CN116741409A - Application of model constructed based on peripheral blood biomarker in predicting upper gastrointestinal tumor chemotherapy efficacy - Google Patents

Abstract

The application provides an application of a model constructed based on peripheral blood biomarkers in predicting upper gastrointestinal tumor chemotherapy efficacy. The peripheral blood biomarker is selected from hemoglobin, eosinophils and CD4 ⁺ T cell, memory CD4 ⁺ T cells, CD8 ⁺ T cells, CD4CD28 ⁺ T cells, CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, CD8CD28 ⁺ One or more of T cells, ELR, leucocytes, platelets, neutrophils and monocytes, wherein the tumor of the upper digestive tract is esophageal cancer, gastric cancer or gastroesophageal junction cancer. The application establishes a prediction model aiming at the upper gastrointestinal tumor chemotherapy curative effect, and the adopted peripheral blood detection index is low in cost, time-saving, convenient for clinical operation and dynamic monitoring, and has important clinical significance and wide applicability.

Description

Model constructed based on peripheral blood biomarkers in predicting chemotherapy for upper gastrointestinal tumors therapeutic applications

This application is Chinese Patent Application No. 202310533252. divisional application.

Technical field

The invention belongs to the field of biotechnology, and particularly relates to the application of a model constructed based on peripheral blood biomarkers in predicting the efficacy of chemotherapy for upper gastrointestinal tumors.

Background technique

Upper gastrointestinal tumors are the most common malignant tumors worldwide, including esophageal cancer, gastric cancer, etc. Surgical resection of early-stage esophageal cancer and gastric cancer may be curative, but only 30-40% of patients have potentially resectable lesions at the time of diagnosis. That is, more than 60% of patients are already in locally advanced stages or have distant metastasis, and the prognosis is poor. For unresectable esophageal cancer and gastric cancer, the current main treatments are chemotherapy, targeted therapy, immune checkpoint inhibitor therapy, etc.; for advanced esophageal cancer and gastric cancer, the efficacy of these treatments is limited. The median survival time of metastatic esophageal cancer and gastric cancer after treatment is only about 10-13 months.

Currently, chemotherapy is still the basic drug for all treatment options for patients with advanced or advanced esophageal cancer and gastric cancer. Currently, the main combination chemotherapy regimen for esophageal cancer is paclitaxel combined with platinum. For gastric cancer, the regimen of oxaliplatin and fluorouracil drugs is more commonly used. How to predict the efficacy of chemotherapy regimens has always been a difficult problem in clinical selection of treatment strategies. Accurately assessing the effectiveness and risk of failure before receiving chemotherapy and identifying the groups who will benefit from chemotherapy are crucial for the treatment of upper gastrointestinal tumors. Currently, there are limited biomarkers for predicting the efficacy of chemotherapy. Some biomarkers for predicting efficacy require detection of cell-free DNA in peripheral blood or invasive tumor biopsy. They also use expensive detection methods, making it difficult to implement dynamic monitoring and have limited clinical application. Therefore, it is of great research significance to select reliable and simple biomarkers or efficacy prediction models.

Contents of the invention

In order to make up for the shortcomings of the existing technology and find reliable and simple biomarkers for predicting upper gastrointestinal tumors (such as esophageal cancer, gastric cancer, gastroesophageal junction cancer) and the efficacy of chemotherapy, the present invention provides a biomarker based on peripheral blood cells and biochemical indicators. The efficacy prediction model jointly established with indicators such as lymphocyte subpopulations and other indicators can provide an economical and time-saving method for predicting the efficacy of chemotherapy for upper gastrointestinal tumors.

A first aspect of the present invention provides the application of a reagent for detecting peripheral blood biomarkers in a sample in preparing a product for predicting and/or evaluating the efficacy of chemotherapy for upper gastrointestinal tumors, and the peripheral blood biomarkers are selected from Hemoglobin, eosinophils, CD4 ⁺ T cells, memory CD4 ⁺ T cells, CD8 ⁺ T cells, CD4CD28 ⁺ T cells, CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, CD8CD28 ⁺ T cells, ELR, white blood cells, platelets, neutrophils One or more types of granulocytes and monocytes.

