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Method for targeting Hsp27 to inhibit enterovirus type A71 infection and related applications

A virus infection, EV-A71 technology, applied in the direction of antiviral agents, biochemical equipment and methods, pharmaceutical formulations, etc., can solve the problem of limited, promoting or limiting EV-A71 host factors have not been fully determined, host-virus interaction The underlying mechanism is complex and other issues

Inactive Publication Date: 2020-05-08
香港城市大学深圳研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mechanisms underlying host-virus interactions are complex
[0005] Despite the important role of host cell proteins in the viral life cycle, prior art knowledge of how host proteins participate in and regulate EV-A71 infection is still very limited
Furthermore, despite numerous efforts, the host factors involved in promoting or limiting EV-A71 infection have not been fully identified

Method used

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  • Method for targeting Hsp27 to inhibit enterovirus type A71 infection and related applications
  • Method for targeting Hsp27 to inhibit enterovirus type A71 infection and related applications
  • Method for targeting Hsp27 to inhibit enterovirus type A71 infection and related applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1, EV-A71 infection and 2-DE analysis of uninfected RD cells

[0069] RD cells, which have been widely used to study the mechanism of enterovirus infection, were used for the proteomic analysis in this example. Prior art studies on the kinetics of EV-A71 infection have shown that viral RNA synthesis and protein translation are highly active in phage 6 to 9 hours post-infection (p.i.) when cells are infected at a multiplicity of infection (MOI) of 1 or 10; In addition, the progeny virions were gradually packaged and released during the same period (Lu, J., He, Y.Q., Yi, L.N., Zan, H., Kung, H.F. and He, M.L. (2011) Viral kinetics of enterovirus 71 in human abdominal osarcoma cells . World journal of gastroenterology, 17, 4135-4142). In the present invention, in order to identify host factors that respond to early viral replication, proteins were extracted and p.i. applied to proteomic analysis after 6 hours. At this point, EV-A71 infection is established with h...

Embodiment 2

[0075] Embodiment 2, western blot analysis protein

[0076] Western blot analysis was performed using whole protein extracts of RD cells that were mock or EV-A71-infected at an MOI of 10 at 6 hours post-infection. Three altered chaperones (GRp78, PHB and Hsp27) associated with cellular stress responses were selected for validation. Consistent with the observations in the 2-DE analysis, all three proteins were found to be upregulated ( Figure 1C , Figure 1D ).

Embodiment 3

[0077] Example 3, Hsp27 expression up-regulation in cells infected with EV-A71

[0078] In order to reveal the dynamic changes of Hsp27 during EV-A71 infection, this example analyzed the mRNA and protein levels of Hsp27 during virus infection. Cellular mRNA and protein were extracted from EV-A71 infected cells at different time points. Such as Figure 2A As shown, the mRNA level of Hsp27 was significantly up-regulated at p.i. 6 hours and 9 hours, and the up-regulation was about 100% at 6 hours. Likewise, when the viral protein VP1 was expressed at relatively high levels, Hsp27 protein levels also increased by 40% to 50% at 6, 9 and 12 hours p.i. ( Figure 2B ).

[0079] These results were further confirmed in Hela cells ( Figure 3A ), indicating that EV-A71 infection induces Hsp27 expression.

[0080] Furthermore, new synthesis of viral proteins was first detected at 6 hours p.i. A significant increase in Hsp27 was also found at the same time point ( Figure 2B ).

[0...

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Abstract

The invention provides a method for targeting Hsp27 to inhibit enterovirus type A71 infection and related applications. Specifically, applications of Hsp27, as a target spot, to the developing, screening and / or preparing of drugs for controlling and / or inhibiting viral infection are also provided; and viruses are positive-strand RNA viruses containing IRES sequences, such as EV-A71, HCV or dengueviruses. A method for targeting small heat shock protein Hsp27 to inhibit enterovirus A71 infection is also provided.

Description

technical field [0001] The present invention relates to a new application of small heat shock protein Hsp27, specifically, the present invention relates to a method and related application of targeting Hsp27 to inhibit enterovirus A71 (EV-A71) infection. Background technique [0002] Hand-foot-mouth disease (HFMD) is a global concern and is mainly caused by enterovirus A71 (EV-A71) and coxsackievirus A16 (CVA16). HFMD is usually a self-limiting disease with mild clinical manifestations, but in some cases, it can also lead to serious neurological complications and even death in children, especially those under 5 years of age. Complications include poliomyelitis-like acute flaccid paralysis, aseptic meningitis, brainstem encephalitis, and pulmonary edema. [0003] EV-A71 was first isolated in California in 1969, and was identified as the pathogen causing hand, foot and mouth disease in Japan in 1973. In recent decades, sporadic occurrences or outbreaks of EV-A71 infection ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/352A61P31/14C12Q1/686C12Q1/70
CPCA61K31/352A61K45/00A61P31/14C12Q1/686C12Q1/701C12Q2600/158C12Q2521/107C12Q2561/113Y02A50/30
Inventor 何明亮陈缨但雪莲
Owner 香港城市大学深圳研究院
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