Method and use for predicting or evaluating efficacy of omalizumab in treatment of recurrent nasal polyps by using topical ige

Abstract

Disclosed is a biomarker for predicting or evaluating the efficacy of omalizumab in the treatment of recurrent nasal polyps. The biomarker is an index of topical IgE, which index is optionally detected using an ImmunoCAP 100 system, and the topical IgE is standardized by means of evaluating using a BCA assay the total protein concentration in nasal secretion. The biomarker can rapidly and accurately predict or evaluate the efficacy of omalizumab in the treatment of recurrent nasal polyps.

Description

A method and application for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps using local IgE Technical Field

[0001] The present invention relates to the field of disease analysis technology, and in particular to a method and application of using local IgE to predict or evaluate the efficacy of omalizumab in treating recurrent nasal polyps. Background Art

[0002] Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by immunoglobulin E (IgE) hypersecretion and eosinophilia and affects 4.3% of the world's population. Omalizumab (Xolair) is a humanized recombinant monoclonal antibody that blocks free IgE receptor binding and reduces expression of high-affinity IgE Fc receptors on effector cells, thereby attenuating type 2 inflammation. Previous studies have demonstrated the efficacy of omalizumab in the treatment of CRSwNP. However, the correlation between serum IgE and local IgE is poor. Although previous studies have reported elevated local IgE levels in CRSwNP, the correlation between local IgE levels and omalizumab efficacy remains largely unknown.

[0003] Therefore, to fill the above gaps, this study aimed to explore the correlation between local IgE levels and the efficacy of omalizumab. Summary of the Invention

[0004] The present invention first provides a biomarker for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps. The marker is an indicator of local IgE, which is optionally obtained by detection using the ImmunoCAP 100 system and the total protein concentration in nasal secretions is assessed by the BCA assay to standardize the local IgE.

[0005] Preferably, the local IgE sample acquisition method comprises the following steps: (1) using an expandable sponge to collect nasal secretions, placing it into both nasal cavities for 5 minutes, and then soaking the sponge in 1 mL of 0.9% saline at 4°C for 2 hours; (2) then centrifuging the sponge using a centrifuge (4°C, 1500g, 15 minutes), and storing the centrifuged liquid in a -80°C ultra-low temperature freezer.

[0006] The present invention also provides an application of a marker, the application being a) preparing a detection reagent for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps; or b) preparing or evaluating a detection kit for predicting the efficacy of omalizumab in treating recurrent nasal polyps; or c) a method for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, the method optionally being used for non-diagnostic or non-therapeutic purposes.

[0007] The present invention also provides a kit for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, wherein the kit comprises a detection reagent for detecting a marker.

[0008] The present invention also provides a use of the kit as claimed in claim 4 in the preparation of a method for predicting or evaluating the efficacy of anti-omalizumab in treating recurrent nasal polyps, wherein the use is optionally used for non-diagnostic purposes or non-therapeutic purposes.

[0009] The present invention also provides use of a marker detection reagent in preparing a kit for predicting or evaluating the efficacy of anti-omalizumab in treating recurrent nasal polyps.

[0010] The present invention also provides a method for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, which is optionally used for non-diagnostic or non-therapeutic purposes, and comprises the steps of detecting local IgE, and predicting or evaluating the efficacy of omalizumab based on the index results of local IgE.

[0011] Preferably, the treatment effect of the patients with higher local IgE is better than that of the patients with lower local IgE.

[0012] Preferably, the lower standard for patients with lower local IgE is IgE<5.11kIU / g, and the standard for patients with higher local IgE is IgE≥5.11kIU / g.

