Mohan Ganesh, Khan Imran, Neumann Colby R, Jorge Miguel D, Ahmed Shahnur, Hulsman Luci, Sinha Mithun, Gordillo Gayle M, Sen Chandan K, Hassanein Aladdin H
Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA.
McGowan Institute for Regenerative Medicine, Department of Plastic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Mol Ther Nucleic Acids. 2024 Jan 18;35(1):102121. doi: 10.1016/j.omtn.2024.102121. eCollection 2024 Mar 12.
Lymphedema is chronic limb swelling resulting from lymphatic dysfunction. There is no cure for the disease. Clinically, a preventive surgical approach called immediate lymphatic reconstruction (ILR) has gained traction. Experimental gene-based therapeutic approaches (e.g., using viral vectors) have had limited translational applicability. Tissue nanotransfection (TNT) technology uses a direct, transcutaneous nonviral vector, gene delivery using a chip with nanochannel poration in response to a rapid (<100 ms) focused electric field. The purpose of this study was to experimentally prevent lymphedema using focal delivery of a specific gene (a master regulator of lymphangiogenesis). TNT was applied to the previously optimized lymphedematous mice tail (day 0) directly at the surgical site with genetic cargo loaded into the TNT reservoir: group I (sham) was given pCMV6 (expression vector backbone alone) and group II was treated with pCMV6-. Group II mice had decreased tail volume (47.8%) compared to sham and greater lymphatic clearance on lymphangiography. Immunohistochemistry showed greater lymphatic vessel density and RNA sequencing exhibited reduced inflammatory markers in group II compared to group I. prophylactically delivered using TNT to the surgical site on the day of injury decreased the manifestations of lymphedema in the murine tail model compared to control.
淋巴水肿是由淋巴功能障碍引起的慢性肢体肿胀。这种疾病无法治愈。临床上，一种名为即时淋巴重建（ILR）的预防性手术方法越来越受到关注。基于实验基因的治疗方法（例如使用病毒载体）在转化应用方面有限。组织纳米转染（TNT）技术使用一种直接的经皮非病毒载体，通过带有纳米通道穿孔的芯片在快速（<100毫秒）聚焦电场作用下进行基因递送。本研究的目的是通过局部递送特定基因（淋巴管生成的主要调节因子）来实验性预防淋巴水肿。将TNT应用于先前优化的淋巴水肿小鼠尾巴（第0天），直接在手术部位，将基因货物装载到TNT储存器中：第一组（假手术组）给予pCMV6（仅表达载体骨架），第二组用pCMV6-处理。与假手术组相比，第二组小鼠尾巴体积减小（47.8%），淋巴管造影显示淋巴清除率更高。免疫组织化学显示第二组与第一组相比淋巴管密度更高，RNA测序显示炎症标志物减少。与对照组相比，在损伤当天使用TNT预防性递送至手术部位可减少小鼠尾巴模型中淋巴水肿的表现。
