Abstract
Nuclear proteins, like histone H2A, are promising non-viral carriers for gene delivery since they are biocompatible, biodegradable, bear intrinsic nuclear localization signal, and are easy to modify. The addition of surface-protein-binding ligand to histone H2A may increase its DNA delivery efficiency. Tumor microenvironment (TME) is a promising target for gene therapy since its surface protein repertoire is more stable than that of cancer cells. Cancer-associated fibroblasts (CAFs) are important components of TME, and one of their surface markers is beta-type platelet-derived growth factor receptor (PDGFRβ). In this study, we fused histone H2A with PDGFRβ-binding peptide, YG2, to create a novel non-viral fibroblast-targeting DNA carrier, H2A-YG2. The transfection efficiency of histone complexes with pDNA encoding a bicistronic reporter (enhanced green fluorescent protein, EGFP, and firefly luciferase) in PDGFRβ-positive and PDGFRβ-negative cells was estimated by luciferase assay and flow cytometry. The luciferase activity, percentage of transfected cells, and overall EGFP fluorescence were increased due to histone modification with YG2 only in PDGFRβ-positive cells. We also estimated the internalization efficiency of DNA-carrier complexes using tetramethyl-rhodamine-labeled pDNA. The ligand fusion increased DNA internalization only in the PDGFRβ-positive cells. In conclusion, we demonstrated that the H2A-YG2 carrier targeted gene delivery to PDGFRβ-positive tumor stromal cells.
Highlights
Gene therapy, which is based on the delivery of nucleic acids to target cells in the patient’s body, is a promising approach for cancer treatment
H2A-YG2 was expressed in E. coli and purified using ion-exchange chromatography (IEC), followed by reversed-phase
The target protein-containing fractions from IEC and reversed-phase HPLC (RP-HPLC) are shown in Polymers 2020, 12, x FOR PEER REVIEW
Summary
Gene therapy, which is based on the delivery of nucleic acids to target cells in the patient’s body, is a promising approach for cancer treatment. Viral delivery systems are most commonly used in gene therapy systems. They provide an efficient means of transport of genetic material into the target cells owing to their ability to transfer their genome into the cells of the host. Viral carriers have several disadvantages, including cytotoxicity, potential carcinogenicity, difficulties in production and storage, as well as immunogenicity-related problems associated with the re-administration of such drugs [1,2]. In this regard, non-viral delivery systems, based on cationic polymers, cationic. Nuclear proteins have a number of additional advantages, including biocompatibility, biodegradability, ability to deliver the cargo to the nucleus due to intrinsic nuclear localization signal (NLS), and, ease of desired sequence changes
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