Further, the upper gastrointestinal tract tumor is esophageal cancer, gastric cancer or gastroesophageal junction cancer.

Preferably, the upper gastrointestinal tract tumor is esophageal squamous cell carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.

Further, the evaluation grade of the chemotherapy efficacy is divided into partial response, disease stabilization and disease progression (refer to Response Evaluation Criteria in Solid Tumors 1.1 (RECISTv1.1)). Of these, partial response and/or stable disease were assessed as achieving disease control.

In one embodiment of the present invention, the chemotherapy regimen for esophageal cancer is a treatment based on paclitaxel combined with platinum drugs.

In one embodiment of the present invention, the chemotherapy regimen for gastric cancer is treatment with oxaliplatin combined with fluorouracil drugs (such as Tegio or capecitabine).

In one embodiment of the invention, hemoglobin, eosinophils, CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, memory CD4 ⁺ T cells, and CD4CD28 ⁺ T cells among the peripheral blood biomarkers can be used as joint indicators. For predicting and/or evaluating the efficacy of chemotherapy for esophageal cancer. Wherein, the higher the content of the peripheral blood biomarker, the easier it is for the therapeutic effect to achieve partial remission and/or disease stabilization (disease control).

In another embodiment of the present invention, eosinophils, CD8 ⁺ T cells, memory CD4 ⁺ T cells, and ELR among the peripheral blood biomarkers can be used as joint indicators to predict and/or evaluate esophageal cancer chemotherapy. Efficacy. Among them, the higher the content of the peripheral blood biomarker, the easier it is for the therapeutic effect to achieve partial remission or even complete remission.

In one embodiment of the present invention, eosinophils, memory CD4 ⁺ T cells, and ELR among the peripheral blood biomarkers can be used as joint indicators to predict and/or evaluate the efficacy of gastric cancer chemotherapy. Wherein, the higher the content of the peripheral blood biomarker, the easier it is for the therapeutic effect to achieve partial remission and/or disease stabilization (disease control).

In another embodiment of the present invention, leukocytes, platelets, neutrophils, monocytes, and CD8CD28 ⁺ T cells among the peripheral blood biomarkers can be used as joint indicators to predict and/or evaluate gastric cancer chemotherapy. Efficacy. Among them, the higher the content of the peripheral blood biomarker, the easier it is for the therapeutic effect to achieve partial remission or even complete remission.

Further, the sample is peripheral blood obtained from a biopsy subject.

Further, the subject is a mammal, especially a human.

Further, the product for predicting and/or evaluating the efficacy of chemotherapy for upper gastrointestinal tumors includes a product for detecting the content of the peripheral blood biomarker.

Further, the product for detecting the content of the peripheral blood biomarker may be a reagent, a kit, a test strip or an instrument platform.

Further, the product for detecting the content of the peripheral blood biomarker can be based on the principles of cell counting technology, for example, resistance method, light scattering method, special cell staining method, microscopic image method, volume conductivity light scattering combined detection method , electrical impedance radio frequency technology combined detection method, light scattering and cell chemistry technology, etc.

A second aspect of the present invention provides a product for predicting and/or evaluating the efficacy of chemotherapy for upper gastrointestinal tumors. The product for predicting and/or evaluating the efficacy of chemotherapy for upper gastrointestinal tumors includes detecting peripheral blood as described in the first aspect. Biomarker content of the product.

Further, the upper gastrointestinal tract tumor is esophageal cancer, gastric cancer or gastroesophageal junction cancer.

Preferably, the upper gastrointestinal tract tumor is esophageal squamous cell carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.

Further, the evaluation grade of the chemotherapy efficacy is divided into partial response, disease stability and disease progression. Of these, partial response and/or stable disease were assessed as achieving disease control levels.

