Efremov A M, Dukhovlinov I V, Dizhe E B, Burov S V, Leko M V, Akif'ev B N, Mogilenko D A, Ivanov I A, Perevozchikov A P, Orlov S V
Tsitologiia. 2010;52(5):371-9.
The rate and character of skin tissue regeneration after wounds, burns and other traumas depend on the cell proliferation within damaged area. Acceleration of healing by stimulation of cell proliferation and extracellular matrix synthesis is one of the most important tasks of modern medicine. There are gene therapy approaches to wound treatment consisting in the transfer of genes encoding mitogenic growth factors to wound area. The most important step in the development of gene therapy approaches is the design of gene delivery tools. In spite of high efficacy of viral vectors, the non-viral means have some preferences (low toxicity, low immunogenity, safety and the absence of backside effects). Among non-viral gene delivery tools, molecular conjugates are the most popular because of their efficacy, simplicity, and the capacity to the targeted gene transfer. In the present work we have developed two molecular conjugates--NLS-TSF7 and NLS-TSF12 consisting of the modified signal of nuclear localization of T-antigen of SV40 virus (cationic part) and the peptide ligands of mammalian transferrin receptor (ligand part). These conjugates bind to plasmid DNA with formation of polyelectrolytic complexes and are capable to deliver plasmid DNA into cells expressing transferrin receptors by receptor-mediated endocytosis. Transfer of the expression vector of luciferase gene in the complex with molecular conjugate NLS-TSF7 to murine surface tissues led to about 100 fold increasing of luciferase activity in comparison with the transfer of free expression vector. Treatment of slash wounds in mice with the complexes of expression vector of synthetic human gene encoding insulin-like growth factor 1 with molecular conjugates NLS-TSF7 led to acceleration of healing in comparison with mice treated with free expression vector. The results obtained confirm the high efficiency of the developed regenerative gene therapy approach for the treatment of damaged skin tissues in mammals.
伤口、烧伤及其他创伤后皮肤组织再生的速度和特性取决于受损区域内的细胞增殖。通过刺激细胞增殖和细胞外基质合成来加速愈合是现代医学最重要的任务之一。存在一些基因治疗方法用于伤口治疗，包括将编码促有丝分裂生长因子的基因转移至伤口区域。基因治疗方法开发中最重要的一步是基因递送工具的设计。尽管病毒载体具有高效性，但非病毒方法有一些优势（低毒性、低免疫原性、安全性及无副作用）。在非病毒基因递送工具中，分子缀合物因其有效性、简便性及靶向基因转移能力而最为常用。在本研究中，我们开发了两种分子缀合物——NLS - TSF7和NLS - TSF12，它们由SV40病毒T抗原的核定位修饰信号（阳离子部分）和哺乳动物转铁蛋白受体的肽配体（配体部分）组成。这些缀合物与质粒DNA结合形成聚电解质复合物，并能够通过受体介导的内吞作用将质粒DNA递送至表达转铁蛋白受体的细胞中。与游离表达载体的转移相比，将荧光素酶基因表达载体与分子缀合物NLS - TSF7形成的复合物转移至小鼠体表组织后，荧光素酶活性提高了约100倍。用编码胰岛素样生长因子1的合成人类基因表达载体与分子缀合物NLS - TSF7形成的复合物治疗小鼠的切割伤口，与用游离表达载体治疗的小鼠相比，伤口愈合加速。所得结果证实了所开发的再生基因治疗方法在治疗哺乳动物受损皮肤组织方面的高效性。
