Application of TDP-43 inhibitor in preparation of medicine for preventing and / or treating asthma or medicine for treating influenza A infection

Abstract

The invention provides an application of a TDP-43 inhibitor in preparation of a medicine for preventing and / or treating asthma or a medicine for treating influenza A infection. The effect of the TDP-43 in prevention or improvement of asthma and influenza A virus infection is found for the first time, and experiments verify that inhibition of TDP-43 expression not only can significantly relieve symptoms of asthma, but also can resist influenza A virus infection during asthma attack, has a very good asthma disease treatment potential, and can be used for preparing medicines for preventing or improving asthma and influenza A virus infection. And a new effective treatment scheme is expected to be provided for asthma and acute attack of asthma complicated by asthma and respiratory virus infection.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and more specifically, to the use of TDP-43 inhibitors in the preparation of medicaments for the prevention and / or treatment of asthma or influenza A infection. Background Technology

[0002] Asthma is a heterogeneous disease characterized by chronic airway inflammation and airway hyperresponsiveness. It is one of the most common chronic diseases in the world, with varying incidence rates ranging from 1% to 18% across different countries. In my country, the prevalence of asthma among adults is approximately 1.24%, and it is showing an increasing trend year by year. Viral infections during asthma attacks often exacerbate the symptoms of asthma patients.

[0003] Currently, there is no drug that can completely cure asthma. First-line clinical treatments for asthma primarily consist of bronchodilates and medications that reduce airway inflammation, such as corticosteroids (e.g., budesonide), β2-receptor agonists (e.g., salbutamol), and leukotriene modifiers (e.g., montelukast). These drugs have strong effects in inhibiting airway inflammation and relaxing airway smooth muscle; however, long-term use can lead to drug resistance and serious adverse reactions, including immunosuppression, metabolic disorders, and osteoporosis.

[0004] In recent years, the development of targeted therapies has received increasing attention in order to improve disease treatment outcomes and reduce side effects. However, current targeted drugs for asthma mostly focus on targets downstream type II inflammation (such as IL-4 / IL-4Rα, IL-5 / IL-5R, IL-13, IgE, etc.), failing to address the challenge of asthma relapse. Furthermore, there is still a lack of specific targeted therapies for acute asthma exacerbations concurrent with respiratory viral infections.

[0005] TDP-43 (TAR DNA-binding protein 43) is a nucleoprotein that plays a crucial role in neurodegenerative diseases. Encoded by the TARDBP gene, its abnormal aggregation in the cytoplasm is a hallmark pathological feature of diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). However, a review of domestic and international literature and materials revealed no reports on the application of TDP-43 in the treatment of chronic lung diseases (such as asthma). Summary of the Invention

[0006] The purpose of this invention is to provide the application of TDP-43 inhibitors in the preparation of drugs for the prevention and / or treatment of asthma or influenza A infection. This invention, through experimental verification, demonstrates that inhibiting TDP-43 expression in the airway epithelium not only significantly alleviates asthma symptoms but also resists influenza A virus infection during asthma attacks, exhibiting excellent therapeutic potential for asthma.

[0007] Based on the above, the present invention first provides the use of TDP-43 inhibitors in the preparation of medicaments for the prevention and / or treatment of asthma.

[0008] Preferably, the asthma includes asthma exacerbations caused by respiratory viral infections.

[0009] Preferably, the TDP-43 inhibitor is a substance that knocks out or knocks down the coding gene for TDP-43, or reduces the expression level or activity of TDP-43 protein, or weakens the effect of TDP-43.

[0010] Preferably, the substance comprises any one of the following: protein, nucleic acid molecule, small molecule compound, or gene intervention tool capable of knocking out or knocking down TDP-43 expression.

[0011] In another aspect, the present invention provides the use of TDP-43 inhibitors in the preparation of medicaments for the prevention and / or treatment of influenza A infection.

[0012] In another aspect, the present invention provides an antisense oligonucleotide targeting the encoding gene of TDP-43, the sequence of which is shown in SEQ ID NO:3 or SEQ ID NO:4.

