Cys-Flanked Cationic Peptides For Cell Delivery of the Herpes Simplex Virus Thymidine Kinase Gene for Suicide Gene Therapy of Uterine Leiomyoma

Egorova, Anna; Shtykalova, Sofia; Maretina, Marianna; Selyutin, A. V.; Shved, Natallia; Krylova, Nadezhda; Ilina, Arina; Pyankov, Ivan; Freund, Svetlana; Selkov, S. A.; Baranov, Vladislav S; Kiselev, Anton

doi:10.1134/s0026893320030061

Cited by 5 publications

(9 citation statements)

References 51 publications

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“…Primary leiomyoma cells were obtained after myomectomy in the D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology (Saint-Petersburg, Russia) as previously reported [ 43 ]. Briefly, dissected collagenase IV treated fibroids were resuspended in AmnioMax Basal Medium with 10% AmnioMax Supplement serum (Thermo Fisher Scientific, Carlsbad, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

“…The expression of αvβ3 integrins in leiomyoma cells was determined by flow cytometry after cells detaching from flasks with 5 mM EDTA in 1× phosphate-buffered saline (PBS) (Rosmedbio, Saint-Petersburg, Russia) and staining with FITC mouse anti-human CD51/CD61 antibodies (BD Pharmingen, San Jose, CA, USA) for 20 min at room temperature [ 43 ]. Flow cytometry was conducted using BD FACS-Canto II cytofluorimeter (Becton-Dickinson Biosciences, Franklin Lakes, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

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Development of iRGD-Modified Peptide Carriers for Suicide Gene Therapy of Uterine Leiomyoma

et al. 2021

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Uterine leiomyoma (UL) is one of the most common benign tumors in women that often leads to many reproductive complications. Suicide genetherapy was suggested as a promising approach for UL treatment. In the present study, we describe iRGD ligand-conjugated cysteine-rich peptide carrier RGD1-R6 for targeted DNA delivery to αvβ3 integrin-expressing primary UL cells. The physico-chemical properties, cytotoxicity, transfection efficiency and specificity of DNA/RGD1-R6 polyplexes were investigated. TheHSV-1thymidine kinase encoding plasmid delivery to PANC-1pancreatic carcinoma cells and primary UL cells resulted in significant suicide gene therapy effects. Subsequent ganciclovir treatment decreased cells proliferative activity, induced of apoptosis and promoted cells death.The obtained results allow us to concludethatthe developed RGD1-R6 carrier can be considered a promising candidate for suicide gene therapy of uterine leiomyoma.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Development of iRGD-Modified Peptide Carriers for Suicide Gene Therapy of Uterine Leiomyoma

et al. 2021

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“…The efficacy of suicide gene therapy after delivery of pPTK-1 plasmid and pCMV-lacZ (as a control) was accessed in PANC-1 cells in the presence of ganciclovir as previously reported [ 50 ]. Briefly, PANC-1 cells were placed into 96-well cultural plates at a rate of 1.5 × 10 4 cells per well in the standard culture medium, grown for 24 h, and washed with the FBS-free medium.…”

Section: Methodsmentioning

confidence: 99%

Characterization of iRGD-Ligand Modified Arginine-Histidine-Rich Peptides for Nucleic Acid Therapeutics Delivery to αvβ3 Integrin-Expressing Cancer Cells

