A biomarker for predicting recurrence of chronic rhinosinusitis and use thereof

Abstract

The application discloses a biomarker for predicting recurrence of chronic rhinosinusitis and application thereof, and the biomarker is CASP3 and / or CHMP7. The biomarker CASP3 and CHMP7 gene has high transcription expression in recurrent chronic rhinosinusitis, is a potential recurrence diagnosis marker, is used for diagnosis and prediction of recurrence states of all types of chronic rhinosinusitis, more accurately judges whether rhinosinusitis recurs or not, effectively reduces misdiagnosis rate and missed diagnosis rate, and provides a new perspective and tool for early diagnosis of diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical diagnosis, and in particular to a biomarker for predicting recurrence of chronic rhinosinusitis and application thereof. BACKGROUND

[0002] Chronic rhinosinusitis is a common disease in the department of ear-nose-throat and neck surgery, and the recurrence rate of the type mainly with type 2 inflammation is high, which seriously affects the quality of life of patients. At present, the prediction of recurrence of chronic rhinosinusitis in the clinic mainly depends on the detection of cytokines related to type 2 inflammation in nasal mucosa tissue, and the proportion of eosinophils in peripheral blood and nasal mucosa tissue of patients. Although these indicators have a positive guiding effect on predicting the recurrence of the disease, the type 2 inflammation-based chronic rhinosinusitis also has a risk of recurrence, at which time the effect of using type 2 inflammation-related indicators to diagnose and predict the recurrence of the disease is limited. Therefore, it is necessary to find new markers suitable for diagnosing or predicting the recurrence of type 2 and non-type 2 chronic rhinosinusitis, and then control the disease in the best range through active intervention.

[0003] In recent years, with the in-depth study of the mechanism of inflammation, the role of pyroptosis-related genes in inflammatory diseases has gradually attracted attention. Pyroptosis is a form of programmed cell death closely related to inflammatory response, and its related genes show specific changes in various inflammatory diseases. There are relevant reports that cell pyroptosis can cause hormone resistance, thereby reducing the effect of glucocorticoid treatment on chronic rhinosinusitis. However, whether cell pyroptosis genes can be used as markers to diagnose or predict the recurrence of chronic rhinosinusitis is still unclear. In view of the fact that the application of cell pyroptosis genes in diagnosing or predicting the recurrence of chronic rhinosinusitis is still in the academic research stage, the person skilled in the art further studies the cell pyroptosis-related markers based on the previous research, so as to effectively predict the possibility of recurrence of all types of chronic rhinosinusitis, remind patients of the risk of recurrence, attract the attention of patients, and take preventive measures in advance to minimize the risk of disease recurrence. SUMMARY

[0004] In order to overcome the shortcomings of the prior art, one of the purposes of the present application is to provide a biomarker for predicting recurrence of chronic rhinosinusitis and application thereof, so as to solve the above-mentioned traditional technical problems, effectively predict the possibility of recurrence of all types of chronic rhinosinusitis, remind patients of the risk of recurrence, attract the attention of patients, and take preventive measures in advance to minimize the risk of disease recurrence.

[0005] The second purpose of the present application is to provide an application for detecting the expression amount of the above-mentioned biomarker in the preparation of a kit and / or device for predicting recurrence of chronic rhinosinusitis.

[0006] One of the purposes of the present application is achieved by adopting the following technical solutions:

[0007] A biomarker for predicting recurrence of chronic rhinosinusitis, the biomarker being CASP3 and / or CHMP7.

[0008] In the present application, it is found that CASP3 and CHMP7 genes have high transcription expression in recurrent chronic rhinosinusitis, and are potential recurrence diagnostic markers, which are used for the diagnosis and prediction of recurrence of all types of chronic rhinosinusitis, more accurately judge whether the rhinosinusitis is recurrent, effectively reduce the misdiagnosis rate and missed diagnosis rate, and provide a new perspective and tool for early diagnosis of the disease.

[0009] The second purpose of the present application is achieved by the following technical scheme:

[0010] An application will be used for the reagent for detecting the expression amount of the biomarker to prepare a kit and / or device for predicting recurrence of chronic rhinosinusitis.

[0011] Further, the reagent includes a reagent for quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR).

[0012] That is, the reagent required for quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) when detecting the transcription expression of CHMP7 and CASP3.

[0013] Compared with the prior art, the present application has the following beneficial effects:

[0014] 1、The biomarker CASP3 and CHMP7 genes in the present application have high transcription expression in recurrent chronic rhinosinusitis, and are potential recurrence diagnostic markers, which are used for the diagnosis and prediction of recurrence of all types of chronic rhinosinusitis, more accurately judge whether the rhinosinusitis is recurrent, effectively reduce the misdiagnosis rate and missed diagnosis rate, and provide a new perspective and tool for early diagnosis of the disease.

