Department of Genomic Medicine, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint Petersburg, Russia.
Department of Immunology and Intercellular Interactions, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint Petersburg, Russia.
Int J Mol Sci. 2022 Jan 21;23(3):1164. doi: 10.3390/ijms23031164.
Suicide gene therapy was suggested as a possible strategy for the treatment of uterine fibroids (UFs), which are the most common benign tumors inwomen of reproductive age. For successful suicide gene therapy, DNAtherapeutics should be specifically delivered to UF cells. Peptide carriers are promising non-viral gene delivery systems that can be easily modified with ligands and other biomolecules to overcome DNA transfer barriers. Here we designed polycondensed peptide carriers modified with a cyclic RGD moiety for targeted DNA delivery to UF cells. Molecular weights of the resultant polymers were determined, and inclusion of the ligand was confirmed by MALDI-TOF. The physicochemical properties of the polyplexes, as well as cellular DNA transport, toxicity, and transfection efficiency were studied, and the specificity of αvβ3 integrin-expressing cell transfection was proved. The modification with the ligand resulted in a three-fold increase of transfection efficiency. Modeling of the suicide gene therapy by transferring the HSV-TK suicide gene to primary cells obtained from myomatous nodes of uterine leiomyoma patients was carried out. We observed up to a 2.3-fold decrease in proliferative activity after ganciclovir treatment of the transfected cells. Pro- and anti-apoptotic gene expression analysis confirmed our findings that the developed polyplexes stimulate UF cell death in a suicide-specific manner.
自杀基因治疗被认为是治疗子宫肌瘤(UFs)的一种可能策略,UFs 是育龄妇女中最常见的良性肿瘤。为了成功进行自杀基因治疗,DNA 治疗剂应专门递送到 UF 细胞。肽载体是很有前途的非病毒基因传递系统,可以很容易地用配体和其他生物分子进行修饰,以克服 DNA 传递障碍。在这里,我们设计了带有环状 RGD 部分的聚缩肽载体,用于将 DNA 靶向递送到 UF 细胞。测定了所得聚合物的分子量,并通过 MALDI-TOF 证实了配体的包含。研究了聚集体的理化性质以及细胞内 DNA 转运、毒性和转染效率,并证明了表达 αvβ3 整合素的细胞转染的特异性。配体的修饰导致转染效率提高了三倍。通过将 HSV-TK 自杀基因转移到从子宫平滑肌瘤患者肌瘤结节获得的原代细胞中,对自杀基因治疗进行了建模。我们观察到转染细胞用更昔洛韦处理后增殖活性下降了 2.3 倍。促凋亡和抗凋亡基因表达分析证实了我们的发现,即开发的聚集体以自杀特异性方式刺激 UF 细胞死亡。