[0013] Preferably, the antisense oligonucleotide is delivered via a lipid nanoparticle carrier.

[0014] In another aspect, the present invention provides a CRISPR / Cas gene editing system for targeting and knocking out the coding gene of TDP-43, the gene editing system comprising gRNA specifically targeting the TDP-43 gene in mouse airway epithelium, the sequence of which includes SEQ ID NO: 1 and SEQ ID NO: 2.

[0015] In another aspect, the present invention also provides the use of the aforementioned antisense oligonucleotides, or the aforementioned gene editing system, in the preparation of drugs for the prevention and / or treatment of asthma or drugs for the treatment of influenza A infection.

[0016] Preferably, the dosage form of the drug includes nebulizers and drops.

[0017] Compared with the prior art, the beneficial effects of the present invention include at least the following:

[0018] This invention is the first to propose that TDP-43, which is highly expressed in the airway epithelium induced by allergens, is a key upstream therapeutic target for triggering airway hyperresponsiveness and asthma pathogenesis. Through experimental verification, inhibiting the expression of TDP-43 in the airway epithelium can not only significantly alleviate asthma symptoms, but also resist H1N1 virus infection during asthma attacks. It has very good therapeutic potential for asthma and is expected to provide a new and effective treatment option for asthma and acute asthma exacerbations complicated by respiratory viral infections. Attached Figure Description

[0019] Figure 1 This indicates that asthma modeling can significantly induce the expression level of TDP-43 protein in bronchial epithelial cells, including:

[0020] A shows TDP-43 fluorescent staining in human and mouse lung tissues;

[0021] B represents the TDP-43 immunofluorescence staining and quantitative analysis performed in the lung tissues of mice with chronic obstructive pneumonia and normal mice.

[0022] In C, the left side shows TDP-43 immunofluorescence staining of normal mouse lung tissue in OVA, HDM, and CA-induced asthma model mice; the right side shows TDP-43 protein in Cc10. + Statistical analysis of average fluorescence intensity in cells.

[0023] Figure 2 This indicates that inhibiting TDP-43 expression can suppress the innate immune response in airway epithelial cells, wherein:

[0024] A heatmap shows the differential gene expression between BEAS-2B-si Scramble and BEAS-2B-si TARDBP with and without LPS stimulation; where si TARDBP represents the treatment that inhibits TDP-43 expression and si Scramble represents the control treatment.

[0025] B represents the GO enrichment results of downregulated genes in the TDP-43 knockout group relative to the normal group;

[0026] C is a volcano diagram of differentially expressed genes in control cells (si Scramble) treated with LPS compared to untreated control cells;

[0027] D is a volcano plot of differentially expressed genes in the LPS-treated si TARDBP group relative to the si Scramble group;

[0028] E is a bar graph showing the differential expression of genes involved in different alternative splicing events in the si TARDBP group compared to the si Scramble group treated with LPS. Among them, SE: skipping exon; A5SS: alternative 5' (donor) splicing site; A3SS: alternative 3' (recipient) splicing site; RI: retained intron; MXE: mutually exclusive exon.

[0029] Figure 3 Tardbp ΔScgb1a1 The mice exhibited resistance to an allergen-induced asthma model, including:

[0030] A is Tardbp ΔScgb1a1 TDP-43 immunofluorescence staining of lung tissues from mice and control mice;

[0031] BC was used to compare the Tardbp induced by CA(B) or OVA(C) with H&E staining. ΔScgb1a1 and Tardbp flox Pathological changes among mice;

[0032] DE is a flow cytometry representation showing CA-induced HO-Tardbp. ΔScgb1a1 HE-Tardbp ΔScgb1a1 and Tardbp flox Infiltration of Th2 cells in the lungs of mice (D) and quantitative statistical results (E);

[0033] F stands for Tardbp ΔScgb1a1 and Tardbp flox Flow cytometry results of myeloid cell infiltration in mouse lung tissue;

[0034] G stands for Tardbp ΔScgb1a1 and Tardbp flox Flow cytometry results of immune cell infiltration in mouse BALF;

[0035] H represents serum CBA, as detected by the assay for CA-induced HO-Tardbp. ΔScgb1a1 HE-Tardbp ΔScgb1a1 and Tardbp flox Expression of IL-5 and IFN-γ in mice.