Further, the product for detecting the content of the peripheral blood biomarker may be a reagent, a kit, a test strip or an instrument platform.

Further, the product for detecting the content of the peripheral blood biomarker can be based on the principles of cell counting technology, for example, resistance method, light scattering method, special cell staining method, microscopic image method, volume conductivity light scattering combined detection method , electrical impedance radio frequency technology combined detection method, light scattering and cell chemistry technology, etc.

A third aspect of the present invention provides a prediction model for the efficacy of chemotherapy for upper gastrointestinal tumors. The prediction model uses the peripheral blood biomarkers described in the first aspect as variables.

Further, the upper gastrointestinal tract tumor is esophageal cancer, gastric cancer or gastroesophageal junction cancer.

Preferably, the upper gastrointestinal tract tumor is esophageal squamous cell carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.

Further, the evaluation grade of the chemotherapy efficacy is divided into partial response, disease stability and disease progression. Of these, partial response and/or stable disease were assessed as achieving disease control.

In one embodiment of the invention, the variables of the prediction model for the efficacy of chemotherapy for esophageal cancer are a combination of hemoglobin, eosinophils, CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, memory CD4 ⁺ T cells and CD4CD28 ⁺ T cells .

In another embodiment of the present invention, the variables of the prediction model for the efficacy of chemotherapy for esophageal cancer are a combination of eosinophils, CD8 ⁺ T cells, memory CD4 ⁺ T cells and ELR.

In one embodiment of the present invention, the variables of the prediction model for gastric cancer chemotherapy efficacy are a combination of eosinophils, memory CD4 ⁺ T cells and ELR.

In another embodiment of the present invention, the variables of the prediction model for the efficacy of chemotherapy for gastric cancer are a combination of white blood cells, platelets, neutrophils, monocytes and CD8CD28 ⁺ T cells.

A fourth aspect of the present invention provides a method for predicting and/or evaluating the efficacy of chemotherapy for upper gastrointestinal tumors, which includes the step of detecting a reagent for the peripheral blood biomarker described in the first aspect.

Further, the upper gastrointestinal tract tumor is esophageal cancer, gastric cancer or gastroesophageal junction cancer.

Preferably, the upper gastrointestinal tract tumor is esophageal squamous cell carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.

Further, the evaluation grade of the chemotherapy efficacy is divided into partial response, disease stability and disease progression. Of these, partial response and/or stable disease were assessed as achieving disease control levels.

Further, the method includes the following steps:

(1) Obtain subject samples;

(2) Detect the content of the peripheral blood biomarkers in the subject’s sample;

(3) Correlate the measured content of the peripheral blood biomarker with the chemotherapy efficacy of the subject's upper gastrointestinal tumors (such as esophageal cancer, gastric cancer, gastroesophageal junction adenocarcinoma).

Further, the sample is peripheral blood obtained from a biopsy subject.

Further, the subject is a mammal, especially a human.

Further, the standard for predicting and/or evaluating the efficacy of chemotherapy for upper gastrointestinal tumors is: detecting the content of the peripheral blood biomarker, and comparing the subject's peripheral blood biomarker before chemotherapy with the reference The higher the content, the easier it is for the chemotherapy effect of upper gastrointestinal tumors to reach the level of disease control (such as partial remission and/or disease stabilization), and vice versa.

Further, the reference is the content value corresponding to the maximum Youden index of the peripheral blood biomarker in the subject before chemotherapy.

In one embodiment of the invention, the higher the contents of hemoglobin, eosinophils, CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, memory CD4 ⁺ T cells, and CD4CD28 ⁺ T cells, the better the chemotherapy efficacy of esophageal cancer. Partial remission and/or disease stabilization (disease control) can be easily achieved.

In another embodiment of the present invention, the higher the contents of eosinophils, CD8 ⁺ T cells, memory CD4 ⁺ T cells, and ELR, the easier it is to achieve partial remission of esophageal cancer through chemotherapy.

