Method for predicting clinical outcome of treatment of CRSwNP patient by omaszumab according to baseline expression level of cytokines in nasal secretions and application of clinical outcome of treatment of CRSwNP patient by omaszumab

Abstract

The invention discloses a method for predicting the clinical outcome of treatment of a CRSwNP patient by omphizumab according to the baseline expression level of cytokines in nasal secretions and application of the method. 48 cytokines in the nasal secretions of the patient are detected before treatment by omphizumab; the clinical outcome of nasal polyp improvement of a CRSwNP patient treated by omaszumab can be noninvasively predicted by virtue of the baseline expression level of any one of seven cell factors including Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO and IgE in nasal secretions of the patient, wherein the NPS improvement of the treatment improvement group and the poor treatment group is greater than that of the treatment improvement group and the poor treatment group is found for the first time. The clinical outcome of the nasal polyp improvement of the CRSwNP patient treated by omaszumab can be noninvasively predicted.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a method for predicting the clinical outcome of CRSwNP patients treated with omalizumab based on the baseline expression level of cytokines in nasal secretions and its application. Background Art

[0002] Chronic rhinosinusitis (CRS) is a common chronic inflammatory disease involving the sinus mucosa in clinical practice. The course of the disease is more than 12 weeks, and the clinical symptoms are mostly nasal congestion, runny nose, facial pain, hyposmia or anosmia.

[0003] CRS can be divided into two types based on clinical phenotype: chronic rhinosinusitis with nasal polyps (CRSwNP) and chronic rhinosinusitis without nasal polyps (CRSsNP). [2] Although the phenotypic classification of CRS has been established, both CRS can be further divided into different endotypes based on the underlying immune type and cell differentiation pattern. [3] .

[0004] In general, CRSwNP patients have more severe clinical symptoms, a relatively high recurrence rate, and a high comorbidity rate of diseases such as bronchial asthma, aspirin-exacerbated respiratory diseases, and allergic rhinitis. From the perspective of immune endotype, CRS can be divided into type 1 (type 1) represented by interferon-(IFN-), type 2 (type 2) represented by IL-4, IL-5, and IL-13, and type 3 (type 3) CRS represented by the IL-17 family, according to different inflammatory factor driving modes. CRSsNP is usually dominated by type 1 and type 3 inflammation, and CRSwNP is dominated by eosinophil infiltration and type 2 inflammation. [4] Although the “one-to-two” phenotypic classification method is still generally applicable to clinical treatment, the endotype study of CRS reflects the complexity of the pathogenesis of CRS, especially the mixed inflammatory pattern that often occurs in refractory CRS. [5-7] Type 2 CRS plays a decisive role in the patient's quality of life, disease severity, and disease prognosis [8] .

[0005] Omalizumab is a biologic agent for type 2 CRS, but its clinical outcomes in patients with CRSwNP vary [9] .

[0006] The Chinese patent application number 202311274974.4, "A method and application for predicting or evaluating the efficacy of omalizumab in the treatment of recurrent nasal polyps using local IgE", discloses that: the high expression level of total IgE in nasal secretions may predict poor clinical outcomes in patients with CRSwNP treated with omalizumab. ① According to the EPOS2020 guidelines and EUFOREA's recommendations, the first evaluation time for biologics is 6 months after treatment. The evaluation time after omalizumab treatment in this patent is 16 weeks. The short evaluation time may lead to inaccurate evaluation results; ② This patent explores whether the IgE+ cell level in nasal polyp tissue can be used as a predictor for predicting the efficacy of omalizumab treatment, which is an invasive operation; ③ The processing of nasal secretions also requires centrifugation and standardized quantitative processing of BCA, and judgment after immunostaining, which is more complicated; ④ In addition, the expression of different inflammatory factors in patients varies greatly, and the expression level of only one cytokine may not be enough to fully explain the patient's disease level.

