School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China.
Institute of Dermatology and Venereology, Dermatology Hospital, Southern Medical University, Guangzhou, China.
Hum Gene Ther. 2022 Dec;33(23-24):1269-1278. doi: 10.1089/hum.2022.053.
Gene therapy's entrance into clinical settings has made it an ever more attractive field of study for various diseases. However, relatively little progress has been made in targeting kidney diseases due to poor gene delivery efficiency in renal cells. The development of novel gene therapy vectors for medical intervention to treat kidney diseases is needed. In this study, we designed and produced a pseudotyped lentiviral vector with envelope glycoproteins of Zika virus (ZIKV), and evaluated its potential use in viral vector entry, neutralization assay, and gene delivery especially in the renal context. The lentiviral vector, simplified as ZIKV-E, is pseudotyped with Env/G-TC representing the transmembrane (TM) and cytoplasmic (Y) domains of Env replaced with the TM and CY domains of the glycoprotein (G) of the vesicular stomatitis virus. results show that ZIKV-E induced efficient transduction in tubular epithelial cells in mouse kidneys, demonstrating >100-fold higher expression of exogenous green fluorescent protein gene compared with that achieved by vesicular stomatitis virus G (VSV-G) protein pseudotyped lentiviral vector. The results also showed that the vector ZIKV-E transduced cells in a pH-independent manner and the transduction was inhibited by anti-ZIKV Env domain III antibodies. Results also show that ZIKV-E can be used as a surrogate for studies of ZIKV entry mechanisms and neutralization antibody assay. In all, this study successfully demonstrated a novel pseudotyped lentiviral vector ZIKV-E for inducing high transduction efficiency in renal tubular epithelial cells that could serve as a foundation for gene therapy for the treatment of inherited renal diseases in humans.
基因治疗进入临床领域，使其成为各种疾病更具吸引力的研究领域。然而，由于肾细胞中的基因传递效率较差，针对肾脏疾病的相对进展较少。需要开发用于医学干预的新型基因治疗载体来治疗肾脏疾病。在这项研究中，我们设计并生产了一种带有寨卡病毒（ZIKV）包膜糖蛋白的假型慢病毒载体，并评估了其在病毒载体进入、中和测定和基因传递中的潜在用途，特别是在肾脏环境中。该慢病毒载体简化为 ZIKV-E，假型化为 Env/G-TC，代表包膜蛋白（Env）的跨膜（TM）和细胞质（Y）结构域被水疱性口炎病毒（VSV）糖蛋白（G）的 TM 和 CY 结构域取代。结果表明，ZIKV-E 可诱导小鼠肾脏中的管状上皮细胞有效转导，与 VSV-G 蛋白假型慢病毒载体相比，外源绿色荧光蛋白基因的表达高出 100 倍以上。结果还表明，该载体 ZIKV-E 以 pH 独立的方式转导细胞，并且转导被抗 ZIKV 包膜蛋白结构域 III 抗体抑制。结果还表明，ZIKV-E 可用于研究 ZIKV 进入机制和中和抗体测定的替代物。总之，本研究成功地证明了一种新型的假型慢病毒载体 ZIKV-E，它可以在肾小管上皮细胞中诱导高转导效率，可为人类遗传性肾脏疾病的基因治疗奠定基础。
