Use of scn11a protein detection reagent in preparation of kit for screening of depression and evaluating efficacy of antidepressants

Abstract

The present application relates to the field of biological science, and particularly relates to the use of a SCN11A protein detection reagent in the preparation of a depression screening and depression treatment effect evaluation kit. It is found for the first time that the expression level of SCN11A protein in the peripheral blood serum of a depression patient is significantly higher than that of a healthy subject. The reagent for detecting SCN11A protein is used in the preparation of a depression screening kit, so that effective screening of depression and evaluation of the treatment effect of depression can be realized.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of biological science, and particularly relates to a use of a SCN11A protein detection reagent in preparation of a kit for screening of depression and evaluation of efficacy of an antidepressant. BACKGROUND

[0002] Major depressive disorder (MDD) is a kind of mental illness with persistent spontaneous low mood and somatic symptoms. According to statistics, the number of patients with depression in China is as high as 90 million, and is increasing year by year, which seriously affects social production and brings tens of billions of social and economic burden. Depression seriously reduces the quality of life of patients and their families, and some patients even commit self-injury or even suicide, which brings heavy burden to the patient's family and society.

[0003] Although depression has always been concerned, the cause of depression is complex, and there is still a lack of specific diagnostic tools. At present, the diagnosis of depression in the clinic mainly depends on the subjective description and psychological analysis scale of the patient, which is affected by the patient's cultural level and subjective consciousness, and is easy to cause deviation in the diagnosis of the doctor. At the same time, the current clinical treatment effect of depression is less than 40%, and the effective evaluation of the treatment effect of depression is a great challenge in the treatment of depression. Therefore, it is of great value to actively seek effective diagnostic markers for depression.

[0004] SCN11A protein (Uniprot number: Q9UI33) is a protein encoded by SCN11A gene, which is currently mainly used as a drug target for treating human peripheral nerve paroxysmal pain.

[0005] In addition, due to the particularity of mental illness, pathological diagnosis cannot be performed by brain tissue activity, and peripheral blood has the advantages of easy acquisition, small trauma, safety, low price, easy detection and the like. Therefore, it is urgent to find a diagnostic marker for depression from peripheral blood, reduce the misdiagnosis rate of depression, improve the objectivity and accuracy of depression diagnosis, and accurately evaluate the treatment effect of depression.

[0006] Currently, there is no prior art that relates SCN11A protein to depression. SUMMARY

[0007] The purpose of the present application is to provide a use of a SCN11A protein detection reagent in preparation of a kit for screening of depression and evaluation of efficacy of an antidepressant.

[0008] The present application provides a use of a reagent for detecting expression level of SCN11A protein in preparation of a kit for screening of depression, screening of severity of depression and / or evaluation of efficacy of a drug for depression.

[0009] The reagents used to detect the protein expression level of SCN11A include reagents for enzyme-linked immunosorbent assay (ELISA), reagents for immunoblotting, reagents for immunoelectrophoresis, reagents for tissue immunostaining, reagents for immunoprecipitation analysis, reagents for radioimmunoassay, reagents for radioimmunodiffusion, reagents for complement fixation analysis, reagents for fluorescence-activated cell differentiation, reagents for quality analysis, or reagents for protein microarrays.

[0010] The reagents for detecting the protein expression level of SCN11A include antibodies, antibody fragments, high-affinity polymers, peptides, peptide mimics, and nucleic acid aptamers that specifically bind to the full-length or fragments of the SCN11A protein.

[0011] The reagent for detecting SCN11A protein is used to detect SCN11A protein in blood, serum, or plasma.

[0012] The aforementioned antidepressants are sertraline, escitalopram, fluoxetine, paroxetine, venlafaxine, duloxetine, or mirtazapine.

[0013] The present invention also provides a kit for screening depression, screening the severity of depression, and / or evaluating the efficacy of antidepressant drugs, characterized in that it includes reagents for detecting SCN11A protein.

[0014] The reagents used to detect the protein expression level of SCN11A include reagents for enzyme-linked immunosorbent assay (ELISA), reagents for immunoblotting, reagents for immunoelectrophoresis, reagents for tissue immunostaining, reagents for immunoprecipitation analysis, reagents for radioimmunoassay, reagents for radioimmunodiffusion, reagents for complement fixation analysis, reagents for fluorescence-activated cell differentiation, reagents for quality analysis, or reagents for protein microarrays.

[0015] The reagents for detecting the protein expression level of SCN11A include antibodies, antibody fragments, high-affinity polymers, peptides, peptide mimics, and nucleic acid aptamers that specifically bind to the full-length or fragments of the SCN11A protein.