[0013] Compared with the prior art, the present invention has at least the following beneficial effects:

[0014] The present invention discovered that the local IgE index can be used as a marker to predict or evaluate the efficacy of omalizumab in treating recurrent nasal polyps. The marker of the present invention can quickly and accurately predict or evaluate the efficacy of omalizumab in treating recurrent nasal polyps, filling the gap in the existing technology and having significant clinical and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Experimental design process;

[0016] Figure 2 Changes in subjective symptom scores and objective examinations compared with baseline at different treatment time points;

[0017] Figure 3 Correlation between local IgE and peripheral blood IgE and NPS improvement after four months of treatment;

[0018] Figure 4 Changes in subjective symptom scores and objective examinations at different treatment time points in the local high IgE and low IgE groups compared with baseline;

[0019] Figure 5 Comparison of response rates between the local high IgE and low IgE groups at different treatment time points;

[0020] Figure 6 IgE in nasal polyps at baseline in responders and non-responders. Number of IgE+ cells (n = 29). HPF, high-power field. *: P < 0.05;

[0021] Figure 7. Receiver operating characteristic (ROC) curve for predicting the efficacy of omalizumab treatment using IgE+ cell counts in nasal polyp tissue. AUC, area under the curve; CI, confidence interval.

[0022] Figure 8. Eosinophils and IgE in nasal polyps of responders and nonresponders before and after omalizumab treatment. AC, IgE+ cells in nasal polyps of responders and nonresponders before and after omalizumab treatment (n = 23). EOT, end of treatment; LPF, low-power field. **: P < 0.01, ns: not statistically significant. DETAILED DESCRIPTION

[0023] In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, a detailed description will be given below with reference to the accompanying drawings and specific embodiments.

[0024] Example 1 Patient data and study design

[0025] This study was a single-center trial conducted at Beijing Tongren Hospital from May 2019 to April 2021. A total of 35 patients aged 18 years and older with recurrent CRSwNP were enrolled. Recurrence was defined as the persistence of polyps, mucopurulent rhinorrhea, and severe nasal symptoms after at least one month of maximal medical therapy after surgery. Patients were required to have completed at least four weeks of nasal steroid therapy before enrollment. At the end of nasal steroid therapy, eligible patients had a bilateral nasal polyp score (NPS) ≥ 2 (NPS ≥ 1 per side). Patients with CRSwNP who met the enrollment criteria and were able to complete the required four months of nasal steroid therapy during the same period were selected as the control group. Patients with sinus mucoceles, unilateral sinus lesions, choanal polyps, allergic fungal sinusitis, immunologic diseases, and those who had received systemic corticosteroids within four weeks prior to enrollment were excluded. Furthermore, patients who received escalating therapy, such as oral corticosteroids or subcutaneous biologics, during the four-month treatment period were excluded from the control group. This study was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (TRECKY2021-030). Written informed consent was obtained from the patients before participation.

[0026] Thirty-five patients were divided into the omalizumab treatment group (n=29) and the control group (n=6). Patients in the control group received only nasal steroids (budesonide nasal spray [64 μg / spray, 128 μg / side, QD]). The basic characteristics of the patients are shown in Table 1. Patients in the omalizumab treatment group received nasal steroids and omalizumab for 4 months. The dose of omalizumab was calculated based on the patient's total serum IgE and body weight, with a subcutaneous injection of 150 mg to 600 mg once a month (a total of 4 times) or once every two weeks (a total of 8 times). The experimental process is shown in Figure 1. During the study, nasal irrigation treatment was allowed for all patients. Prohibited drugs include: systemic corticosteroids, montelukast sodium tablets, and systemic antibiotic treatment for more than 14 days.

[0027] Table 1. Basic characteristics of the patients Note: *: P<0.05.

[0028] Example 2 Clinical Evaluation Indicators

[0029] Demographic data were collected from all patients. Nasal symptom severity scores (NPS), visual analog scale (VAS) for nasal symptoms (nasal obstruction, runny nose, loss of smell, headache), and the 22-item Sino-Nasal Outcome Test (SNOT-22) were collected at baseline and at months 1, 2, 3, and 4 for further analysis. The NPS score ranges from 0 to 8, with 0 = no polyp, 1 = a small polyp visible in the middle meatus that does not extend beyond the inferior margin of the middle turbinate, 2 = a polyp reaching the inferior margin of the middle turbinate, 3 = a polyp reaching the inferior margin of the inferior turbinate, or a polyp score of 2 accompanied by a large polyp medial to the middle turbinate, and 4 = a polyp that completely or nearly completely obstructs the nasal cavity, i.e., to the nasal floor. The VAS score is based on a 10-point scale, with 0 indicating no symptoms and 10 indicating very severe symptoms. The minimal clinical difference value for the SNOT-22 was 8.9 points.