[0007] Research progress on omalizumab in the treatment of chronic rhinosinusitis with nasal polyps

[10] Reported: Mortuaire et al. studied and analyzed the eosinophil counts, IgE, IL-5, IgA, ECP and other factors in the blood, nasal secretions and nasal polyps of 57 patients with CRSwNP. Except for the serum total IgE level in the allergic group, which was higher than that in the control group, there were no significant differences in other levels. This article is a general review of the effectiveness of omalizumab in the treatment of chronic sinusitis with nasal polyps, and did not propose its potential effective indicators for predicting clinical prognosis.

[0008] Research progress on omalizumab targeted therapy for chronic sinusitis with nasal polyps

[11] It was reported that in CRSwNP, the epithelial cells (ECs) of the sinuses are the first line of defense to protect the nasal cavity. In nasal polyps, ECs can trigger a type 2 inflammatory response, and the epithelial barrier of the nasal and sinonasal mucosa is destroyed and lacks structural integrity, resulting in an active epithelial cell inflammatory state. ECs can produce IL-33, IL-1, thymic stromal lymphopoietin (TSLP), and IL-25 to respond to various damage-related substances, such as pathogenic microorganisms and their related products or allergens. Superantigens from the nasal cavity, Staphylococcus aureus colonies, can lead to the activation of polyclonal T cells and B cells, thereby amplifying the inflammatory response. Epithelial cytokines such as TSLP, IL-25, and IL-33 induce the activation and accumulation of type 2 innate lymphocytes (ILC2), leading to the production of IL-5, IL-13, and IL-4. At the same time, helper T cell 2 (Th2) cells can directly respond to IL-25 and IL-33, further amplifying the inflammatory response. This article is a general review of the effectiveness of omalizumab in the treatment of chronic sinusitis with nasal polyps, and does not propose its potential effective indicators for predicting clinical prognosis.

[0009] Currently, there is no non-invasive method to predict the clinical outcomes of patients with chronic sinusitis and nasal polyps treated with omalizumab. Establishing a method to predict the clinical outcomes of patients with chronic sinusitis and nasal polyps treated with omalizumab is of great significance in guiding the clinical use of omalizumab.

[0010] References:

[0011] [1] Bachert C, Marple B, Schlosser RJ, et al. Adult chronicrhinosinusitis. Nat Rev Dis Primers. 2020; 6(1):86.

[0012] [2]Shi JB, Fu QL, Zhang H, et al. Epidemiology of chronic rhinosinusitis: results from a cross-sectional survey in seven Chinese cities. Allergy.

[0013] 2015;70(5):533-539.

[0014] [3]Klingler AI,Stevens WW,Tan BK,et al.Mechanisms and biomarkers ofinflammatory endotypes in chronic rhinosinusitis without nasal polyps.JAllergy Clin Immunol.2021;147(4):1306-1317.

[0015] [4]Bachert C,ZhangN,Hellings PW,Bousquet J.Endotype-driven carepathways in patients with chronic rhinosinusitis.J Allergy Clin Immunol.2018;141(5):1543-1551.

[0016] [5]Cho SH,Kim DW,Gevaert P.Chronic Rhinosinusitis without NasalPolyps.J Allergy Clin Immunol Pract.2016;4(4):575-582.

[0017] [6]Tomassen P,Vandeplas G,Van Zele T,et al.Inflammatory endotypes ofchronic rhinosinusitis based on cluster analysis of biomarkers.J Allergy ClinImmunol.2016;137(5):1449-1456.e1444.

[0018] [7]Kato A,Schleimer RP,Bleier BS.Mechanisms and pathogenesis ofchronic rhinosinusitis.J Allergy Clin Immunol.2022;149(5):1491-1503.

[0019] [8]Wang

[0020] [9] Fokkens WJ, Lund VJ, Hopkins C, et al. European Position Paper on Rhinosinusitis and Nasal Polyps 2020. Rhinology. 2020; 58 (Suppl S29): 1-464.

[0021]

[10] Wang Aowei, Shi Wenjie. Research progress of omalizumab in the treatment of chronic rhinosinusitis with nasal polyps[J]. International Journal of Otorhinolaryngology Head and Neck Surgery, 2021, (Issue 4).