[0016] The reagent for detecting SCN11A protein is used to detect SCN11A protein in blood, serum, or plasma; and / or, the antidepressant is sertraline, escitalopram, fluoxetine, paroxetine, venlafaxine, duloxetine, or mirtazapine.

[0017] This invention also provides a depression prediction model, characterized in that: it is Y = 1 / (1+e (-0.206*SCN11A+40.027) Where Y represents the probability of having depression, and SCN11A represents the level of SCN11A measured in peripheral serum, with the unit of SCN11A level being pg / mL; if the Y value is greater than 0.5, depression can be diagnosed, and if the Y value is less than 0.5, depression is not diagnosed.

[0018] Experimental results demonstrate the relationship between SCN11A and depression. This invention collected peripheral blood samples from healthy controls, first-episode untreated depression patients, and patients treated with antidepressants. ELISA analysis of SCN11A levels in each group revealed significantly elevated SCN11A levels in serum compared to healthy controls. However, serum SCN11A levels in patients treated with antidepressants were significantly lower than in first-episode untreated depression patients, suggesting that serum SCN11A levels may be a peripheral biomarker for depression. Furthermore, this invention further demonstrated the reliability and specificity of SCN11A as a biomarker for diagnosing depression and / or assessing the risk of developing depression by constructing a depression diagnostic model and performing ROC curve analysis.

[0019] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0020] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0021] Figure 1 This image shows the quantitative results of SCN11A in the peripheral serum of patients with depression and healthy controls.

[0022] Figure 2 This graph shows the quantitative results of SCN11A in peripheral serum of patients with first-episode untreated depression and patients with depression treated with antidepressants. (DN-MDD, first-episode untreated depression; DT-MDD, depression treated with antidepressants).

[0023] Figure 3 The ROC curves of the depression diagnostic model constructed using SCN11A levels in the training set are shown in the training and test sets. Detailed Implementation

[0024] The raw materials and equipment used in this invention are all known products, obtained by purchasing commercially available products.

[0025] Example 1: The relationship between SCN11A protein in peripheral serum and depression

[0026] I. Clinical Treatment

[0027] The study included peripheral serum samples collected from 73 first-episode untreated patients with depression who had never taken any antipsychotic medication, 66 patients with depression who had been treated with antidepressants (using clinically proven positive antidepressants, including sertraline, escitalopram, fluoxetine, paroxetine, venlafaxine, duloxetine, and mirtazapine), and 65 healthy controls. The diagnosis of depression was made by two experienced psychiatrists according to the diagnostic criteria in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). All patients were recruited from the Department of Psychiatry, First Affiliated Hospital of Chongqing Medical University, and samples from healthy controls were collected from the Physical Examination Center of the First Affiliated Hospital of Chongqing Medical University. The demographic and clinical characteristics of the participants are shown in Table 1.

[0028] Table 1: Demographic and clinical characteristics of the subjects

[0029]

[0030]

[0031] Note:

[0032] HCs: Healthy individuals without mental illness; MDD: Individuals with depression;

[0033] mark "a” Indicates the chi-square test;

[0034] mark "b” This represents the Mann-Whitney U test.

[0035] Collect 5 ml of blood from the median cubital vein of all subjects, let stand at room temperature for 30 min, centrifuge at 1000×g for 15 min, collect the serum, aliquot and freeze at -80℃.

[0036] II. Detection Methods

[0037] The severity of depression was assessed using the 17-item Hamilton Depression Rating Scale (HAMD-17).

[0038] The human SCN11A ELISA kit (catalog number: MM-60326H1) from Jiangsu Enzyme Immunosorbent Assay Co., Ltd. was used to detect SCN11A levels in serum samples according to the kit instructions. The detection range of this kit is 4 pg / mL to 220 pg / mL.

[0039] III. Model Establishment of this Invention

[0040] (I) Data Analysis

[0041] SPSS 20.0 software was used to perform statistical analysis on all data. Data are expressed as mean ± standard error. Statistical analysis was performed using t-tests, chi-square tests, or nonparametric Mann-Whitney U tests. A p-value < 0.05 was considered statistically significant.

[0042] (II) Constructing a Diagnostic Model for Depression

[0043] The random forest machine learning model is employed, which is not only simple and easy to operate, but also generates a depression diagnostic model with high sensitivity and specificity. Specifically, the study subjects were divided into a training set (healthy controls, n=44; patients with depression, n=92) and a test set (healthy controls, n=21; patients with depression, n=47). After adjusting for age, BMI, and gender using the SCN11A level in the training set samples, a diagnostic model was established: Y = 1 / (1+e^(-1 / 2)). (-0.206*SCN11A+40.027) (Note: Y represents the probability of developing depression. If the Y value is greater than 0.5, depression can be diagnosed; if the Y value is less than 0.5, depression is not diagnosed. SCN11A represents the level of SCN11A measured in peripheral serum.) The unit of SCN11A level is pg / mL, and it is tested and verified in the test set sample. The receiver operating characteristic of the model was analyzed using GraphPad Prism 8.3 software, and ROC curves were plotted. The area under the ROC curve (AUC) is known to be: 1-0.9, 0.9-0.8, 0.8-0.7, 0.7-0.6, and 0.6-0.5, which represent excellent, good, average, poor, and unsatisfactory, respectively.