[0030] Example 3 Objective inspection and sample testing

[0031] Blood and tissue samples were collected at baseline and month 4 for complete blood count and histopathological examination to count eosinophils. Nasal polyp tissue was stained with hematoxylin and eosin and then examined by two pathologists at 400x magnification. Inflammatory cells (eosinophils, neutrophils, plasma cells, and lymphocytes) were counted in three randomly selected fields under a 400x light microscope. The average count was used to calculate the eosinophil percentage.

[0032] Acquisition of local IgE and serum IgE:

[0033] Topical IgE: using a swellable sponge ( Nasal secretions were collected using a sponge (Medtronic Xomed, Jacksonville, FL, USA). After being placed in both nasal cavities for 5 minutes, the sponge was soaked in 1 mL of 0.9% saline at 4°C for 2 hours. The sponge was then centrifuged (4°C, 1500 g, 15 minutes), and the centrifuged fluid was stored in a -80°C freezer.

[0034] Serum IgE: After the patient's blood sample was centrifuged (3000 rpm, room temperature, 10 minutes), the serum was collected for further analysis.

[0035] IgE detection: Local IgE and serum IgE were detected using the ImmunoCAP 100 system (Pharmacia, Uppsala, Sweden). Total protein concentration in nasal secretions was assessed by BCA assay (Shanghai Biotech Biotechnology Co., Ltd., China), and local IgE was normalized.

[0036] Statistical methods:

[0037] IBM SPSS Statistics 26 (IBM Corp.) and Prism 9 (GraphPad Software, Inc.) for macOS were used for data analysis. A mixed-effects model was used to compare the changes in various clinical indicators from baseline at each treatment visit. The Mann–Whitney U test was used to determine whether there were significant differences between the two groups. Paired t-tests were used to compare pre- and post-treatment changes. Fisher's exact test was used to compare ratios between the groups. Spearman's rank correlation coefficient was used for nonparametric correlation analysis. A P value < 0.05 indicated statistical significance.

[0038] Example 4 Sample Test Results

[0039] After 4 months of omalizumab treatment, the NPS (p = 0.000), SNOT-22 (p = 0.0000), olfactory VAS score (p = 0.000), nasal congestion VAS score (p = 0.000), runny nose VAS score (p = 0.000), and headache VAS score (p = 0.007) of patients with CRSwNP were significantly reduced compared with baseline ( Figure 2A-F and Table 2 ).

[0040] Table 2. Changes in clinical parameters compared with baseline after omalizumab treatment

[0041] Statistical differences in the NPS, SNOT-22, sense of smell, nasal congestion, and runny nose scores compared to baseline began to emerge after one month of omalizumab treatment (Figures 2A-E, p = 0.000). After three months of omalizumab treatment, statistical differences in the headache VAS score compared to baseline began to emerge (Figure 2F, p = 0.006 at month 3 and p = 0.007 at month 4). Furthermore, omalizumab treatment significantly reduced peripheral blood eosinophil counts (Figure 2G, median ± interquartile range: 0.38 ± 0.31 vs. 0.49 ± 0.36 × 109 cells / L after treatment, p < 0.001). No significant changes in peripheral blood eosinophil counts were observed in the control group.

[0042] We analyzed the correlation between baseline serum IgE and local IgE levels and the degree of NPS improvement after treatment in patients receiving omalizumab. The study found that local IgE levels were negatively correlated with the degree of improvement in NPS (post-treatment minus pre-treatment) (p = 0.042, r = -0.380), while total serum IgE levels were not correlated with the degree of NPS improvement (p = 0.491). This suggests that patients with higher local IgE levels had more significant polyp reduction after omalizumab treatment (Figure 3A-B). Therefore, we divided the omalizumab-treated group into two subgroups based on the median local IgE level (5.11 kIU / g): patients in the low IgE group had local IgE <5.11 kIU / g, and patients in the high IgE group had local IgE ≥5.11 kIU / g. The baseline characteristics of the patients in the two subgroups are shown in Table 3.