In one embodiment of the present invention, the higher the content of eosinophils, memory CD4 ⁺ T cells, and ELR, the easier it is for the chemotherapy efficacy of gastric cancer to achieve partial remission and/or disease stabilization (disease control).

In another embodiment of the present invention, the higher the content of leukocytes, platelets, neutrophils, monocytes, and CD8CD28 ⁺ T cells, the easier it is for the chemotherapeutic effect of gastric cancer to achieve partial remission.

The present invention provides a peripheral blood biomarker for predicting the efficacy of chemotherapy for upper gastrointestinal tumors (such as esophageal cancer, gastric cancer, and gastroesophageal junction cancer). It can be used for auxiliary diagnosis in predicting the risk of chemotherapy for upper gastrointestinal tumors, and has good detection specificity. ,high sensitivity. The peripheral blood index detection used in the present invention is all conventional kits, which are low-cost, time-saving, convenient for clinical operation and dynamic monitoring, and have important clinical significance and wide applicability. The peripheral blood joint indicators of the present invention respectively reflect that some inflammatory factors, peripheral blood cells and lymphocyte subpopulations are closely related to the development of tumors, and interact with the immune microenvironment of the body, and are expected to be used for monitoring the course of upper gastrointestinal tumors, Evaluation indicators for prediction and prognosis.

Description of drawings

Figure 1 shows the ROC curve of combined indicator model A to predict the efficacy of chemotherapy for esophageal cancer to achieve disease control.

Figure 2 shows the ROC curve of combined indicator model B to predict the efficacy of chemotherapy for esophageal cancer to achieve partial remission of the disease.

Figure 3 shows the ROC curve of combined indicator model C to predict the efficacy of chemotherapy for gastric cancer to achieve disease control.

Figure 4 shows the ROC curve of the combined indicator model D to predict the efficacy of gastric cancer chemotherapy to achieve partial remission of the disease.

Detailed ways

Unless otherwise defined, all scientific and technical terms used in the present invention have the same meaning as commonly understood by a person skilled in the art to which this invention relates.

The term "Youden index" is a method of evaluating the authenticity of a screening test and represents the overall ability of the screening method to detect true patients and non-patients. The Youden index is the sum of sensitivity and specificity minus 1. The value corresponding to the cutoff point at the maximum Youden index is used as the CUT OFF value to ensure the optimal accuracy of the kit.

The embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. If the specific conditions are not specified in the examples, the conditions should be carried out according to the conventional conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional products that can be purchased commercially.

Example 1

1.1 Research objects

A total of 92 patients with upper gastrointestinal tumors (38 cases of esophageal cancer and 54 cases of gastric cancer) were prospectively enrolled in this study.

Inclusion criteria: (1) Aged over 18 years old; (2) Esophageal squamous cell carcinoma, gastric or gastroesophageal junction adenocarcinoma confirmed by histopathology or cytology; (3) Locally advanced or stage IV at the time of enrollment (AJCC 7.0); (4) Have not received surgery or other anti-tumor treatment within 6 months before enrollment; (5) Organ function is normal; (6) Meet the requirements for chemical drug treatment; (7) Voluntarily participate in this study.

1.2 Research methods

1.2.1 Treatment plan and efficacy evaluation

Clinical researchers choose standard chemotherapy regimens: esophageal squamous cell carcinoma is mainly treated with paclitaxel combined with platinum; for gastric and gastroesophageal junction adenocarcinoma, oxaliplatin combined with fluorouracil-based drug regimen (tegio or capecitabine) is mainly used.

Efficacy evaluation: Refer to Response Evaluation Criteria in Solid Tumors 1.1 (RECIST v1.1), and conduct evaluation of efficacy every 6 weeks; efficacy evaluation grades are divided into: partial response (PR), stable disease (SD), disease Progression (PD); both partial response and stable disease were assessed as achieving disease control.

