Design of a PROTAC that antagonizes and destroys the cancer-forming X-protein of the hepatitis B virus
Design of a PROTAC that antagonizes and destroys the cancer-forming X-protein of the hepatitis B virus
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DOI：
10.1016/j.bbrc.2014.10.006
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发表时间：
2014-10-31
影响因子：
3.1
通讯作者：
Krissansen, Geoffrey W.
中科院分区：
文献类型：
--
作者：
Montrose, Kristopher;Krissansen, Geoffrey W.
乙型肝炎病毒（HBV）的x蛋白对病毒感染至关重要，并有助于HBV诱导的肝细胞癌（HCC）的发展，这种疾病每年导致100多万人死亡。在这里，我们描述了一种新的PROTAC（蛋白水解靶向嵌合分子）的设计，能够同时诱导x蛋白的降解，并拮抗其功能。PROTAC是通过将x蛋白的n端寡聚和c端不稳定结构域相互融合，并通过包含聚精氨酸细胞穿透肽（CPP）使其具有细胞渗透性而构建的。预测寡聚化结构域将结合x蛋白，而不稳定结构域将导致x蛋白成为蛋白酶体降解的靶标。将PROTAC添加到HepG2肝癌细胞中，工程化表达全长和c端截断形式的x蛋白，导致两种形式的x蛋白降解。HepG2细胞占据了一种细胞可渗透的独立形式的寡聚化结构域，并作为显性阴性抑制剂，导致x蛋白诱导的细胞凋亡受到抑制。总之，本文描述的PROTAC诱导x蛋白的降解，并拮抗其功能，值得在临床前研究中研究其预防或治疗HBV感染和/或HCC发展的能力。(C) 2014爱思唯尔公司版权所有。
The X-protein of the hepatitis B virus (HBV) is essential for virus infection and contributes to the development of HBV-induced hepatocellular carcinoma (HCC), a disease which causes more than one million deaths each year. Here we describe the design of a novel PROTAC (proteolysis targeting chimeric molecule) capable of simultaneously inducing the degradation of the X-protein, and antagonizing its function. The PROTAC was constructed by fusing the N-terminal oligomerization and C-terminal instability domains of the X-protein to each other, and rendering them cell-permeable by the inclusion of a polyarginine cell-penetrating peptide (CPP). It was predicted that the oligomerization domain would bind the X-protein, and that the instability domain would cause the X-protein to be targeted for proteasomal degradation. Addition of the PROTAC to HepG2 liver cancer cells, engineered to express full-length and C-terminally truncated forms of the X-protein, resulted in the degradation of both forms of the X-protein. A cell-permeable stand-alone form of the oligomerization domain was taken up by HepG2 cells, and acted as a dominant-negative inhibitor, causing inhibition of X-protein-induced apoptosis. In summary, the PROTAC described here induces the degradation of the X-protein, and antagonizes its function, and warrants investigation in a preclinical study for its ability to prevent or treat HBV infection and/or the development of HCC. (C) 2014 Elsevier Inc. All rights reserved.