[0022]

[11] Zhang Yunqiu, Ren Xiumin, Xu Ou, Dong Jinhui, Wang Jianxing. Research progress of omalizumab targeted therapy for chronic sinusitis with nasal polyps [J / OL]. Journal of Otolaryngology and Ophthalmology of Shandong University. https: / link.cnki.net / urlid / 37.1437.R.20240408.1519.006 Summary of the invention

[0023] In view of the fact that there is no non-invasive method for predicting the clinical outcome of patients with chronic sinusitis and nasal polyps treated with omalizumab at this stage, the present application detects 48 cytokines in the nasal secretions of patients before omalizumab treatment, and finds that the baseline expression levels of seven cytokines, Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in the nasal secretions of patients can predict the clinical outcome of nasal polyp improvement in patients with CRSwNP after 24 weeks of omalizumab treatment. To achieve the above purpose, the specific technical scheme of the present invention is as follows:

[0024] First, basic patient information and research design

[0025] A total of 18 patients with chronic sinusitis and nasal polyps aged 18 years and above participated in this study. The basic characteristics of the patients were collected, including gender, age, BMI, asthma, allergic rhinitis, nasal polyps VAS score, nasal congestion VAS score, runny nose VAS score, olfaction VAS score, headache VAS score, SNOT-22 score, and peripheral blood eosinophil count.

[0026] Patients receiving omalizumab treatment will receive the required subcutaneous injection dose according to the patient's baseline body weight (kg) and serum total IgE (kU / L) in accordance with the instructions.

[0027] Second, non-invasive sampling to detect the expression levels of 48 baseline cytokines in patients’ nasal secretions

[0028] Before omalizumab treatment, non-invasive sampling was performed to detect the baseline expression levels of 48 cytokines in the patients' nasal secretions; at 24 weeks of treatment, non-invasive sampling was performed again to detect the expression levels of 48 cytokines in the patients' nasal secretions.

[0029] 44 cytokines including IL-1α, IL-1β, IL-1Rα, and IL-2 were detected by Luminex;

[0030] MPO and periostin in serum were detected by enzyme-linked immunosorbent assay (Elisa);

[0031] The expression levels of eosinophil cationic protein (ECP) and total IgE were automatically detected by the ImmunoCAP system.

[0032] The third aspect is the evaluation index of clinical symptom improvement

[0033] The NPS scores of the enrolled patients were collected at baseline and at 4, 8, 12, 16, 20, and 24 weeks after the administration of omalizumab. An improvement of more than 2 points in the NPS score was considered treatment improvement, and the treated patients were divided into a treatment improvement group and a poor treatment group. At 24 weeks, among the 18 patients, 10 had an NPS improvement of more than 2 points, accounting for 55.56%.

[0034] Fourthly, differentially expressed cytokines and predictive efficacy in nasal secretions of patients in the treatment improvement group and the poor treatment group

[0035] Results: The baseline expression levels of Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in nasal secretions predicted the clinical outcomes of omalizumab treatment for improvement of nasal polyps in patients with CRSwNP with ROC-AUCs of 0.788, 0.813, 0.812, 0.800, 0.788, 0.781, and 0.900, respectively.

[0036] Fifth aspect: Verify the predictive efficacy of the seven cytokines in the fourth aspect

[0037] Results: The baseline expression levels of Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in nasal secretions had good accuracy, sensitivity, and specificity in predicting the clinical outcomes of nasal polyp improvement in patients with CRSwNP treated with omalizumab, and can be used to predict the clinical outcomes of patients.

[0038] Compared with the prior art, the beneficial effects of this application are:

[0039] For the first time, a method was established to non-invasively predict the clinical outcomes of omalizumab-treated patients with CRSwNP using baseline expression levels of Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in nasal secretions. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 , differentially expressed cytokines in nasal secretions of CRSwNP patients treated with omalizumab, the treatment improvement group and the poor treatment group divided by NPS improvement greater than 2 points,

[0041] A is a statistical diagram of the difference in the expression of Eotaxin in nasal secretions between the treatment improvement group and the poor treatment group.