1.2.2 Test items

Peripheral blood examination was performed within 1 week before the patient's anti-tumor treatment (baseline): complete blood cell analysis, lymphocyte subsets, erythrocyte sedimentation rate (ESR), hypersensitive C-reactive protein (hsCRP) ), albumin ALB/prealbumin preALB, lymphocyte subsets and other testing items. The combined marker levels (MLR, NLR, ELR, BLR, PLR, CAR, COP, CLR, CBR) were calculated separately for each patient.

MLR = number of monocytes/number of lymphocytes; NLR = number of neutrophils/number of lymphocytes; ELR = number of eosinophils/number of lymphocytes; BLR = number of basophils/number of lymphocytes; PLR = number of platelets/ Number of lymphocytes; CAR=CRP/ALB; COP=CRP/preALB; CLR=CRP/lymphocytes; CBR=CRP/BMI.

1.3 Statistical analysis

SPSS26.0 was used for statistical analysis, T test and ANOVA were used to compare data differences between groups, and Mann-Whitney and Kruskal-Wallis tests were used for non-parametric tests. Kaplan-Meier survival analysis was used to compare differences in survival, and ROC analysis was used to determine the effectiveness of marker differences as diagnostic criteria. Setting two-way P<0.05 indicates that the difference is statistically significant. GraphPad Prism 8.0 software (San Diego, USA) was used for statistical analysis and graphics, and Mann-Whitney and Kruskal-Wallis rank sum tests were used to compare differences in clinical data.

1.4 Clinical characteristics of enrolled patients

Table 1 Clinical characteristics of patients with esophageal cancer and gastric cancer

Table 2 Circulation parameters of patients with esophageal cancer and gastric cancer

1.5 Single indicator screening

Table 3 Esophageal cancer indicator screening results

Table 4 Gastric cancer indicator screening results

As shown in Table 3, the single-index screening for early stage esophageal cancer chemotherapy to achieve disease control found that hemoglobin (Hb), eosinophils (EOS), CD4CD28 ⁺ T/CD4 ⁺ T cells, memory CD4 ⁺ T, and CD4CD28 ⁺ T cells had Significant difference; single-index screening of partial response achieved by early-stage esophageal cancer chemotherapy found that there were significant differences in eosinophil (EOS) levels, CD8 ⁺ T cell levels, memory CD4 ⁺ T, and ELR (P < 0.05).

As shown in Table 4, the single-index screening for early-stage gastric cancer chemotherapy to achieve disease control found that eosinophils (EOS), memory CD4 ⁺ T, and ELR were significantly different; the single-index screening for early-stage gastric cancer to achieve partial remission through chemotherapy found that white blood cells ( There were significant differences in WBC) levels, platelet (PLT) levels, neutrophils (NEUT), monocytes (Mono), and CD8CD28 ⁺ T cells (P<0.05).

Example 2 Screening peripheral blood markers related to the efficacy of chemotherapy for esophageal cancer and building a prediction model

Through early single-index screening, it was found that the higher the hemoglobin (Hb), eosinophils (EOS), CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, memory CD4 ⁺ T, and CD4CD28 ⁺ T cell count, the easier it is for esophageal cancer chemotherapy to reach the disease Control (partial remission + stable disease), the less likely it is to progress.

The above single indicators with predictive value for therapeutic efficacy were combined to construct a therapeutic efficacy prediction model, and their predictive effects were evaluated through ROC diagnostic analysis. The results showed that their area under the receiver operating characteristic curve (AUROC) was between 0.4210 and 0.7920.

The area under the AUC curve of joint index model A to predict the efficacy of chemotherapy for esophageal cancer to achieve disease control (partial response PR + stable disease SD) is 79.2% (Figure 1), the sensitivity is 82.6%, the specificity is 80%, p=0.012 (95CI 0.609 -0.975), which has better prediction performance than a single indicator.

