Sales Conniff Amanda, Tur Jared, Kohena Kristopher, Zhang Min, Gibbons Justin, Heller Loree C
Department of Medical Engineering, University of South Florida, Tampa, Florida, USA.
USF Genomics Core, University of South Florida, Tampa, Florida, USA.
Bioelectricity. 2024 Jun 12;6(2):80-90. doi: 10.1089/bioe.2022.0041. eCollection 2024 Jun.
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.
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.
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.
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.
诸如骨骼肌等组织已成为编码疫苗和治疗药物的质粒DNA（pDNA）递送的目标。电脉冲（电穿孔或电转染）的应用可增加细胞膜通透性，以增强质粒递送和表达。然而，DNA电转染对肌肉组织的分子效应却鲜有描述。
在肌肉内进行质粒电转染4小时后，我们通过RNA测序评估基因表达变化。通过基因本体（GO）通路富集分析对差异表达基因进行分析。
GO分析突出了许多富集的分子功能。由脉冲施加所调控的术语与肌肉应激、细胞骨架和炎症有关。由pDNA注射所调控的术语与DNA定向反应及其控制有关。由pDNA电转染所调控的几个术语与由脉冲施加所调控的术语相似。然而，与pDNA注射相关的术语有所不同，重点在于质粒进入细胞和细胞内运输。
每种肌肉刺激都会导致特定的调控分子功能。识别由肌肉内DNA电转染驱动的独特内在分子变化将有助于预防性和治疗性基因疗法的设计。