[0042] B is a statistical diagram of the difference in expression of GM-CSF in nasal secretions between the treatment improvement group and the poor treatment group.

[0043] C is a statistical diagram of the difference in expression of CXCL-1 in nasal secretions between the treatment improvement group and the poor treatment group.

[0044] D is the statistical diagram of the difference in expression of IP-10 in nasal secretions between the treatment improvement group and the poor treatment group.

[0045] E is a statistical diagram showing the difference in expression of IL-1a in nasal secretions between the treatment improvement group and the poor treatment group, F is a statistical diagram showing the difference in expression of MPO in nasal secretions between the treatment improvement group and the poor treatment group, and G is a statistical diagram showing the difference in expression of IgE in nasal secretions between the treatment improvement group and the poor treatment group;

[0046] Figure 2 , ROC curve of 7 cytokines in nasal secretions predicting the clinical outcomes of improvement of nasal polyps in patients with CRSwNP treated with omalizumab.

[0047] Figure 1 middle,

[0048] *, **, *** and **** represent P<0.05, P<0.01, P<0.001 and P<0.0001, respectively, indicating that the difference between the two groups was statistically significant. DETAILED DESCRIPTION

[0049] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

[0050] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods.

[0051] All materials, reagents, etc. in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0052] Reagents and Materials

[0053]

[0054]

[0055]

[0056] instrument

[0057]

[0058] Example 1. Patient basic information and study design

[0059] This study was a single-center trial conducted at Beijing Tongren Hospital from January 2019 to January 2023.

[0060] A total of 18 patients with chronic sinusitis and nasal polyps aged 18 years and above and 16 patients aged 18 years and above who required nasal expansion surgery due to nasal septum deviation participated in this study.

[0061] This study was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (TRECKY2019-070). Written informed consent was obtained before patient participation.

[0062] Patients receiving omalizumab treatment will be given the required subcutaneous injection dose according to the patient's body weight (kg) and serum total IgE (kU / L) at baseline according to the standards of the instructions. During the observation period, patients are allowed to use nasal spray glucocorticoids, antihistamines and antihistamines to treat nasal diseases, and are advised to use bronchial asthma treatment drugs regularly. The basic characteristics of the patients are shown in the table below.

[0063]

[0064]

[0065] Example 2. Evaluation indicators for improvement of clinical symptoms

[0066] Demographic data were collected for all patients. Nasal polyp scores (NPS) under nasal endoscopy were collected at baseline, 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, and 24 weeks for further analysis.

[0067] NPS scores range from 0 to 8, where 0 = no polyps, 1 = small polyps visible in the middle nasal passages, no larger than the lower edge of the middle turbinate, 2 = polyps reaching the lower edge of the middle turbinate, 3 = polyps reaching the lower edge of the inferior turbinate, or polyps with a score of 2 accompanied by large polyps on the inner side of the middle turbinate, and 4 = polyps completely or nearly completely blocking the nasal cavity, i.e., to the floor of the nose

[0068] An improvement of more than 2 points in NPS was considered treatment improvement. At 24 weeks, the treated patients were divided into a treatment improvement group and a poor treatment group. Among the 18 patients, 10 had an NPS score improvement of more than 2 points, accounting for 55.56%.

[0069] Example 3: Detection of baseline expression levels of 48 cytokines in nasal secretions of patients

[0070] 1. Sampling

[0071] Samples were collected before and after 24 weeks of omalizumab treatment.

[0072] Prepare 2 strips of expansion sponges suitable for the size of the nasal cavity in advance in a 15mL centrifuge tube and weigh them for later use. When using, use a gun-shaped forceps to take out the expansion sponge cotton strips and put them into the left and right nasal passages of the patient for about 5 to 10 minutes respectively to collect nasal secretions. After the sponge absorbs the nasal secretions until it is fully expanded, take it out and put it back into the original 15mL centrifuge tube, tighten the centrifuge tube cap and weigh it again. Add 3mL of normal saline and let it stand in a 4℃ refrigerator for 2h; place the sponge in a 5mL syringe, put it into the original 15mL centrifuge tube, centrifuge it at 1500g for 15min, take out the lower layer of liquid and store it in a 1.5mL transparent sterile centrifuge tube, and store it in a -80℃ refrigerator.

