Egorova A A, Shtykalova S V, Maretina M A, Selyutin A V, Shved N Yu, Krylova N V, Ilina A V, Pyankov I A, Freund S A, Selkov S A, Baranov V S, Kiselev A V
Ott Research Institute of Obstetrics, Gynecology, and Reproductology, Saint-Petersburg, 199034 Russia.
St. Petersburg State University, St. Petersburg, 199034 Russia.
Mol Biol (Mosk). 2020 May-Jun;54(3):497-511. doi: 10.31857/S0026898420030064.
Uterine leiomyoma (UL) is the most common benign tumor in women of reproductive age. Gene therapy using suicidal genes appears to be a promising approach for UL treatment. One of key factors for success of gene therapy is the right choice of genetic construct carrier. A promising group of non-viral carriers for cell delivery of expression vectors is cationic Cys-flanked peptides which form tight complexes with DNA due to electrostatic interactions and the presence of interpeptide disulfide bonds. The paper reports a comparative study of the physico-chemical, toxic, and transfectional properties of the DNA-peptide complexes obtained by matrix polymerization or oxidative polycondensation of Cys-flanked peptides using the chain growth terminator 2-amino ethanethiol. We have demonstrated the therapeutic effect of the delivery of the pPTK-1 plasmid carrying the herpes simplex virus type 1 (HSV-1) thymidine kinase gene into PANC-1, and HEK-293T cell culture as well as into primary UL cells. It has been shown that the carriers obtained by oxidative polycondensation transform primary UL cells more efficiently than those produced by matrix polymerization. Treatment with ganciclovir resulted in the death of up to 40% of UL cells transfected with the pPTK-1 plasmid. The perspectives of use of the polyR6 carrier produced by oxidative polycondensation as a tool for the development of modular peptide carriers for the purposes of UL gene therapy were discussed.
子宫肌瘤（UL）是育龄女性最常见的良性肿瘤。利用自杀基因进行基因治疗似乎是一种有前景的子宫肌瘤治疗方法。基因治疗成功的关键因素之一是正确选择基因构建载体。一类有前景的用于表达载体细胞递送的非病毒载体是阳离子半胱氨酸侧翼肽，由于静电相互作用和肽间二硫键的存在，它们能与DNA形成紧密复合物。本文报道了一项比较研究，该研究涉及使用链增长终止剂2-氨基乙硫醇通过半胱氨酸侧翼肽的基质聚合或氧化缩聚获得的DNA-肽复合物的物理化学性质、毒性和转染特性。我们已经证明，将携带单纯疱疹病毒1型（HSV-1）胸苷激酶基因的pPTK-1质粒递送至PANC-1、HEK-293T细胞培养物以及原代子宫肌瘤细胞中具有治疗效果。结果表明，通过氧化缩聚获得的载体比通过基质聚合产生的载体更有效地转化原代子宫肌瘤细胞。用更昔洛韦处理导致高达40%的用pPTK-1质粒转染的子宫肌瘤细胞死亡。讨论了将氧化缩聚产生的polyR6载体用作开发用于子宫肌瘤基因治疗的模块化肽载体工具的前景。