[0036] Figure 4 The Smart-seq2 analysis showed that knockout of TDP-43 in the mouse airway epithelium enhanced selective shearing, where:

[0037] A represents the expression of genes associated with airway hyperresponsiveness (AHR) as shown by a Smart-seq2 heatmap;

[0038] B shows the results of Smart-seq2 gene set enrichment analysis (GSEA) for mRNA detection and selective splicing.

[0039] CD is a heatmap comparison of Tardbp with and without OVA stimulation. ΔScgb1a1 and Tardbp flox Gene expression in mouse airway epithelial cells associated with mRNA surveillance pathway (C) and spliceosome (D);

[0040] E is a stacked histogram showing the Tardbp from OVA stimulation. ΔScgb1a1 and Tardbpflox Changes in alternative splicing events (AS) between airway epithelial cells isolated from mice; SE: skipping exon; A5SS: selective 5' (donor) splicing site; A3SS: selective 3' (recipient) splicing site; RI: intron-retaining; MXE: mutually exclusive exon;

[0041] F, the volcano diagram, illustrates the Tardbp stimulation of OVA. ΔScgb1a1 and Tardbp flox Genes that are differentially expressed among airway epithelial cells of mice, with those marked in red also showing splicing differences.

[0042] Figure 5 This indicates that knocking out TDP-43 in the airway epithelium of mice attenuates the infection of mouse lung tissue by influenza A virus and asthma-associated influenza A virus, among which:

[0043] A represents BEAS-AB cells with TDP-43 knockout and control cells, which were infected with PR8 virus. RT-PCR was used to detect the expression of TDP-43, NP and IFNB1 in the cells.

[0044] B represents the expression levels of NP protein and MX1 in BEAS-AB cells with TDP-43 knockout and control cells after infection with PR8 virus, detected by immunoblotting.

[0045] C is PR8 virus infection Tardbp ΔScgb1a1 and Tardbp flox Mice, statistics on mouse survival;

[0046] DF is a PR8 virus infection of Tardbp ΔScgb1a1 and Tardbp flox Mouse, H&E staining of mouse lung tissue (D), lung injury scoring statistics (E), and representative figures of NP protein immunofluorescence staining results (F).

[0047] GI is for Tardbp ΔScgb1a1 and Tardbp flox A mouse model of asthma induced by OVA combined with PR8 virus infection was established. The results of H&E staining (G), flow cytometry analysis of BALF and immune cell infiltration in lung tissue (H), and immunofluorescence staining of NP protein are represented in Figure I.

[0048] Figure 6 LNP-ASO administration reduces TDP-43 expression in the lungs of mice as a treatment for asthma, wherein:

[0049] A is a representative diagram of the LNP-ASO packaging process;

[0050] BC shows in vivo imaging of various tissues in mice after LNP-ASO was administered via nasal drops, illustrating the drug distribution in tissues (B) and the results of quantitative fluorescence (C).

[0051] D is a flowchart of LNP-ASO treatment in OVA-induced asthmatic mice;

[0052] EG represents the immunohistochemical staining of Tdp43 and Chrm1 in lung tissue of mice treated with LNP-ASO and LNP-NC (E), the flow cytometry results of immune cell infiltration in BALF and lung tissue (F), and the H&E staining representation of lung tissue (G). Detailed Implementation

[0053] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0054] As described in the background section, current first-line clinical medications for asthma (such as glucocorticoids) can lead to drug resistance and serious adverse reactions after long-term use; targeted drugs are mostly focused on targets of downstream type II inflammation (such as IL-4 / IL-4Rα, IL-5 / IL-5R, IL-13, IgE, etc.), which have not solved the problem of asthma recurrence; in addition, there is still a lack of specific targeted therapies for acute asthma exacerbations that are concurrent with respiratory viral infections.