[0043] Table 3. Basic clinical characteristics of patients in the local high IgE group and low IgE group

[0044] After 4 months of omalizumab treatment, the NPS (p = 0.000 for both groups), SNOT-22 (p = 0.000 for both groups), VAS scores for olfaction (p = 0.001 for both groups), VAS scores for nasal congestion (p = 0.000 for both groups), and VAS scores for rhinorrhoea (high local IgE group: p = 0.000; low local IgE groups: p = 0.008) were significantly reduced compared with baseline (Figures 4A–E and Table 4). A reduction in the VAS score for headache was observed only in patients in the high local IgE group (Figure 4F and Table 4, p = 0.012).

[0045] Table 4. Changes of clinical parameters compared with baseline in the local high IgE group and low IgE group after omalizumab treatment

[0046] Notably, nasal congestion and runny nose improved in the first month of omalizumab treatment in patients in the local high IgE group, whereas improvement was not seen until the second month in patients in the local low IgE group. Furthermore, after four months of omalizumab treatment, the blood eosinophil count in patients in the local high IgE group was significantly reduced compared with baseline (Figure 4G, median ± interquartile range: 0.36 ± 0.26 vs. 0.53 ± 0.24 × 109 cells / L, p = 0.001), whereas the blood eosinophil count in patients in the local low IgE group remained unchanged compared with baseline (Figure 4G: 0.39 ± 0.41 vs. 0.42 ± 0.37 × 109 cells / L, p = 0.777). After 4 months of omalizumab treatment, tissue eosinophil counts in both groups were significantly lower than those at baseline, but the downward trend was more pronounced in the local high IgE group than in the local low IgE group (mean ± SD, −187 ± 209 cells / [high power field, HPF] vs. −118 ± 188 cells / HPF, Figure 4H).

[0047] We further evaluated the response rates of patients with an improvement of ≥2 points in NPS, ≥8.9 points in SNOT-22, and ≥4 points in nasal VAS at different time points. We found that the proportion of patients with an improvement of ≥2 points in NPS after one month of omalizumab treatment was significantly higher in the local high IgE group than in the control group (Figure 5A and Table 5, month 1: p = 0.006; month 2: p = 0.002; month 3: p = 0.010; month 4: p > 0.001), while similar results were found in the local low IgE group only at month 2 of omalizumab treatment (Figure 5A, local low IgE group vs. control group, month 2: 28.57% vs. 0.00%, p = 0.018). The proportion of patients with an improvement of olfactory VAS score ≥4 in the local high IgE group was significantly higher than that in the control group ( Figure 5B and Table 5 , month 1: p = 0.002; month 2: p = 0.000; month 3: p = 0.000; month 4: p = 0.029).

[0048] Table 5. Comparison of response rates between the local high IgE group and the control group

[0049] However, similar results were not found in the local low IgE group. The proportion of patients with an improvement of ≥4 in the olfactory VAS score at month 1 and month 2 was significantly higher in the local high IgE group than in the local low IgE group (Figure 5B, high local IgE group vs. low local IgE group, month 1: 40.00% vs. 7.14%, p = 0.023; month 2: 60.00% vs. 14.29%, p = 0.004). The proportion of patients with an improvement of ≥4 in the nasal congestion VAS score after 3 months of omalizumab treatment was significantly higher in the local high IgE group than in the control group (Figure 5C and Table 5, month 3: p = 0.029; month 4: p = 0.010), while no difference was found between the low IgE and control groups. The proportion of patients with an improvement of ≥4 in the runny nose VAS score at month 4 of omalizumab treatment was significantly higher in the local high IgE group than in the control group (Figure 5D and Table 5, p = 0.029), while no difference was found between the low IgE and control groups. The proportion of patients with an improvement of ≥4 points in the VAS score for runny nose was significantly higher in the high-IgE group than in the low-IgE group at one month of omalizumab treatment (Figure 5D, 53.33% vs. 14.29% in the high-IgE group and the low-IgE group, p = 0.014). Regarding the SNOT-22, the proportion of patients achieving a score of ≥8.9 in both groups was significantly higher than that in the control group during the first month of omalizumab treatment (Figure 5E, 66.67% vs. 33.33% in the high-IgE group and the control group, p = 0.029; 64.29% vs. 33.33% in the low-IgE group and the control group, p = 0.005). There was no difference in the proportion of patients with an improvement of ≥4 points in the VAS score for headache among the three groups (Figure 5F and Table 5).