[0073] 2. Detection

[0074] 2.1 Luminex detection

[0075] The expression levels of 44 cytokines in the serum of patients at baseline and after 24 weeks of omalizumab treatment were detected by Luminex, including interleukin (IL)-1α, IL-1β, IL-1Rα, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-15, IL-17, IL-25, IL-33, CD40L, epidermal growth factor (EGF), eosinophil chemotactic factor (Eotaxin), basic fibroblast growth factor (FGF-basic), FMS-like tyrosine kinase receptor 3 ligand (Flt-3 Ligand), granulocyte colony-stimulating factor (GFS), and pro-inflammatory cytokine (PFS). factor, G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), granzyme B, chemokine ligand-1 (CXC motif, CXCL-1), CXCL-2, interferon (interferon, IFN)-α, IFN-β, IFN-γ, IP-10, chemokine (C-C motif) ligand (CCL)-2, CCL-3, CCL-4, CCL-5, CCL-19, CCL-20, programmed death receptor-1 (cell death ligand 1, PD-L1), platelet-derived growth factor (PDGF)-AA, PDGF-AB, transforming growth factor (TGF)-α, tumor necrosis factor (TNF)-α, tumor necrosis factor-related apoptosis-inducing ligand (TNF-related The main inhibitors of TNF-α and TNF-α are TRAIL, apoptosis-inducing ligand and vascular endothelial growth factor (VEGF).

[0076] (1) Before the experiment, restore the Human XL Cytokine Fixed Panel (RD#LKTM04 and RD#LKTM014B) kit to room temperature; prepare the standard, washing solution, quality control, mixed microsphere solution, Biotin-antibody solution and PE-streptavidin solution according to the instructions;

[0077] (2) Prepare the plate layout and add 50 μL of standard or sample to be tested to each well;

[0078] (3) Add 50 μL of resuspended magnetic beads to each well, seal the plate with aluminum foil, incubate at room temperature for 2 h, and adjust the shaking speed to 800 rpm;

[0079] (4) Add the magnetic plate to the bottom of the microplate, let it stand for 1 min, discard the liquid, take the magnetic plate, add 100 μL of washing solution to each well, let it stand for 1 min, add the magnetic plate to the bottom of the microplate again, let it stand for 1 min, and discard the liquid. Repeat the above washing steps 3 times;

[0080] (5) Add 50 μL of Biotin-antibody to each well, seal the plate with aluminum foil, incubate at room temperature for 1 h, and adjust the shaking speed to 800 rpm;

[0081] (6) Add the magnetic plate to the bottom of the microplate, let it stand for 1 min, discard the liquid, take the magnetic plate, add 100 μL of washing solution to each well, let it stand for 1 min, add the magnetic plate to the bottom of the microplate again, let it stand for 1 minute, and discard the liquid. Repeat the above washing steps 3 times;

[0082] (7) Add 50 μL of PE-streptavidin to each well, seal the plate with aluminum foil, incubate at room temperature for 30 min, and adjust the shaking speed to 800 rpm;

[0083] (8) Add the magnetic plate to the bottom of the microplate, let it stand for 1 min, discard the liquid, take the magnetic plate, add 100 μL of washing solution to each well, let it stand for 1 min, add the magnetic plate to the bottom of the microplate again, let it stand for 1 min, and discard the liquid. Repeat the above washing steps 3 times;

[0084] (9) Add 100 μL of washing solution to each well and shake at 800 rpm for 2 min at room temperature to suspend the magnetic beads;

[0085] (10) Read the results on a Luminex 200 instrument within 90 min.

[0086] 2.2 Enzyme-linked immunosorbent assay (ELISA) detection

[0087] Serum MPO and periostin were detected by enzyme-linked immunosorbent assay (ELISA).