[0055] To address the aforementioned technical problems, this invention conducted extensive experiments and research, and unexpectedly discovered that reducing TDP-43 expression not only significantly alleviates asthma symptoms but also resists H1N1 virus infection during asthma attacks, demonstrating excellent therapeutic potential for asthma.

[0056] The following will be combined with the appendix Figure 1 To be continued Figure 6 The experimental investigation process of the present invention is described in detail, along with specific embodiments.

[0057] Example 1: Investigating the mechanism by which inhibiting TDP-43 expression can improve asthma symptoms.

[0058] Research suggests that the airway epithelium is the host's first line of defense against allergen exposure and the initial site of airway hyperresponsiveness. Therefore, identifying key host factors in airway epithelial cells that trigger airway hyperresponsiveness, and providing new targets for targeted therapy of asthma, is a pressing issue in this field. To this end, this invention has conducted extensive research:

[0059] 1. Asthma modeling can significantly induce the expression level of TDP-43 protein in bronchial epithelial cells.

[0060] First, in the preliminary research of this invention, it was found that TDP-43 is specifically expressed in lung epithelial cells in lung tissue ( Figure 1 (A) It was also found that the expression level of TDP-43 in the lung mucosa of patients with chronic obstructive pulmonary disease (COPD) was significantly higher than that in normal individuals. Figure 1 Based on this finding, this invention induced various COPD mouse models using ovalbumin (OVA), cockroach antigen (CA), or house dust mite (HDM). In the lung mucosa of each model mouse, the expression level of TDP-43 was still significantly higher than that of the control group mice. Figure 1 These results suggest that TDP-43 may play an important role in the occurrence and development of chronic obstructive pulmonary disease (COPD).

[0061] 2. Down-knock TDP-43 can significantly inhibit the inflammatory response of airway cells.

[0062] Based on the above findings, in order to explore the potential role of TDP-43 protein in airway inflammatory response, this invention knocked down the expression of TDP-43 in human airway epithelial cells BEAS-2B using si TARDBP, and included si Scramble as a control treatment; subsequently, the knocked-down cells and control cells were treated with LPS (lipopolysaccharide) to induce inflammation, and PBS was included as a control treatment.

[0063] Full-length transcriptome sequencing (Smart-seq2) of the treated cells revealed that LPS stimulation significantly induced the expression of genes related to innate immune responses, accompanied by an increase in alternative splicing events. Figure 2 (AC), while knocking down TDP-43 expression can significantly inhibit the expression of genes related to innate immune response (AC). Figure 2 (D), while the occurrence of selective shear events is significantly increased ( Figure 2 These results indicate that knocking down TDP-43 can significantly suppress the inflammatory response of airway cells.

[0064] 3. Knocking out TDP-43 expression significantly attenuated the development and progression of allergen-induced asthma in mice.

[0065] This embodiment uses the CRISPR / Cas gene editing system to target and knock out TDP-43, constructing a Tardbp mouse model with airway epithelium-specific TDP-43 knockout. ΔScgb1a1 Simultaneously, littermates of flox mice that did not undergo TDP-43 knockout were included as control mice (Tardbp). flox Further, using cockroach antigen (CA) to induce Tardbp ΔScgb1a1 Mice and Tardbp floxThe development of asthma in mice was induced to evaluate the therapeutic effect of TDP-43 in asthma. The CRISPR / Cas gene editing system includes gRNA1 and gRNA2 that specifically target the TDP-43 encoding gene. The sequence of gRNA1 is 5'-ATTTCTTAGTGTATGTCCAT-3' (SEQ ID NO: 1), and the sequence of gRNA2 is 5'-TGGGAGTGGGGACCCTGCCT-3' (SEQ ID NO: 2). Both recognize the PAM sequence GGG.

[0066] The results showed that, compared with the control group mice (Tardbp) flox TDP-43 homozygous knockout mice (HO-Tardbp) ΔScgb1a1 ) and TDP-43 heterozygous knockout mice (HE-Tardbp) ΔScgb1a1 The asthma phenotype of all mice was significantly reduced, and the asthma phenotype of homozygous TDP-43 knockout mice was even weaker than that of heterozygous knockout mice. Figure 3 (AB). This embodiment also used ovalbumin (OVA) to induce asthma in TDP-43 knockout mice and control mice. The results also showed that TDP-43 knockout significantly attenuated the asthma phenotype in mice. Figure 3 (C).