et al. 2020

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Efficient and specific delivery of nucleic acid (NA) therapeutics to tumor cells is extremely important for cancer gene therapy. Various therapeutic strategies include delivery of DNA-therapeutics such as immunostimulatory or suicide genes and delivery of siRNA-therapeutics able to silence expression of cancer-related genes. Peptides are a promising class of non-viral vehicles which are biodegradable and can efficiently condense, protect and specifically deliver NA to the cells. Here we designed arginine-histidine-rich peptide carriers consisting of an iRGD ligand to target αvβ3 integrins and studied them as vehicles for DNA and siRNA delivery to cancer cells. Combination of iRGD-modified and unmodified arginine–histidine-rich peptides during NA complexation resulted in carriers with different ligand contents. The NA-binding and protecting properties in vitro transfection efficiency and cytotoxicity of the DNA- and siRNA-polyplexes were studied and the most efficient carrier RGD1 was determined. The ability of the peptides to mediate specific intracellular uptake was confirmed inhuman cervical carcinoma (HeLa), human kidney (293T) and human pancreatic (PANC-1) cell lines with different αvβ3 integrins surface expression. By means of RGD1 carrier, efficient delivery of the herpes simplex virus (HSV-1) thymidine kinase gene to PANC-1 cells was demonstrated. Subsequent ganciclovir treatment led to a reduction of PANC-1 cells’ viability by up to 54%. Efficient RNAi-mediated down-regulation of GFP and VEGFA gene expression was achieved in MDA-MB-231-GFP+ breast cancer and EA.hy926 endothelial cells, respectively, by means of RGD1/siRNA polyplexes. Here we demonstrated that the peptide carrier RGD1 can be considered as promising candidate for development of NA therapeutics delivery systems useful in cancer gene therapy.

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“…Primary UL cells were isolated from UL nodes after myomectomy at the D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology (Saint-Petersburg, Russia), as previously reported [22,23]. The UL cells were cultured in AmnioMAX C-100 Complete Medium (Thermo Fisher Scientific, Carlsbad, CA, USA) and transfection experiments were held in DMEM-F12 (Thermo Fisher Scientific) supplemented with 10% FBS (Thermo Fisher Scientific) and 0.3% gentamicin.…”

Section: Cell Linesmentioning

confidence: 99%

“…The DNAse I protection assay requires 10 µL of the peptide/DNA/MNPs polyplexes prepared with different amounts of MNPs and incubated with 0.5 units of DNase I (Ambion, Austin, TX, USA) for 30 min at 37 • C followed by 2 min of DNAse I activation, as described previously [22]. DNA was released from polyplexes with overnight 0.1% trypsin incubation at 37 • C. The DNA integrity was analyzed by 1% agarose gel electrophoresis.…”

Section: Dna Binding and Dnase I Protection Assaymentioning

confidence: 99%

Magnetic Nanoparticles as a Component of Peptide-Based DNA Delivery System for Suicide Gene Therapy of Uterine Leiomyoma

et al. 2022

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Suicidegene therapy is considered a promising approach for the treatment of uterine leiomyoma (UL), a benign tumor in women characterized by precise localization. In this study, we investigate the efficiency of αvβ3 integrin-targeted arginine-rich peptide carrier R6p-cRGD electrostatically bound to magnetic nanoparticles (MNPs) for targeted DNA delivery into the UL cells. The physico–chemical and cytotoxic properties, transfection efficiency, and specificity of R6p-cRGD/DNA/MNPs polyplexes were evaluated. The addition of MNPs resulted in a decrease in the time needed for successful transfection with simultaneous increase in efficiency. We revealed a therapeutic effect on primary UL cells after delivery of plasmid encoding the herpes simplex virus type 1 (HSV-1) thymidine kinase gene. Treatment with ganciclovir resulted in 20% efficiency of suicide gene therapy in UL cells transfected with the pPTK-1 plasmid. Based on these results, we conclude that the use of cationic peptide carriers with MNPs can be promising for the development of modular non-viral carriers for suicide gene delivery to UL cells.

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Cys-Flanked Cationic Peptides For Cell Delivery of the Herpes Simplex Virus Thymidine Kinase Gene for Suicide Gene Therapy of Uterine Leiomyoma

Cited by 5 publications

References 51 publications

Development of iRGD-Modified Peptide Carriers for Suicide Gene Therapy of Uterine Leiomyoma

Development of iRGD-Modified Peptide Carriers for Suicide Gene Therapy of Uterine Leiomyoma

Characterization of iRGD-Ligand Modified Arginine-Histidine-Rich Peptides for Nucleic Acid Therapeutics Delivery to αvβ3 Integrin-Expressing Cancer Cells

Magnetic Nanoparticles as a Component of Peptide-Based DNA Delivery System for Suicide Gene Therapy of Uterine Leiomyoma

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