[0088] (1) Before the experiment, equilibrate the Elisa kit to room temperature according to the instructions, prepare the standard and washing solution according to the instructions, and prepare the nasal secretions and serum samples to be tested;

[0089] (2) Prepare the plate layout and add 100 μL of RD1-27 to each well;

[0090] (3) Within 15 min, add 50 μL of standard or sample to be tested to each well, seal the plate, and incubate at room temperature for 2 h with the shaker set to 500 rpm;

[0091] (4) Remove the liquid in the wells, add 400 μL of washing solution to each well, soak for 1 to 2 min, remove and tap the liquid in the wells, and wash the plate 4 times in total;

[0092] (5) Add 200 μL of MPO conjugate to each well, seal the plate, incubate at room temperature for 2 h, and adjust the shaking speed to 500 rpm;

[0093] (6) Remove the liquid in the wells, add 400 μL of washing solution to each well, soak for 1 to 2 hours, remove and tap the liquid in the wells, and wash the plate 4 times in total;

[0094] (7) Add 200 μL of substrate solution to each well, seal the plate, and incubate at room temperature in the dark for 30 min;

[0095] (8) Add 50 μL of stop solution to each well and gently shake to ensure thorough mixing;

[0096] (9) Immediately measure the OD450-540 values ​​using a microplate reader;

[0097] (10) Calculate the protein concentration in the sample based on the standard curve.

[0098] 2.3 ImmunoCAP Phadiatop detection

[0099] The expression levels of eosinophil cationic protein (ECP) and total IgE were automatically detected by the ImmunoCAP system.

[0100] Turn on the Phadia 1000 instrument system and Prime software, and the instrument automatically enters the ISW software interface. Click "LOAD REAGENTS" in the ISW software, add and replenish relevant reagents and consumables (Wash Buffer, substrate solution, stop solution, enzyme-labeled secondary antibody, curve control or calibration products, etc.) according to the instrument menu, start the instrument for detection, and check the result data after the detection is completed.

[0101] 3. Statistical methods

[0102] Data analysis was performed using IBM SPSS 23.0 software (SPSS, USA) and GraphPad Prism 9 software (GraphPad Software, USA). Baseline characteristics of patients were expressed as mean ± SD or median-interquartile range (IQR) for continuous variables; Kolmogorov-Smirnov test was used to assess the normal distribution of continuous variable data; Fisher exact test or Chi-squared test was used to compare gender, residential area, education level, smoking status, comorbidities, and monthly income between the two groups. Mann-Whitney Utest was used for intergroup comparison of continuous variables, that is, to compare age, BMI, and cytokine expression levels between the two groups. Wilcoxon signed rank test was used for paired sample test, and Bonferroni correction was used for multiple testing. Spearman correlation analysis was used to evaluate the correlation between biomarkers and the degree of improvement of clinical symptoms, and the BH (Benjamini-Hochberg) method was used for multiple testing correction. ROC curve (Receiveroperating characteristic curve) was used to evaluate the ability of different clinical or cytokine factors to predict the response to omalizumab treatment of CRSwNP. Youden index determined the optimal cutoff value and was used to calculate sensitivity and specificity. P < 0.05 was considered statistically significant.

[0103] Example 4: Differentially expressed cytokines in nasal secretions of patients in the treatment improvement group and the poor treatment group

[0104] 1. Baseline expression levels of 48 cytokines in nasal secretions of patients in the treatment improvement group and the poor treatment group

[0105] The baseline expression levels of 48 cytokines in the patients' nasal secretions were detected. According to the improvement of the patients' clinical symptoms after treatment, the NPS score was improved by more than 2 points, and the patients were divided into a treatment improvement group and a poor treatment group. The differentially expressed cytokines were analyzed. The results are shown in Table 1.

[0106] Table 1. Baseline cytokine expression levels of CRSwNP patients grouped according to improvement in NPS score of more than 2 points

[0107]

[0108]

[0109]

[0110] 2. Differentially expressed cytokines

[0111] From Table 1 and Figure 1 It can be seen that the differentially expressed cytokines in the nasal secretions of patients in the treatment improvement group and the poor treatment group, whose NPS scores improved by more than 2 points, were Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE.