It was also found that the higher the eosinophil (EOS) level, CD8 ⁺ T cell level, memory CD4 ⁺ T count, and ELR, the easier it is to achieve partial remission (PR). Single indicators with predictive value for therapeutic effect (PR) were combined to construct a prediction model for therapeutic effect, and their predictive effects were evaluated through ROC diagnostic analysis. The results showed that their area under the receiver operating characteristic curve (AUROC) ranged from 0.701 to 0.851. between.

The area under the AUC curve of the combined index model B to predict the efficacy of chemotherapy for esophageal cancer to reach partial response (PR) was 85.1% (Figure 2), the sensitivity was 77.8%, the specificity was 89.7%, p=0.002 (95CI 0.691-1.000), It has better prediction performance than a single indicator.

Example 3 Screening peripheral blood markers related to the efficacy of chemotherapy for gastric cancer and building a prediction model

Through early single indicator screening, it was found that the higher the eosinophils (EOS), memory CD4 ⁺ T, and ELR, the easier it is for gastric cancer chemotherapy to achieve disease control (partial response + disease stability), and the less likely it is to progress.

The above single indicators with predictive value for therapeutic efficacy were combined to construct a therapeutic efficacy prediction model, and their predictive effects were evaluated through ROC diagnostic analysis. The results showed that their area under the receiver operating characteristic curve (AUROC) ranged from 0.722 to 0.936.

The area under the AUC curve of the combined index model C to predict the efficacy of gastric cancer chemotherapy to achieve disease control (partial response PR + stable disease SD) is 93.6% (Figure 3), the sensitivity is 84.1%, the specificity is 90%, p<0.001 (95CI 0.861- 1.000), which has better prediction performance than a single indicator.

The higher the white blood cell (WBC) level, platelet (PLT) level, neutrophil (NEUT) count, monocyte (Mono), and CD8CD28 ⁺ T cells, the easier it is to achieve partial remission (PR).

Single indicators with predictive value for efficacy (PR) were combined to construct a efficacy prediction model, and their predictive effects were evaluated through ROC diagnostic analysis. The results showed that the AUROC ranged from 0.671 to 0.807.

The area under the AUC curve of the combined index model D to predict the efficacy of gastric cancer chemotherapy to achieve partial response (PR) was 80.7% (Figure 4), the sensitivity was 88.9%, the specificity was 66.7%, p<0.001 (95CI 0.692-0.922), compared with Single indicator prediction performance is better.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention, but not to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: It is still possible to modify the technical solutions recorded in the foregoing embodiments, or to equivalently replace some or all of the technical features; and these modifications or substitutions do not deviate from the essence of the corresponding technical solutions from the technical solutions of the embodiments of the present invention. range.

Claims (4)

1. The application of a reagent for detecting peripheral blood biomarkers in a sample in the preparation of a product for predicting and/or evaluating the chemotherapeutic effect of upper gastrointestinal tumor is characterized in that the peripheral blood biomarkers are hemoglobin, eosinophils and CD4CD28 ⁺ T/CD4 ⁺ T cell ratio, memory CD4 ⁺ T cells and CD4CD28 ⁺ T cells, the upper gastrointestinal tumor is esophageal cancer;

or, the peripheral blood biomarker is eosinophil and memory CD4 ⁺ T cells and ELR, the upper gastrointestinal tumor being gastric cancer;

or, the peripheral blood biomarker is leukocyte, platelet, neutrophil, monocyte and CD8CD28 ⁺ T cells, the upper gastrointestinal tumor is gastric cancer.

2. The use of claim 1, wherein the sample is peripheral blood obtained from a biopsy subject.

3. The use according to claim 1, wherein said product for predicting and/or assessing the efficacy of chemotherapy of tumors of the upper digestive tract comprises a product for detecting the content of said peripheral blood biomarkers.

4. The use according to claim 3, wherein the product for detecting the content of the peripheral blood biomarker is a reagent, a kit, a test paper or an instrument platform.