ROS-responsive nanoparticle-mediated delivery of CYP2J2 gene for therapeutic angiogenesis in severe hindlimb ischemia

Liang Gui; Youlu Chen; Yongpeng Diao; Zuoguan Chen; Jianwei Duan; Xiaoyu Liang; Huiyang Li; Kaijing Liu; Yuqing Miao; Qing Gao; Zhichao Li; Jing Yang; Yongjun Li

doi:10.1016/j.mtbio.2021.100192

ROS-responsive nanoparticle-mediated delivery of CYP2J2 gene for therapeutic angiogenesis in severe hindlimb ischemia

Mater Today Bio. 2021 Dec 20:13:100192. doi: 10.1016/j.mtbio.2021.100192. eCollection 2022 Jan.

Authors

Liang Gui^{1

2

3}, Youlu Chen⁴, Yongpeng Diao¹, Zuoguan Chen¹, Jianwei Duan⁴, Xiaoyu Liang⁴, Huiyang Li⁴, Kaijing Liu⁴, Yuqing Miao¹, Qing Gao¹, Zhichao Li¹, Jing Yang⁴, Yongjun Li¹

Affiliations

¹ Department of Vascular Surgery, Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, 100730, PR China.
² Graduate School of Peking Union Medical College, Beijing, 100730, PR China.
³ Department of Vascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, PR China.
⁴ Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 300192, PR China.

Abstract

With critical limb ischemia (CLI) being a multi-factorial disease, it is becoming evident that gene therapy with a multiple bio-functional growth factor could achieve better therapeutic outcomes. Cytochrome P450 epoxygenase-2J2 (CYP2J2) and its catalytic products epoxyeicosatrienoic acids (EETs) exhibit pleiotropic biological activities, including pro-angiogenic, anti-inflammatory and cardiovascular protective effects, which are considerably beneficial for reversing ischemia and restoring local blood flow in CLI. Here, we designed a nanoparticle-based pcDNA3.1-CYP2J2 plasmid DNA (pDNA) delivery system (nanoparticle/pDNA complex) composed of a novel three-arm star block copolymer (3S-PLGA-po-PEG), which was achieved by conjugating three-armed PLGA to PEG via the peroxalate ester bond. Considering the multiple bio-functions of CYP2J2-EETs and the sensitivity of the peroxalate ester bond to H₂O₂, this nanoparticle-based gene delivery system is expected to exhibit excellent pro-angiogenic effects while improving the high oxidative stress and inflammatory micro-environment in ischemic hindlimb. Our study reports the first application of CYP2J2 in the field of therapeutic angiogenesis for CLI treatment and our findings demonstrated good biocompatibility, stability and sustained release properties of the CYP2J2 nano-delivery system. In addition, this nanoparticle-based gene delivery system showed high transfection efficiency and efficient VEGF expression in vitro and in vivo. Intramuscular injection of nanoparticle/pDNA complexes into mice with hindlimb ischemia resulted in significant rapid blood flow recovery and improved muscle repair compared to mice treated with naked pDNA. In summary, 3S-PLGA-po-PEG/CYP2J2-pDNA complexes have tremendous potential and provide a practical strategy for the treatment of limb ischemia. Moreover, 3S-PLGA-po-PEG nanoparticles might be useful as a potential non-viral carrier for other gene delivery applications.

Keywords: CLI, critical limb ischemia; CYP2J2; CYP2J2, cytochrome P450 epoxygenase 2J2; EETs, epoxyeicosatrienoic acids; Gene therapy; Hindlimb ischemia; Nanocarriers; PAD, peripheral artery disease; ROS, reactive oxygen species; Therapeutic angiogenesis.