[0050] All 29 patients with CRSwNP who completed 16 weeks of omalizumab treatment (Cohort A) were divided into two groups: patients with a change in nasal polyp score greater than 1 point were considered responders to omalizumab treatment, while those without the change were considered non-responders. As shown in Table 1, no significant differences were observed between responders and non-responders at baseline in terms of gender, age, comorbidities, symptom scores, nasal polyp scores, serum total IgE levels, and peripheral blood and tissue eosinophil levels.

[0051] Table 6 Demographic and clinical characteristics of CRSwNP patients who responded or did not respond to omalizumab treatment IQR: interquartile range; SEM: standard error of the mean; HPF, high-power field of view.

[0052] Immunofluorescence staining revealed that at baseline, the number of IgE+ cells in nasal polyp tissues of responders was significantly higher than that in nasal polyp tissues of non-responders ( FIG. 6 , P=0.024).

[0053] We then performed a logistic regression analysis to investigate whether the IgE+ cell count in nasal polyp tissue could serve as a predictor of omalizumab treatment efficacy. Univariate analysis showed that the number of IgE+ cells in nasal polyp tissue at low magnification was significantly associated with omalizumab treatment outcome (P = 0.028, odds ratio (OR) = 1.486), whereas changes in the number of IgE+ cells at high magnification were not associated with outcome. The predictive value of the IgE+ cell count in nasal polyp tissue at low magnification was assessed using a receiver operating characteristic (ROC) curve, with an area under the curve of 0.811 (P = 0.007; Figure 7).

[0054] At the end of treatment, the numbers of IgE+ cells and EPX+ cells in nasal polyp tissues of responders were significantly reduced compared with baseline, while there was no significant difference in non-responders ( Figure 8 ).

[0055] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A biomarker for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, characterized in that: The marker is an index of local IgE, which is optionally detected using the ImmunoCAP 100 system and normalized to local IgE by assessing total protein concentration in nasal secretions by the BCA assay.

2. The marker according to claim 1, characterized in that The local IgE sample acquisition method comprises the following steps: (1) using a swellable sponge to collect nasal secretions, placing it in two nasal cavities for 5 minutes, and then soaking the sponge in 1 mL of 0.9% saline at 4°C for 2 hours; (2) then using a centrifuge to centrifuge the sponge (4°C, 1500g, 15 minutes), and storing the centrifuged liquid in a -80°C ultra-low temperature refrigerator.

3. A use of the marker according to claim 1 or 2, characterized in that: The application is a) preparing a detection reagent for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps; or b) preparing or evaluating a detection kit for predicting the efficacy of omalizumab in treating recurrent nasal polyps; or c) a method for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, which is optionally used for non-diagnostic or non-therapeutic purposes.

4. A kit for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, characterized in that: The kit comprises a detection reagent for detecting the marker according to claim 1 or 2.

5. Use of the kit according to claim 4 in the preparation of a method for predicting or evaluating the efficacy of anti-omalizumab in the treatment of recurrent nasal polyps, wherein the use is optionally used for non-diagnostic purposes or non-therapeutic purposes.

6. Use of a detection reagent for the marker as claimed in claim 1 or 2 in the preparation of a kit for predicting or evaluating the efficacy of anti-omalizumab in the treatment of recurrent nasal polyps.

7. A method for predicting or evaluating the efficacy of omalizumab in treating recurrent nasal polyps, characterized in that: The method is optionally used for non-diagnostic purposes or non-therapeutic purposes, and comprises the step of detecting local IgE, and predicting or evaluating the efficacy of omalizumab based on the index result of local IgE.

8. The method according to claim 5, characterized in that The patients with higher local IgE have better treatment effects than those with lower local IgE.

9. The method according to claim 6, characterized in that The lower standard for patients with lower local IgE is IgE<5.11kIU / g, and the standard for patients with higher local IgE is IgE≥5.11kIU / g.

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