Polycondensed Peptide Carriers Modified with Cyclic RGD Ligand for Targeted Suicide Gene Delivery to Uterine Fibroid Cells.

Anna Egorova,Anton Kiselev,Natalia Shved,Sergey Selkov,Sofia Shtykalova,Vladislav Baranov,Alexander Selutin,Dmitriy Deviatkin,Marianna Maretina

doi:10.3390/ijms23031164

Abstract

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.

Highlights

IntroductionThere are three major types of uterine fibroids (UFs)—submucosal, subserosal, and intramural fibroids [4]
Most studies on suicide gene therapy use the delivery of the herpes simplex virus thymidine kinase (HSV-TK) gene followed by treatment with prodrugs, such as acyclovir or ganciclovir (GCV) [8]
3-nitro-2-pyridinesulfenyl group (Npys), which acts as an activating group for disulfide bond formation, as it is displaced by the free thiol during oxidative polycondensation of R6 monomers [38]

Summary

Introduction

There are three major types of UF—submucosal, subserosal, and intramural fibroids [4] The latter grow within the muscular uterine wall and imply certain difficulties and increased traumatism of the healthy myometrium during myomectomy, which makes this approach undesirable for treatment of intramural fibroids in comparison with hormonal therapy [5]. A significant advantage of this approach is the so-called “bystander effect”, the phenomenon based on the fact that phosphorylated GCV enters neighboring non-transfected cells through intercellular contacts widely spread in UF cells compared to normal myometrium; this increases the efficiency of therapy without negative effects on healthy tissues [9,10]. The proposed gene therapy approach may be beneficial for UF treatment, especially for fibroids with an intramural location

Methods

Results

Conclusion