[0067] Flow cytometry results of lung tissue showed that, compared to the control group mice (Tardbp) flox TDP-43 knockout mice (HO-Tardbp) ΔScgb1a1 HE-Tardbp ΔScgb1a1 IL-4 + CD4 + T cells, IL-5 + CD4 + T cells, IL-13 + CD4 + T cell infiltration was significantly reduced. Figure 3 DE), while TDP-43 knockout mice (HO-Tardbp) ΔScgb1a1 The infiltration of eosinophils, neutrophils, and macrophages in the mice was also significantly lower than in the control group mice. Figure 3 Flow cytometry analysis of bronchoalveolar lavage fluid (BALF) in mice showed that TDP-43 knockout mice (HO-Tardbp) ΔScgb1a1 The infiltration of eosinophils, neutrophils, and macrophages in the bronchoalveolar lavage fluid of mice was significantly lower than that in the control group. Figure 3 (G). HO-Tardbp with asthma ΔScgb1a1 Mice and HE-Tardbp ΔScgb1a1Decreased concentrations of IL-5 and IFN-γ were also observed in BALF of mice. Figure 3 H).

[0068] The above results indicate that allergen-induced TDP-43 expression plays a pathogenic role in the development of asthma, and knocking out TDP-43 expression can significantly reduce the occurrence and development of allergen-induced asthma in mice.

[0069] 4. Exploring the mechanism by which inhibiting TDP-43 can alleviate the occurrence of asthma.

[0070] To explore the mechanism by which TDP-43 alleviates the development and progression of asthma, this embodiment first constructed a homozygous TDP-43 conditional knockout mouse (Tardbp). ΔScgb1a1 After inducing asthma in mice using ovalbumin (OVA), Tardbp treated with OVA was isolated. ΔScgb1a1 Mice and control mice (Tardbp) flox The airway epithelial cells were analyzed and full-length transcriptome sequencing (Smart-seq2) was performed.

[0071] Sequencing results showed that control mice treated with ovalbumin exhibited a strong transcriptomic profile characteristic of airway hyperresponsiveness, while airway epithelial cells from TDP-43 conditional knockout mice that received the same induction did not show the characteristics of airway hyperresponsiveness. Figure 4 (A).

[0072] Collect homozygous TDP-43 conditional knockout mice (Tardbp) ΔScgb1a1 Compared to the control group mice (Tardbp) flox GSEA enrichment analysis of differentially expressed genes showed that mRNA detection and alternative splicing signaling pathways were significantly enriched. Figure 4 B), and Tardbp ΔScgb1a1 The mRNA levels and expression of alternative splicing-related genes in mice were significantly higher than those in the control group. Figure 4 The results of the stacked histogram also showed that, compared with the control group, Tardbp from the OVA-treated group... ΔScgb1a1 Selective shearing events were significantly altered in mouse airway epithelial cells, and the expression of genes related to airway hyperresponsiveness was significantly reduced. Figure 4 (E). Detailed analysis results showed that alternative splicing of genes such as Chrm1, Noxa1, Abcc8, Ecm1, Slc28a3, and Pdzk1ip1 occurred in Tardbp. ΔScgb1a1 The group significantly increased, and gene expression significantly decreased. Figure 4 (F).

[0073] These results indicate that inhibiting TDP-43 expression may suppress the inflammatory response of airway cells by regulating the occurrence of selective shearing events, thereby alleviating the development of asthma in mice.

[0074] Example 2: Inhibiting TDP-43 expression can enhance resistance to influenza A virus infection.

[0075] Asthma patients co-infected with influenza virus pose a significant challenge to their survival. This invention aims to investigate the effect of TDP-43 knockout on the ability of airway epithelial cells to resist influenza virus.

