Guanine nucleotide exchange factor DOCK11-binding peptide fused with a single chain antibody inhibits hepatitis B virus infection and replication.
Guanine nucleotide exchange factor DOCK11-binding peptide fused with a single chain antibody inhibits hepatitis B virus infection and replication.
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与单链抗体融合的鸟嘌呤核苷酸交换因子DOCK 11结合肽抑制B型肝炎病毒感染和复制。
DOI：
10.1016/j.jbc.2022.102097
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发表时间：
2022-07
影响因子：
4.8
通讯作者：
Yanagawa, Hiroshi
中科院分区：
文献类型：
--
作者：
Ide, Mayuko;Tabata, Noriko;Yonemura, Yuko;Shirasaki, Takayoshi;Murai, Kazuhisa;Wang, Ying;Ishida, Atsuya;Okada, Hikari;Honda, Masao;Kaneko, Shuichi;Doi, Nobuhide;Ito, Satoru;Yanagawa, Hiroshi
B型肝炎病毒（HBV）感染是一个主要的全球性健康问题，目前尚无确定的治疗方法。胞质分裂贡献因子11（DOCK 11）是Cdc 42的鸟嘌呤核苷酸交换因子（GEF），对HBV的维持至关重要。然而，针对DOCK 11的潜在治疗策略尚未被探索。我们以前已经开发了一种体外病毒方法，作为蛋白质组学和进化蛋白质工程分析的更有效的工具。在这项研究中，使用体外病毒方法，我们筛选和鉴定了一种新的抗去唾液酸糖蛋白受体（ASGR）抗体，ASGR 3 - 10 M，和DOCK 11结合肽，DCS 8 - 42 A，用于HBV感染的潜在用途。我们进一步构建了由ASGR 3 - 10 M、DCS 8 - 42 A、融合肽和核定位信号组成的融合蛋白（10 M-D42 AN），以将肽递送到肝细胞内。我们使用免疫荧光染色显示，10 M-D42 AN被内吞到早期内体中并释放到细胞质和细胞核中。由于DCS 8 - 42 A与活化的cdc 42相关激酶1（Ack 1）共享同源性，其促进HBV感染所需的EGFR内吞作用，我们还发现10 M-D42 AN抑制EGFR和Ack 1的内吞作用。此外，我们发现10 M-D42 AN抑制了DOCK 11在宿主DNA修复系统中的功能，该系统是共价闭合环状DNA合成所需的，并抑制了小鼠中的HBV增殖。总之，本研究使用抗ASGR抗体、融合肽和DOCK 11结合肽实现了一种新型肝细胞特异性药物递送系统，为HBV提供了一种新的治疗方法。
Hepatitis B virus (HBV) infection is a major global health problem with no established cure. Dedicator of cytokinesis 11 (DOCK11), known as a guanine nucleotide exchange factor (GEF) for Cdc42, is reported to be essential for the maintenance of HBV. However, potential therapeutic strategies targeting DOCK11 have not yet been explored. We have previously developed an in vitro virus method as a more efficient tool for the analysis of proteomics and evolutionary protein engineering. In this study, using the in vitro virus method, we screened and identified a novel antiasialoglycoprotein receptor (ASGR) antibody, ASGR3-10M, and a DOCK11-binding peptide, DCS8-42A, for potential use in HBV infection. We further constructed a fusion protein (10M-D42AN) consisting of ASGR3-10M, DCS8-42A, a fusogenic peptide, and a nuclear localization signal to deliver the peptide inside hepatocytes. We show using immunofluorescence staining that 10M-D42AN was endocytosed into early endosomes and released into the cytoplasm and nucleus. Since DCS8-42A shares homology with activated cdc42-associated kinase 1 (Ack1), which promotes EGFR endocytosis required for HBV infection, we also found that 10M-D42AN inhibited endocytosis of EGFR and Ack1. Furthermore, we show 10M-D42AN suppressed the function of DOCK11 in the host DNA repair system required for covalently closed circular DNA synthesis and suppressed HBV proliferation in mice. In conclusion, this study realizes a novel hepatocyte-specific drug delivery system using an anti-ASGR antibody, a fusogenic peptide, and DOCK11-binding peptide to provide a novel treatment for HBV.
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影响因子：
168.9
作者：
Lozano, Rafael;Naghavi, Mohsen;Murray, Christopher J. L.
通讯作者：
Murray, Christopher J. L.
影响因子：
4.1
作者：
Doi, Nobuhide;Takashima, Hideaki;Yanagawa, Hiroshi
通讯作者：
Yanagawa, Hiroshi
影响因子：
5.4
作者：
Macovei, Alina;Petrareanu, Catalina;Branza-Nichita, Norica
通讯作者：
Branza-Nichita, Norica
影响因子：
64.8
作者：
MANSER, E;LEUNG, T;LIM, L
通讯作者：
LIM, L
影响因子：
7.6
作者：
Guo JT;Guo H
通讯作者：
Guo H