[0112] Example 4: Prediction performance

[0113] The differentially expressed cytokines screened in Example 3 were subjected to receiver operating characteristic curve (ROC) analysis, and the area under the curve (AUC) was calculated. The accuracy of differentially expressed cytokines in predicting the clinical outcomes of patients with chronic sinusitis and nasal polyps treated with omalizumab, as well as their sensitivity and specificity, were evaluated by AUC, Cutoff, and Youden Index. The results are shown in Tables 2 and Figure 2 .

[0114] Table 2. Accuracy, sensitivity, and specificity of biomarkers in nasal secretions for predicting clinical outcomes of improvement in nasal polyps in patients with CRSwNP treated with omalizumab

[0115]

[0116] in conclusion:

[0117] The baseline expression levels of biomarkers Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in nasal secretions can predict the clinical outcome of nasal polyp improvement after omalizumab treatment in patients with CRSwNP;

[0118] Example 5: Verification of prediction performance

[0119] 1. Sample collection

[0120] A total of 18 patients with CRS aged 18 years and above participated in this study.

[0121] See Example 1 for details.

[0122] 2. Evaluation indicators of clinical symptom improvement

[0123] See Example 2.

[0124] 3. Detect the expression levels of seven cytokines, including Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE, in the patient's nasal secretions

[0125] See implementation 3 for the method.

[0126] 4. Verify the prediction performance

[0127] Receiver operating characteristic (ROC) curve analysis was performed to calculate the area under the curve (AUC). The accuracy of differentially expressed cytokines in predicting the clinical outcomes of CRSwNP patients treated with omalizumab, as well as their sensitivity and specificity, were evaluated by AUC, Cutoff, and Youden Index.

[0128] 5. Results and Conclusions

[0129] The baseline expression levels of Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in nasal secretions have good accuracy, sensitivity, and specificity in predicting the clinical outcome of nasal polyp improvement in patients with CRSwNP after omalizumab treatment, and can be used to predict the clinical outcome of patients.

[0130] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. Use of a reagent for detecting the baseline expression level of any one of Eotaxin, GM-CSF, CXCL-1, IP-10, IL-1a, MPO, and IgE in nasal secretions of patients with chronic sinusitis and nasal polyps in the preparation of a product for predicting the clinical outcome of improvement of nasal polyps after omalizumab treatment, characterized in that: The treatment improvement evaluation indicator of the clinical outcome is an improvement in the nasal polyp score (NPS) of more than 2 points.

2. The use according to claim 1, characterized in that Patients who improved with treatment had significantly lower baseline levels of IL-1a in nasal secretions compared with patients who did not respond well to omalizumab.

3. The use according to claim 1, characterized in that Patients who improved with treatment had significantly lower baseline levels of MPO in nasal secretions compared with patients who did not respond well to omalizumab.

4. The use according to claim 1, characterized in that Patients who improved with treatment had significantly lower baseline levels of IgE in nasal secretions compared with patients who did not respond well to omalizumab.

5. The use according to claim 1, characterized in that Patients who improved with omalizumab had significantly lower baseline levels of Eotaxin in nasal secretions compared with patients who did not respond well to omalizumab.

6. The use according to claim 1, characterized in that Patients who improved with treatment had significantly lower baseline levels of GM-CSF in nasal secretions compared with patients who did not respond well to omalizumab.

7. The use according to claim 1, characterized in that Patients who improved with treatment had significantly lower baseline expression of CXCL-1 in nasal secretions compared with patients who did not respond well to omalizumab.

8. The use according to claim 1, characterized in that Patients who improved with treatment had significantly lower baseline levels of IP-10 expression in nasal secretions compared with patients who did not respond well to omalizumab.

9. The use according to claim 1, characterized in that The products include test kits, chips, and test strips.

10. The use according to claim 1, characterized in that Any one of the Eotaxin, GM-CSF, CXCL-1, IP-10, and IL-1a is detected by Luminex; the MPO is detected by enzyme-linked immunosorbent assay Elisa; and the IgE is detected by ImmunoCAP Phadiatop.

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