[0044] IV. Results

[0045] The results of serum SCN11A levels in patients with depression and healthy controls are shown in the table below. Figure 1 The results of serum SCN11A levels in patients with first-episode untreated depression and patients with depression treated with antidepressants are shown in Table 2. Figure 2 And Table 3, the diagnostic efficacy analysis of serum SCN11A levels is shown in Table 3. Figure 3 .

[0046] Table 2. Serum SCN11A levels in patients with depression and healthy controls.

[0047] HCs MDD p-value Sample size (n) 65 139 - Mean SCN11A level ± standard error (pg / mL) 180.62±1.30 211.88±1.33 3.83E-25

[0048] Note: HCs: Healthy individuals without mental illness; MDD: Individuals with depression;

[0049] from Figure 1 As shown in Table 2, the serum SCN11A protein level in patients with depression was significantly upregulated compared with that in healthy controls without mental illness, and the difference between the two groups was highly significant (p = 3.83E-25).

[0050] Table 3. Serum SCN11A levels in patients with first-episode untreated depression and patients with depression treated with antidepressants.

[0051] DN-MDD DT-MDD p-value Sample size (n) 73 66 - Mean ± standard error (pg / mL) 223.14±1.29 199.44±1.13 1.08E-27

[0052] Note: DN-MDD refers to first-episode untreated depression; DT-MDD refers to depression treated with antidepressants.

[0053] from Figure 2 As shown in Table 3, the serum SCN11A level in patients treated with antidepressants was significantly lower than that in first-episode untreated patients (p = 1.08E-27). Therefore, the efficacy of antidepressant treatment can be evaluated by detecting the serum SCN11A protein level. If the SCN11A protein level decreases, the treatment is effective; if it does not decrease, the treatment is ineffective.

[0054] Figure 3 The area under the ROC curve for this model was 0.9571 in the training set and 0.9357 in the test set, falling between 1 and 0.9, indicating good subject operating characteristic results when the model was constructed using SCN11A levels from the training set. This confirms that SCN11A can serve as a biomarker for depression, providing a valid basis for relevant treatment measures or decision-making for patients.

[0055] Experimental results show that SCN11A protein can be used as an auxiliary diagnostic tool for clinical depression and can be used to evaluate the therapeutic effects of antidepressants. Furthermore, through model establishment, it was confirmed that SCN11A can serve as a biomarker for depression, providing a valid basis for patients to take relevant treatment measures or make relevant decisions.

[0056] In summary, the kit of this invention can screen the risk of depression in a tested population by detecting the level of SCN11A protein in peripheral serum: a high level of SCN11A protein indicates a high risk of depression, while a low level indicates a low risk. After patients use antidepressants, the SCN11A protein level decreases accordingly, and the SCN11A protein level can be used to evaluate the therapeutic effect of antidepressants. Furthermore, this invention, through the establishment of a model, has confirmed that SCN11A can serve as a diagnostic biomarker for depression, providing an effective basis for patients to take relevant treatment measures or make decisions, and has promising clinical application prospects.

Claims

1. The use of reagents for detecting SCN11A protein expression levels in the preparation of kits for screening depression and / or evaluating the efficacy of antidepressant drugs.

2. The use as described in claim 1, characterized in that, The reagents used to detect the protein expression level of SCN11A include reagents for enzyme-linked immunosorbent assay (ELISA), reagents for immunoblotting, reagents for immunoelectrophoresis, reagents for tissue immunostaining, reagents for immunoprecipitation analysis, reagents for radioimmunoassay, reagents for radioimmunodiffusion, reagents for complement fixation analysis, reagents for fluorescence-activated cell differentiation, reagents for quality analysis, or reagents for protein microarrays.

3. The use as described in claim 1, characterized in that, The reagents for detecting the protein expression level of SCN11A include antibodies, antibody fragments, high-affinity polymers, peptides, peptide mimics, and nucleic acid aptamers that specifically bind to the full-length or fragments of the SCN11A protein.

4. The use as described in claim 1, characterized in that, The reagent for detecting SCN11A protein is used to detect SCN11A protein in blood, serum, or plasma.

5. The use as described in claim 1, characterized in that, The aforementioned antidepressants are sertraline, escitalopram, fluoxetine, paroxetine, venlafaxine, duloxetine, or mirtazapine.

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