Identification of human immunodeficiency virus type-1 Gag-TSG101 interaction inhibitors by high-throughput screening
Identification of human immunodeficiency virus type-1 Gag-TSG101 interaction inhibitors by high-throughput screening
复制标题
高通量筛选鉴定人类免疫缺陷病毒1型Gag-TSG101相互作用抑制剂
DOI：
10.1016/j.bbrc.2018.08.079
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发表时间：
2018
影响因子：
3.1
通讯作者：
Yoko Aida
中科院分区：
文献类型：
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作者：
Lowela Siarot;Nopporn Chutiwitoonchai;Hirotaka Sato;Hao Chang;Hironori Sato;Masayuki,Fujino,Tsutomu Murakami,Toshihiro Aono;Eiichi Kodama;Kazumichi Kuroda;Masami Takei;Yoko Aida
病毒蛋白GAG与细胞肿瘤易感基因101(TSG101)的相互作用是HIV-1复制周期中的关键步骤。这种相互作用通过运输所需的细胞内体分选复合体(ESCRT)途径启动病毒组装/萌发，使其成为抗病毒治疗的潜在靶点。在这里，我们开发了一个简单、强大和可靠的基于酶联免疫吸附试验(ELISA)的高通量筛选(HTS)系统，通过靶向Gag-TSG101相互作用来鉴定抑制HIV-1复制的化合物。通过使用所建立的HTS系统对9600化合物文库进行筛选，鉴定出几个抑制Gag-TSG101相互作用的HIT化合物。随后的检测发现了两个热门化合物HSM-9和HSM-10，它们对嗜CD4+T细胞的NL4-3和嗜巨噬细胞的JR-CSF HIV-1毒株具有抗病毒活性。这些结果表明，我们建立的HTS系统是鉴定HIV-1 Gag-TSG101相互作用抑制剂不可或缺的工具。
The interaction between viral protein Gag and cellular protein tumor susceptibility gene 101 (TSG101) is a crucial step in the HIV-1 replication cycle. This interaction initiates the viral assembly/budding via the cellular endosomal sorting complexes required for transport (ESCRT) pathway, making it a potential target for antiviral therapy. Here we developed a simple, robust, and reliable high-throughput screening (HTS) system based on enzyme-linked immunosorbent assay (ELISA) to identify compounds that inhibit HIV-1 replication by targeting Gag-TSG101 interaction. Through screening of the 9600-compound library using the established HTS system, several hit compounds, which inhibited Gag-TSG101 interaction, were identified. Subsequent assays revealed two hit compounds, HSM-9 and HSM-10, which have antiviral activity against CD4+T cell-tropic NL4-3 and macrophage-tropic JR-CSF HIV-1 strains. These results suggest that our established HTS system is an indispensable tool for the identification of HIV-1 Gag-TSG101 interaction inhibitors.