[0076] TDP-43 knockout BEAS-2B cells (denoted as sgTARDBP) and control cells (denoted as sgScramble) were treated with / without PR8 strain (IAV) infection. Immunoblotting was used to detect the expression levels of NP protein and MX1 in the cells, and RT-PCR was used to detect the expression levels of TDP-43, NP, and IFNB1. The ribonucleoprotein complex formed by NP protein is essential for viral transcription and replication, and its expression level indicates viral load; MX1 protein, as a key effector molecule in the host's IFN response, reflects the strength of the host's antiviral defense; IFNB1 contributes to resistance to viral infection. The results showed that TDP-43 knockout BEAS-2B cells were significantly insensitive to PR8 strain infection. Figure 5 (AB).

[0077] It also involved nasal administration to homozygous TDP-43 conditional knockout mice (HO-Tardbp). ΔScgb1a1 ) and control mice (Tardbp) flox Following 8 consecutive days of infection with influenza A virus (H1N1, PR8 strain), the following procedures were performed: survival status was assessed, H&E staining analysis of lung tissue was conducted, lung injury scores were calculated, and immunofluorescence staining analysis of NP protein was performed. Results showed that, compared to Tardbp... flox Mice, pure HO-Tardbp ΔScgb1a1 The survival rate of mice infected with IAV (Influenza A Virus) was significantly increased. Figure 5 (C). Furthermore, H&E staining results and lung injury scoring statistics showed that HO-Tardbp ΔScgb1a1 The lung lesions were significantly milder. Figure 5 (DE). Immunofluorescence staining results also showed HO-Tardbp. ΔScgb1a1 The number of viral particles in the lungs of mice was significantly lower than that in the control group mice. Figure 5 (F).

[0078] Furthermore, this was also achieved by using TDP-43 conditional knockout mice (HO-Tardbp) ΔScgb1a1 ) and control mice (Tardbp) flox Eight days after OVA stimulation combined with influenza A virus (IVA) infection, mice underwent the following treatments: H&E staining of lung tissue, flow cytometry analysis of BALF and immune cell infiltration in lung tissue, and immunofluorescence staining analysis of NP protein. Results showed that, compared to the control group, HO-Tardbp... ΔScgb1a1 The lung lesions in the group of mice were significantly reduced. Figure 5 The number of viral particles in the lungs was also significantly reduced (GH). Figure 5 GI).

[0079] The above results indicate that inhibiting TDP-43 expression can enhance resistance to H1N1 influenza virus, reduce H1N1 infection, and thus effectively prevent the aggravation of asthma symptoms.

[0080] Example 3: Treatment of Asthma with ASO that Targets and Inhibits TDP-43 Expression

[0081] Based on the observed significant phenotype of TDP-43 knockout in reducing the occurrence of asthma and weakening H1N1 virus infection in mice, thus preventing the exacerbation of asthma symptoms, this invention aims to find a drug that can directly target TDP-43 and inhibit its expression in order to effectively treat asthma and the exacerbation of asthma symptoms caused by H1N1 virus infection.

[0082] Antisense oligonucleotides (ASOs) are a gene therapy-based approach. ASOs utilize short single-stranded nucleic acid molecules that complement specific mRNA sequences to form stable double-stranded structures. This recruits RNase H to cleave target mRNA or regulates alternative splicing and protein translation inhibition of target genes through steric hindrance, thereby inhibiting the production of disease-related proteins or altering their function. ASOs are relatively easy to design and synthesize, and their key advantages are precise targeting of disease-related genes, high specificity, and few adverse reactions. However, the instability and immunogenicity of ASOs limit their clinical application. Lipid nanoparticles (LNPs) can effectively overcome these limitations and have become a key technology for nucleic acid drug delivery. Therefore, this embodiment selects the ASO-LNP strategy to design a candidate drug targeting TDP-43 to inhibit asthma progression.