[0015] 2、The present application screens pyroptosis-related genes that significantly change in the recurrence group as potential markers by comparing the pyroptosis-related gene expression profiles of the recurrence group and the non-recurrence group of chronic rhinosinusitis patients, and identifies and verifies them by qRT-PCR method. Based on the screened pyroptosis-related markers, a specific and high-sensitivity detection method is established, and early diagnosis of rhinosinusitis recurrence is realized by quantitative detection of pyroptosis-related genes in the nasal sinus tissue of patients. The present application protects the detection based on specific pyroptosis-related markers, and protects the application in clinical rhinosinusitis recurrence monitoring, treatment effect evaluation, prognosis judgment and drug research and development.

[0016] 3、Current clinical evaluation of chronic rhinosinusitis recurrence is mainly based on the detection results of type 2 inflammatory factors and eosinophils. These indicators are suitable for evaluating diseases mainly with type 2 inflammation, and the detection sensitivity for patients with non-type 2 inflammation is low, and it is difficult to find signs of recurrence in time when the disease has not shown obvious clinical symptoms. The present application detects and screens specific changed genes as markers by studying and detecting pyroptosis related gene expression in the recurrence group and non-recurrence group of chronic rhinosinusitis patients. Traditional methods are often limited by subjective factors and detection techniques themselves, and are prone to misdiagnosis and missed diagnosis. The present application detects specific markers to more accurately determine whether rhinosinusitis has recurred, effectively reduces the misdiagnosis rate and the missed diagnosis rate, and provides a new perspective and tool for early diagnosis of the disease. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 Figure 1 is a graph of the high expression of pyroptosis related genes in chronic rhinosinusitis, wherein, Figure 1 A is a visualization result graph of the expression of cell pyroptosis related genes, Figure 1 B is a result graph of pathway enrichment analysis of cell pyroptosis related genes, Figure 1 C is a result graph of pathway enrichment analysis of cell pyroptosis related genes, Figure 1 D is a result graph of pathway enrichment analysis of cell pyroptosis related genes, Figure 1 E is a result graph of pathway enrichment analysis of cell pyroptosis related genes, Figure 1 F is a result graph of expression analysis of cell pyroptosis related genes, Figure 1 G is a result graph of pathway enrichment analysis of cell pyroptosis related differential genes, Figure 1 H is a result graph of pathway enrichment analysis of cell pyroptosis related differential genes;

[0018] Figure 2 Figure 2 is an expression and recurrence prediction verification graph of pyroptosis related genes in recurrent chronic rhinosinusitis, wherein, Figure 2 A is an expression analysis graph of AIM2 in the RCRSwNP group and the non-RCRSwNP group, Figure 2 B is an expression analysis graph of BAK1 in the RCRSwNP group and the non-RCRSwNP group, Figure 2 C is an expression analysis graph of GZMA in the RCRSwNP group and the non-RCRSwNP group, Figure 2 D is an expression analysis graph of GSDMC in the RCRSwNP group and the non-RCRSwNP group, Figure 2 E is an expression analysis graph of GSDMC in the GSDME group and the non-RCRSwNP group, Figure 2 F is an expression analysis graph of PYCARD in the GSDME group and the non-RCRSwNP group, Figure 2G represents the expression analysis of TNF in the GSDME group and the non-RCRSwNP group. Figure 2 H represents the expression analysis of CHMP4C in the GSDME group and the non-RCRSwNP group. Figure 2 Figure I shows the expression analysis of CASP3 in the GSDME group and the non-RCRSwNP group. Figure 2 J is the expression analysis diagram of the GSDME group and the non-RCRSwNP group of CHMP7. Figure 2 K represents the ROC curve of CASP3. Figure 2 L represents the ROC curve of CHMP7. Detailed Implementation

[0019] The present invention will now be further described in conjunction with specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments.

[0020] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. Informed consent was obtained from all subjects in the collection of the tissue samples.

[0021] Example 1: Screening Biomarkers

[0022] A biomarker for predicting recurrence of chronic sinusitis, the screening and identification process of which includes the following steps:

[0023] Bioinformatics analysis was used to preliminarily screen differentially expressed pyroptosis-related genes that are highly expressed in chronic sinusitis. The results showed that a total of 10 pyroptosis-related genes, including AIM2, BAK1, CASP3, CHMP4C, CHMP7, GSDMC, GSDME, PYCARD, TNF, and GZMA, are highly expressed in chronic sinusitis.

[0024] like Figure 2 The diagram showing the high expression of pyroptosis-related genes in chronic sinusitis includes... Figure 1 Figure A shows the visualization results of the expression of genes related to pyroptosis. Figure 1 Figure B shows the results of pathway enrichment analysis for genes related to pyroptosis. Figure 1 Figure C shows the results of pathway enrichment analysis for genes related to pyroptosis. Figure 1 Figure D shows the results of pathway enrichment analysis for genes related to pyroptosis. Figure 1 E is a graph showing the results of pathway enrichment analysis of genes related to pyroptosis. Figure 1 F is the graph showing the expression analysis results of pyroptosis-related genes.Figure 1 G is the result chart of pathway enrichment analysis of cell pyroptosis related differential genes, Figure 1 H is the result chart of pathway enrichment analysis of cell pyroptosis related differential genes.

