A Feasible Alternative Strategy Targeting Furin Disrupts SARS-CoV-2 Infection Cycle.
A Feasible Alternative Strategy Targeting Furin Disrupts SARS-CoV-2 Infection Cycle.
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一种可行的替代策略靶向弗林蛋白酶破坏SARS-CoV-2感染周期。
DOI：
10.1128/spectrum.02364-21
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发表时间：
2022-02-23
影响因子：
3.7
通讯作者：
Tushir-Singh J
中科院分区：
文献类型：
--
作者：
Mondal T;Shivange G;Habieb A;Tushir-Singh J
COVID-19引起的冠状病毒（SARS-CoV-2）仍然是全球公共卫生威胁。SARS-CoV-2通过其刺突糖蛋白结合血管紧张素转换酶2（ACE 2）进入人肺细胞。值得注意的是，病毒产生细胞中宿主细胞蛋白酶弗林蛋白酶对刺突的切割对于随后刺突驱动进入肺细胞是至关重要的。因此，阻断病毒产生细胞中的刺突裂解和活化的有效靶向疗法可以提供打破病毒传播周期和克服疾病病理的替代策略。在这里，我们设计并描述了一种基于抗体的靶向策略，该策略直接与弗林蛋白酶介导的病毒产生细胞中刺突的蛋白水解激活竞争。所描述的方法涉及在SARS-CoV-2刺突靶向抗体的IgG 1 Fc延伸的柔性接头结构域中工程化竞争性弗林蛋白酶底物残基。考虑到刺突弗林蛋白酶切割和SARS-CoV-2外出的位点仍然不确定，这里追求的实验策略揭示了刺突蛋白的蛋白水解加工的新机制见解，这表明加工不发生在组成性分泌途径中。此外，我们的研究结果表明，阻断弗林蛋白酶介导的切割的刺突蛋白膜融合激活和病毒宿主细胞进入功能。这些发现为SARS-CoV-2和未来冠状病毒科家族成员的靶向适用性提供了另一种见解，利用宿主蛋白酶系统获得细胞进入和随后的感染链。重要性自2019年12月出现以来，COVID-19一直是全球经济及健康威胁。尽管基于RNA和DNA载体的疫苗诱导的抗体应答和免疫记忆已被证明对住院治疗和死亡率非常有效，但它们对不断进化的SARS-CoV-2变体的长期有效性仍然未知。由于宿主细胞富集的弗林蛋白酶介导的SARS-CoV-2刺突蛋白的切割对于病毒进入和感染循环链至关重要，因此本文描述的抗体Fc缀合的弗林蛋白酶竞争肽的解决方案是重要的。在刺突突变漂移不干扰Fc缀合抗体的表位的情况下，所提出的弗林蛋白酶竞争策略赋予广谱靶向设计以阻止有效传播的SARS-CoV-2病毒颗粒的产生。此外，所提出的方法是针对其他潜在致命病毒的即插即用，这些病毒利用分泌途径独立的宿主蛋白酶机制来获得细胞进入并随后传输到宿主细胞。
The COVID-19 causing coronavirus (SARS-CoV-2) remains a public health threat worldwide. SARS-CoV-2 enters human lung cells via its spike glycoprotein binding to angiotensin-converting enzyme 2 (ACE2). Notably, the cleavage of the spike by the host cell protease furin in virus-producing cells is critical for subsequent spike-driven entry into lung cells. Thus, effective targeted therapies blocking the spike cleavage and activation in viral producing cells may provide an alternate strategy to break the viral transmission cycle and to overcome disease pathology. Here we engineered and described an antibody-based targeted strategy, which directly competes with the furin mediated proteolytic activation of the spike in virus-producing cells. The described approach involves engineering competitive furin substrate residues in the IgG1 Fc-extended flexible linker domain of SARS-CoV-2 spike targeting antibodies. Considering the site of spike furin cleavage and SARS-CoV-2 egress remains uncertain, the experimental strategy pursued here revealed novel mechanistic insights into proteolytic processing of the spike protein, which suggest that processing does not occur in the constitutive secretory pathway. Furthermore, our results show blockade of furin-mediated cleavage of the spike protein for membrane fusion activation and virus host-cell entry function. These findings provide an alternate insight of targeting applicability to SARS-CoV-2 and the future coronaviridae family members, exploiting the host protease system to gain cellular entry and subsequent chain of infections. IMPORTANCE Since its emergence in December 2019, COVID-19 has remained a global economic and health threat. Although RNA and DNA vector-based vaccines induced antibody response and immunological memory have proven highly effective against hospitalization and mortality, their long-term efficacy remains unknown against continuously evolving SARS-CoV-2 variants. As host cell-enriched furin-mediated cleavage of SARS-CoV-2 spike protein is critical for viral entry and chain of the infection cycle, the solution described here of an antibody Fc-conjugated furin competing peptide is significant. In a scenario where spike mutational drifts do not interfere with the Fc-conjugated antibody's epitope, the proposed furin competing strategy confers a broad-spectrum targeting design to impede the production of efficiently transmissible SARS-CoV-2 viral particles. In addition, the proposed approach is plug-and-play against other potentially deadly viruses that exploit secretory pathway independent host protease machinery to gain cellular entry and subsequent transmissions to host cells.
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影响因子：
5.4
作者：
Kleine-Weber H;Elzayat MT;Wang L;Graham BS;Müller MA;Drosten C;Pöhlmann S;Hoffmann M
通讯作者：
Hoffmann M
影响因子：
56.9
作者：
Cai, Yongfei;Zhang, Jun;Chen, Bing
通讯作者：
Chen, Bing
影响因子：
14.9
作者：
Cuellar TL;Barnes D;Nelson C;Tanguay J;Yu SF;Wen X;Scales SJ;Gesch J;Davis D;van Brabant Smith A;Leake D;Vandlen R;Siebel CW
通讯作者：
Siebel CW
DOI：
10.1073/pnas.2016093117
发表时间：
2020-11-10
影响因子：
11.1
作者：
Glasgow A;Glasgow J;Limonta D;Solomon P;Lui I;Zhang Y;Nix MA;Rettko NJ;Zha S;Yamin R;Kao K;Rosenberg OS;Ravetch JV;Wiita AP;Leung KK;Lim SA;Zhou XX;Hobman TC;Kortemme T;Wells JA
通讯作者：
Wells JA
β-核心就是使用溶酶体进行出口，而不是生物合成分泌途径。
影响因子：
64.5
作者：
Ghosh S;Dellibovi-Ragheb TA;Kerviel A;Pak E;Qiu Q;Fisher M;Takvorian PM;Bleck C;Hsu VW;Fehr AR;Perlman S;Achar SR;Straus MR;Whittaker GR;de Haan CAM;Kehrl J;Altan-Bonnet G;Altan-Bonnet N
通讯作者：
Altan-Bonnet N