Anionic coated peptide-based carriers as a delivery system for gene therapy of uterine leiomyoma

Uterine leiomyoma (UL) is the most common benign tumor of the female reproductive tract. Despite being benign, this tumor can seriously affect reproductive function and lead to infertility. Localization makes the disease a perfect target for gene therapy. However, excessive extracellular matrix (ECM) remains challenging for gene delivery by nanoparticles. Anionic coating can improve stability of polyplexes and contribute to successful ECM crossing. We developed avb3-integrin-targeted peptide-based carrier and anionic peptide coating for DNA delivery into primary UL cells and UL nodes.

Arginine-histidine-rich peptide carriers modified with cycloRGD-ligand were synthesized. Physicochemical properties of anionic coated DNA-polyplexes were tested. Polyplexes stability was evaluated in transfection experiments in the presence of serum on PANC-1 cells. Suicidal gene therapy with HSV1-TK delivery and subsequent ganciclovir treatment was held for primary UL cells obtained after myomectomy. Efficiency of DNA delivery into UL nodes was demonstrated by GFP detection by fluorescent microscopy.

Anionic peptide coating increased stability of polyplexes and allowed successful transfection in the presence of serum. TrypanBlue staining assay showed decline of proliferative activity among primary UL cells transfected with HSV1-TK carrying complexes in comparison with control lacZ transfected cells. Fluorescence signal was detected on sections of nodes injected with GFP-polyplexes.

Here we showed that avb3-integrin-targeted peptide-based carriers with anionic peptide coating demonstrate high specificity and transfection efficacy of UL cells and successful gene delivery in UL nodes. Further development of anionic coated peptide carriers is promising for UL gene therapy.

Study was supported by RSF grant 21-15-00111.