Multifunctional peptide conjugated amphiphilic cationic copolymer for enhancing ECs targeting, penetrating and nuclear accumulation
Xinghong Duo1,2, Lingchuang Bai1, Jun Wang1, Jintang Guo1, Xiangkui Ren1,3,4, Shihai Xia5, Wencheng Zhang6, Abraham Domb7, Yakai Feng1,3,4()
1. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China 2. School of Chemistry and Chemical Engineering, Qinghai University for Nationalities, Xining 810007, China 3. Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin), Tianjin 300350, China 4. Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China 5. Department of Hepatopancreatobiliary and Splenic Medicine, Affiliated Hospital, Logistics University of People’s Armed Police Force, Tianjin 300162, China 6. Department of Physiology and Pathophysiology, Logistics University of People’s Armed Police Force, Tianjin 300162, China 7. Institute of Drug Research (IDR), School of Pharmacy-Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 91905, Israel
Gene therapy has drawn great attention in the treatments of many diseases, especially for cardiovascular diseases. However, the development of gene carriers with low cytotoxicity and multitargeting function is still a challenge. Herein, the multitargeting REDV-G-TAT-G-NLS peptide was conjugated to amphiphilic cationic copolymer poly(ε-caprolactone-co-3(S)-methyl-morpholine-2,5-dione)-g-polyethyleneimine (PCLMD-g-PEI) via a heterobifunctional orthopyridyl disulfide-poly(ethylene glycol)-N-hydroxysuccinimide (OPSS-PEG-NHS) linker to prepare PCLMD-g-PEI-PEG-REDV-G-TAT-G-NLS copolymers with the aim to develop the gene carriers with low cytotoxicity and high transfection efficiency. The multitargeting micelles were prepared from PCLMD-g-PEI-PEG-REDV-G-TAT-G-NLS copolymers by self-assembly method and used to load pEGFP-ZNF580 plasmids (pDNA) to form gene complexes for enhancing the proliferation and migration of endothelial cells (ECs). The loading pDNA capacity was proved by agarose gel electrophoresis assay. These multitargeting gene complexes exhibited low cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The high internalization efficiency of these gene complexes was confirmed by flow cytometry. The results of in vitro transfection demonstrated that these multitargeting gene complexes possessed relatively high transfection efficiency. The rapid migration of ECs transfected by these gene complexes was verified by wound healing assay. Owing to ECs-targeting ability, cell-penetrating ability and nuclear targeting capacity of REDV-G-TAT-G-NLS peptide, the multitargeting polycationic gene carrier with low cytotoxicity and high transfection efficiency has great potential in gene therapy.
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