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Application of YAP/TAZ protein as target molecule in preparation of anti-influenza A virus drugs

A type of influenza A virus and target molecule technology, applied in the direction of antiviral agents, DNA/RNA fragments, viral peptides, etc., can solve problems that have not been related to research, to improve expression, improve host antiviral ability, and inhibit intracellular The effect of the ability to replicate

Pending Publication Date: 2022-04-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the regulation of the host YAP / TAZ protein after infection with influenza A virus and the regulation mechanism have not been reported yet.
Whether YAP / TAZ can be used as a new target for anti-influenza A virus infection and replication has not been studied yet

Method used

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  • Application of YAP/TAZ protein as target molecule in preparation of anti-influenza A virus drugs
  • Application of YAP/TAZ protein as target molecule in preparation of anti-influenza A virus drugs
  • Application of YAP/TAZ protein as target molecule in preparation of anti-influenza A virus drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: Analysis of the impact of influenza A virus infection on YAP and TAZ proteins

[0051] In this embodiment, the changes of the expression levels and phosphorylation levels of YAP and TAZ proteins over time after influenza A virus infection were first detected. Spread a strain of human alveolar epithelial cell A549 into a culture dish containing DMEM medium (containing 10% fetal bovine serum and 1% penicillin / streptomycin), and put it in a carbon dioxide incubator (37°C, 5% CO 2 ) cultured to a cell density of approximately 1×10 6 cell / mL, add influenza A virus PR8 (A / PR8 / 34 / H1N1 strain) (MOI=0.01) and continue to co-culture with the cells for 48 hours (the culture conditions are the same as above), and collect at 6, 12, 24, and 48 hours cells, and cells not infected with the virus were used as controls. Collect the cells after centrifugation, add cell lysate and shake to fully lyse the cells, and collect the supernatant after centrifugation again. On the...

Embodiment 2

[0055] Example 2: Analysis of the regulatory effect of influenza A virus non-structural protein NS1 on YAP and TAZ proteins

[0056] In this example, A549 cells were cultured according to the method in Example 1 to a cell density of about 1×10 6 cell / mL, the cells were transfected with a plasmid capable of overexpressing the viral NS1 protein, and the cells not transfected with the plasmid were used as a control, and the cells were collected by centrifugation after continuing to culture for 48 hours. Add cell lysate to the cells and shake to fully lyse the cells, then centrifuge again to collect the supernatant. According to the method in Example 1, the protein in the cell lysed supernatant sample was transferred to a nitrocellulose membrane, and the phosphorylation level and expression level of YAP protein were detected by antibodies. The result is as image 3 As shown, it can be seen that when the NS1 protein is overexpressed in cells, the dephosphorylation level of YAP pr...

Embodiment 3

[0061] Example 3: Inhibition of YAP and / or TAZ can activate TLR3 signaling pathway

[0062] In this embodiment, the changes of TLR3 signaling pathway in A549 cells suppressed YAP and / or TAZ gene were firstly analyzed when infected with influenza A virus.

[0063] Cultivate A549 cells according to the method in Example 1 until the cell density is about 1×10 6 YAP and TAZ gene-specific siRNA were transfected at cell / mL, and the negative control siRNA was used as a control. Influenza A virus PR8 was added to the above two groups of cells according to the multiplicity of infection (MOI=0.01) and infected for 12 h, and cell samples were collected at 0 h, 6 h, and 12 h after infection. The cells were lysed according to the method described in Example 1, and the changes of each protein in the TLR3 signaling pathway were detected by immunoblotting, and the results were as follows Figure 5 shown. It can be seen that compared with the control group, inhibition of YAP and / or TAZ gene ...

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Abstract

The invention relates to application of effect protein YAP and / or TAZ in a Hippo signal channel as a target spot in inhibition of influenza A virus infection and replication. Specifically, the invention discloses an application of YAP and / or TAZ protein as a target molecule in preparation of an anti-influenza A virus drug. By inhibiting the expression level of the YAP and / or TAZ gene, the anti-virus capability of the cell when the cell is infected with the influenza A virus can be improved; the antiviral ability refers to activation of a TLR3 signal channel, reduction of virus replication and multiplication ability and reduction of the damage degree of the virus to lung tissues of experimental animals.

Description

technical field [0001] The invention relates to the use of the effector protein YAP and / or TAZ in the Hippo signaling pathway as a target to inhibit the infection and replication of influenza A virus. Background technique [0002] Influenza A virus (Influenza A virus) is a single-stranded RNA virus belonging to the Orthomyxoviridae family, and it is the pathogen that has caused the global influenza epidemic in recent decades. Although vaccination has been used to prevent influenza virus infection, due to the complexity of influenza virus typing and the limitation of epidemic prediction ability, outbreaks still occur from time to time, and for pregnant women, the elderly, children and some people with chronic diseases or low immunity can cause severe respiratory illness and even death. According to statistics, millions of people around the world are infected with influenza virus every year and hundreds of thousands of people die from severe respiratory diseases related to in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C07K14/11A61K45/00A61K31/713A61P31/16
Inventor 刁宏燕王凯航张琼章旭君
Owner ZHEJIANG UNIV
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