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Paris, France.
Northern blot, or RNA blot, is a widely used technique in molecular biology to detect and semi-quantify specific RNAs. The advantage of this method is its ability to simultaneously estimate the sizes and quantities of degraded or processed RNA products. Northern blotting involves the use of electrophoresis to separate RNA samples by size.
View Article and Find Full Text PDFNon-clinical evaluation of pmIL12 gene therapy for approval of the phase I clinical study.
Sci Rep
September 2024
Institute of Oncology Ljubljana, 1000, Ljubljana, Slovenia.
Immunotherapeutic drugs are promising medicines for cancer treatment. A potential candidate for immunotherapy is interleukin-12 (IL-12), a cytokine well known for its ability to mediate antitumor activity. We developed a plasmid encoding human IL-12 devoid of an antibiotic resistance gene (phIL12).
View Article and Find Full Text PDFIL-12 and PD-1 peptide combination gene therapy for the treatment of melanoma.
Mol Ther Nucleic Acids
September 2024
Department of Medical Engineering, University of South Florida, Morsani College of Medicine and College of Engineering, 12901 Bruce B. Downs Blvd., MDC111, Tampa, FL 33612, USA.
Interleukin-12 (IL-12) gene electrotransfer (GET) delivery is highly effective in inducing long-term, complete regression in mouse and human melanoma and other solid tumors. Therapeutic efficacy is enhanced by immune checkpoint inhibitors, and the combination of IL-12 plasmid GET (pIL-12 GET) and anti-programmed cell death protein 1 (PD-1) monoclonal antibodies has reached clinical trials. In this study, we designed peptides and plasmids encoding the mouse homologs of the pembrolizumab and nivolumab programmed cell death 1 ligand 1 (PD-L1) binding regions.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!