[0083] In this embodiment, two ASO sequences for interfering with TDP-43 were designed, namely ASO-1 and ASO-2, and the specific sequences are as follows:

[0084] The ASO-1 nucleotide sequence is: 5'-TTTCAATGGGTTCATCGTTC-3' (SEQ ID NO: 3);

[0085] The ASO-2 nucleotide sequence is: 5'-TACACTGAGACACGGGATTC-3' (SEQ ID NO: 4).

[0086] The two ASO sequences were respectively LNP-packaged, denoted as LNP-ASO-1 and LNP-ASO-2. The morphology of LNP-ASO-1 was observed using electron microscopy. Figure 6 (A). Transfection of B16-F10 cells with LNP-ASO-1 and LNP-ASO-2 significantly inhibited TDP-43 expression. Immunofluorescence observation of drug distribution in mice after intranasal administration of LNP-ASO showed that intranasal administration of LNP-ASO had good lung delivery specificity. Figure 6 (BC).

[0087] Furthermore, during the modeling process, OVA-induced asthmatic mice were treated with LNP-ASO-1 via nasal drops every 4 days. Figure 6 The study included a control group (LNP-NC, where the LNP vector includes an NC sequence that does not target any gene). After treatment, the following steps were performed: immunohistochemical analysis of Tdp43 and Chrm1 in mouse lung tissue, flow cytometry analysis of immune cell infiltration in BALF and lung tissue, and H&E staining analysis of lung tissue. Results showed that the expression of Tdp43 and Chrm1 in the lung tissue of mice treated with ASO (LNP-ASO-1) was significantly reduced. Furthermore, the infiltration of immune cells in BALF and lung tissue in the treatment group was significantly lower than that in the control group (LNP-NC). Figure 6 The F), and the lung lesions in the treatment group mice were significantly milder (F). Figure 6 (G).

[0088] In summary, this invention has for the first time discovered the role of TDP-43 in improving asthma and H1N1 virus infection through experiments. Experimental verification shows that inhibiting TDP-43 expression can not only significantly alleviate asthma symptoms, but also resist H1N1 virus infection during asthma attacks. It has great potential for the treatment of asthma and is expected to provide a new and effective treatment option for asthma and acute asthma exacerbations complicated by respiratory viral infections.

[0089] Although the present invention has been described in detail through the preferred embodiments above, it should be understood that the above description should not be considered as a limitation of the present invention. Various modifications and substitutions to the present invention will be apparent to those skilled in the art after reading the above description. Therefore, the scope of protection of the present invention should be defined by the appended claims.

Claims

1. Application of TDP-43 inhibitors in the preparation of drugs for the prevention and / or treatment of asthma.

2. The application as described in claim 1, characterized in that, The asthma mentioned includes asthma exacerbations caused by respiratory viral infections.

3. The application as described in claim 1 or 2, characterized in that, The TDP-43 inhibitor is a substance that knocks out or knocks down the gene encoding TDP-43, or reduces the expression level or activity of TDP-43 protein, or weakens the effect of TDP-43.

4. The application as described in claim 3, characterized in that, The substance includes any one of the following: protein, nucleic acid molecule, small molecule compound, or gene intervention tool capable of knocking out or knocking down TDP-43 expression.

5. Application of TDP-43 inhibitors in the preparation of drugs for the prevention and / or treatment of influenza A infection.

6. An antisense oligonucleotide targeting TDP-43, characterized in that, The sequence of the antisense oligonucleotide is shown in SEQ ID NO: 3 or SEQ ID NO:

4.

7. The antisense oligonucleotide as described in claim 6, characterized in that, The antisense oligonucleotides are delivered via lipid nanoparticle carriers.

8. A CRISPR / Cas gene editing system for targeting and knocking out the coding gene of TDP-43, characterized in that, The gene editing system includes a gRNA that specifically targets the gene encoding TDP-43, the sequences of which include SEQ ID NO: 1 and SEQ ID NO:

2.

9. The use of the antisense oligonucleotide of claim 6 or 7, or the gene editing system of claim 8, in the preparation of a medicament for the prevention and / or treatment of asthma or a medicament for the treatment of influenza A infection.

10. The application as described in claim 9, characterized in that, The dosage forms of the drug include nebulizers and drops.

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