Pinyon Jeremy L, von Jonquieres Georg, Mow Stephen L, Abed Amr Al, Lai Keng-Yin, Manoharan Mathumathi, Crawford Edward N, Xue Stanley H, Smith-Moore Sarah, Caproni Lisa J, Milsom Sarah, Klugmann Matthias, Lovell Nigel H, Housley Gary D
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.
Charles Perkins Centre, School of Medical Sciences, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW, 2006, Australia.
Adv Sci (Weinh). 2025 Jan;12(3):e2406545. doi: 10.1002/advs.202406545. Epub 2024 Nov 27.
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. Proof-of-concept studies for "single capacitive discharge conductivity-clamped gene electrotransfer" (SCD-CC-GET) deep tissue delivery of naked DNA and mRNA in the mouse hindlimb skeletal muscle achieve stable (>18 month) expression of luciferase reporter synthetic DNA, and mRNA encoding the reporter yield rapid onset (<3 h) high transient expression for several weeks. Delivery of DNAs encoding secreted alkaline phosphatase and Cal/09 influenza virus hemagglutinin antigen generate high systemic circulating recombinant protein levels and antibody titres. The findings support adoption of SCD-CC-GET for vaccines and immunotherapies, and extend the utility of this technology to meet the demand for efficient vector-free, precision, deep tissue delivery of NA therapeutics.
基于病毒载体和脂质纳米颗粒的基因递送在时空控制、转基因包装大小和载体免疫反应性方面存在局限性，这影响了核酸（NA）疗法的转化应用。在新兴的DNA尤其是基于RNA的基因治疗领域，无载体递送平台被视为一项关键的未满足需求。在此，本研究通过使用单针外形因素对“裸露的”聚阴离子DNA/ mRNA进行基因电穿孔（GET）来应对这些挑战，该外形因素支持基于“电透镜”的局部电场压缩以及组织电导率的局部控制，从而实现单次电容放电的最小电荷基因递送。在小鼠后肢骨骼肌中对裸露DNA和mRNA进行“单次电容放电电导率钳制基因电穿孔”（SCD-CC-GET）深层组织递送的概念验证研究表明，荧光素酶报告基因合成DNA实现了稳定（> 18个月）表达，编码报告基因的mRNA在数周内产生快速起效（<3小时）的高瞬时表达。递送编码分泌碱性磷酸酶和Cal/09流感病毒血凝素抗原的DNA可产生高全身性循环重组蛋白水平和抗体滴度。这些发现支持将SCD-CC-GET用于疫苗和免疫疗法，并扩展了该技术的用途，以满足对高效、无载体、精准的NA疗法深层组织递送的需求。