[0025] Example 2 verifies the application of biomarkers in the recurrence of chronic rhinosinusitis

[0026] Based on the cell pyroptosis related genes affecting the recurrence of chronic rhinosinusitis obtained by the above screening, the expression analysis of RCRSwNP group and non-RCRSwNP group found that CASP3 and CHMP7 genes were highly transcribed and expressed in recurrent chronic rhinosinusitis, which were potential recurrence diagnostic markers. The verification process is as follows:

[0027] 1. Sample collection

[0028] (1) Collect the surgically resected nasal mucosa tissue samples of the patients with initial diagnosis of rhinosinusitis, a total of 49 cases.

[0029] (2) Collect the surgically resected nasal mucosa tissue samples of the patients with recurrent rhinosinusitis, a total of 13 cases.

[0030] 2. Detection of pyroptosis related genes

[0031] (1) Transcription of deoxyribonucleic acid (mRNA) extraction

[0032] The tissue mRNA of the nasal mucosa tissue sample is extracted by the traditional method of guanidine isothiocyanate-phenol reagent (trizol) or rapid extraction kit, and its concentration and purity are identified.

[0033] (2) Fluorescent real-time quantitative polymerase chain reaction (qRT-PCR)

[0034] According to the qRT-PCR instructions, the mRNA transcription is carried out, and the transcription expression of pyroptosis related genes CHMP7 and CASP3 is detected. -△△ Ct calculates the relative expression amount of the target gene.

[0035] Select the calculated value of a recurrent patient as a reference, and convert the count values of other patients based on this reference.

[0036] (3) Recurrence risk assessment

[0037] After conversion and analysis, it is found that there is a significant difference in the transcription expression of CASP3 and CHMP7 in the nasal mucosa tissue of the patients with initial diagnosis and recurrence; when the converted value of CASP3 is greater than 0.212 and / or the converted value of CHMP7 is greater than 0.330, it indicates that the patient has a high risk of recurrence after surgery.

[0038] The expression analysis data of each pyroptosis-related gene in the RCRSwNP group and the non-RCRSwNP group are shown in Tables 1-2 below.

[0039] Table 1

[0040]

[0041]

[0042] Table 2

[0043]

[0044]

[0045]

[0046] Specifically, such as Figure 1 The diagram shown illustrates the expression of pyroptosis-related genes in recurrent chronic sinusitis and their role in predicting recurrence. Figure 2 A is a graph showing the expression analysis of AIM2 in the RCRSwNP group and the non-RCRSwNP group. Figure 2 B is a graph showing the expression analysis of BAK1 in the RCRSwNP group and the non-RCRSwNP group. Figure 2 C is the expression analysis diagram of the RCRSwNP group and the non-RCRSwNP group of GZMA. Figure 2 D is the expression analysis diagram of the RCRSwNP group and the non-RCRSwNP group of GSDMC. Figure 2 E is the expression analysis diagram of the GSDME group and the non-RCRSwNP group of GSDMC. Figure 2 F is the expression analysis diagram of the GSDME group and the non-RCRSwNP group in PYCARD. Figure 2 G represents the expression analysis of TNF in the GSDME group and the non-RCRSwNP group. Figure 2 H represents the expression analysis of CHMP4C in the GSDME group and the non-RCRSwNP group. Figure 2 Figure I shows the expression analysis of CASP3 in the GSDME group and the non-RCRSwNP group. Figure 2 J is the expression analysis diagram of the GSDME group and the non-RCRSwNP group of CHMP7. Figure 2 K represents the ROC curve of CASP3. Figure 2 L represents the ROC curve of CHMP7.

[0047] In summary, through bioinformatics analysis and tissue qRT-PCR method verification, it is found that pyroptosis related genes are higher in the expression of the nasal mucosa tissue of the recurrent chronic sinusitis patients than that of the non-recurrent patients; the ROC curve result indicates that the pyroptosis related genes are potential recurrence markers. The present application obtains the nasal mucosa tissue specimen of the patient, and quantitatively detects the pyroptosis related markers by the fluorescence real-time quantitative polymerase chain reaction (qRT-PCR) method, has high specificity and sensitivity in predicting the risk of future recurrence of the chronic sinusitis patient, reminds the patient of the risk of recurrence, causes the patient to pay attention, and makes intervention measures in advance, so as to minimize the risk of disease recurrence as much as possible.

[0048] The above-mentioned embodiments are only preferred embodiments of the present application, and cannot be used to limit the protection scope of the present application, and any non-essential changes and replacements made by those skilled in the art on the basis of the present application shall fall within the protection scope of the present application.

Claims

1. Use of a reagent for detecting the amount of expression of a biomarker, characterized in that, The reagent for detecting the expression amount of the biomarker, the biomarker being CASP3 and / or CHMP7, is applied in the preparation of a kit and / or device for predicting the recurrence of chronic rhinosinusitis with nasal polyps (CRSwNP).

2. Use according to claim 1, characterized in that, The reagent includes a reagent for real-time fluorescent quantitative reverse transcription polymerase chain reaction.

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