CN116246795B - Application of reagent for detecting peripheral blood biomarkers in sample in preparation of product for predicting curative effect of esophageal cancer chemotherapy - Google Patents
Application of reagent for detecting peripheral blood biomarkers in sample in preparation of product for predicting curative effect of esophageal cancer chemotherapy Download PDFInfo
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Abstract
The invention 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 invention 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
Technical Field
The invention belongs to the technical field of biology, and particularly relates to application of a model constructed based on peripheral blood biomarkers in predicting upper gastrointestinal tumor chemotherapy efficacy.
Background
Upper gastrointestinal tumors are the most common malignant tumors worldwide, including esophageal cancer, gastric cancer, and the like. Early esophageal cancer and gastric cancer can be cured by surgical excision, but only 30-40% of patients are potential resectable lesions when in a doctor, namely more than 60% of patients are in local advanced stage or have distant metastasis, and the prognosis is poor. For unresectable esophageal cancer and gastric cancer, the main treatment methods at present are chemotherapy, targeted therapy, immune checkpoint inhibitor therapy and the like; for advanced esophageal cancer and gastric cancer, the curative effect of the treatment means is limited. The median survival time of metastatic esophageal cancer and gastric cancer after treatment is only about 10-13 months.
At present, for patients with advanced or advanced esophageal cancer, gastric cancer, chemotherapy remains the basis for all treatment regimens. The main combined chemotherapy scheme at present is paclitaxel combined platinum scheme aiming at esophageal cancer, and oxaliplatin and fluorouracil medicament scheme aiming at gastric cancer are more selected. How to predict the efficacy of a chemotherapy regimen is always a difficult problem in selecting a treatment strategy clinically, and accurately evaluates the effectiveness of the treatment and the risk of failure before receiving the chemotherapy, and makes clear that the population benefited from the chemotherapy is critical for the treatment of upper gastrointestinal tumors. The existing biomarkers for predicting the curative effect of chemotherapy are limited, partial curative effect prediction markers need to be detected by peripheral blood free DNA or invasive tumor biopsy, and dynamic monitoring is difficult to implement by adopting an expensive detection means, so that the clinical application is limited. Therefore, the selection of reliable and simple biomarkers or efficacy prediction models has important research significance.
Disclosure of Invention
In order to make up for the deficiency of the prior art, find reliable, simple and convenient and predict upper digestive tract tumor (such as esophagus cancer, stomach esophagus juncture cancer) and relevant biomarker of chemotherapy curative effect, the invention provides the curative effect prediction model based on index combination establishment such as peripheral blood cell, biochemical index and lymphocyte subgroup, etc., can be well aimed at upper digestive tract tumor curative effect, provide an economic, time-saving method to predict.
In a first aspect, the invention provides the use of a reagent for detecting a peripheral blood biomarker selected from the group consisting of hemoglobin, eosinophils, and CD4 in a sample in the manufacture of a product for predicting and/or assessing the efficacy of chemotherapy in tumors of the upper digestive tract + T cell, 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, monocytes or combinations thereof.
Further, the tumor of the upper digestive tract is esophageal cancer, gastric cancer or gastroesophageal junction cancer.
Preferably, the tumor of the upper digestive tract is esophageal squamous carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.
Further, the evaluation grade of the chemotherapeutic effect is classified into partial remission, disease stabilization and disease progression (refer to solid tumor effect evaluation criteria 1.1 (Response Evaluation Criteria in Solid Tumors 1.1, recist v 1.1)). Wherein partial remission and/or disease stability is assessed as achieving disease control.
In one embodiment of the invention, the chemotherapeutic regimen for esophageal cancer is a paclitaxel-combined platinum-based therapy.
In one embodiment of the invention, the chemotherapeutic regimen for gastric cancer is treatment with oxaliplatin in combination with fluorouracil (e.g., tegafur or capecitabine).
In one embodiment of the invention, the peripheral blood biomarker comprises hemoglobin, eosinophils, and CD4CD28 + T/CD4 + T cell ratio, memory CD4 + T cells, CD4CD28 + T cells can be used as a combined indicator to predict and/or assess the efficacy of chemotherapy for esophageal cancer. Wherein, the higher the content of the peripheral blood biomarker, the more easily the curative effect can achieve partial remission and/or disease stabilization (disease control).
In another embodiment of the present invention, the eosinophils, CD8, in the peripheral blood biomarker + T cell, memory CD4 + T cells and ELR can be used as combined indexes for predicting and/or evaluating the curative effect of esophagus cancer chemotherapy. Wherein, the higher the content of the peripheral blood biomarker, the more easily the curative effect is partially or even completely relieved.
In one embodiment of the invention, eosinophils, memory CD4, in the peripheral blood biomarker + T cells and ELR can be used as combined indexes for predicting and/or evaluating the curative effect of gastric cancer chemotherapy. Wherein, the higher the content of the peripheral blood biomarker, the more easily the curative effect can achieve partial remission and/or disease stabilization (disease control).
In another embodiment of the present invention, the peripheral blood biomarker is selected from the group consisting of white blood cells, platelets, neutrophils, monocytes, and CD8CD28 + T cells can be used as conjunctive fingersThe marker is used for predicting and/or evaluating the curative effect of gastric cancer chemotherapy. Wherein, the higher the content of the peripheral blood biomarker, the more easily the curative effect is partially or even completely relieved.
Further, the sample is peripheral blood obtained from a biopsy subject.
Further, the subject is a mammal, particularly a human.
Further, the product for predicting and/or evaluating the efficacy of chemotherapy on upper gastrointestinal tumors comprises a product for detecting the content of the peripheral blood biomarker.
Further, the product for detecting the content of the peripheral blood biomarker can be a reagent, a kit, test paper or an instrument platform.
Further, the product for detecting the content of the peripheral blood biomarker can be based on the principle of cell counting technology, such as a resistance method, a light scattering method, a special cell staining method, a microscopic image method, a volume conductance light scattering combined detection method, an electrical impedance radio frequency technology combined detection method, a light scattering and cytochemistry technology and the like.
In a second aspect of the invention, there is provided a product for predicting and/or assessing the efficacy of chemotherapy of a tumor in the upper digestive tract, said product comprising a product for detecting the level of said peripheral blood biomarker of the first aspect.
Further, the tumor of the upper digestive tract is esophageal cancer, gastric cancer or gastroesophageal junction cancer.
Preferably, the tumor of the upper digestive tract is esophageal squamous carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.
Further, the assessment of the efficacy of chemotherapy is classified as partial remission, disease stabilization and disease progression. Wherein partial remission and/or disease stability is assessed as reaching a level of disease control.
Further, the product for detecting the content of the peripheral blood biomarker can be a reagent, a kit, test paper or an instrument platform.
Further, the product for detecting the content of the peripheral blood biomarker can be based on the principle of cell counting technology, such as a resistance method, a light scattering method, a special cell staining method, a microscopic image method, a volume conductance light scattering combined detection method, an electrical impedance radio frequency technology combined detection method, a light scattering and cytochemistry technology and the like.
In a third aspect of the present invention, there is provided a predictive model of the efficacy of chemotherapy for tumors of the upper digestive tract, said predictive model having as variables said peripheral blood biomarkers of the first aspect.
Further, the tumor of the upper digestive tract is esophageal cancer, gastric cancer or gastroesophageal junction cancer.
Preferably, the tumor of the upper digestive tract is esophageal squamous carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.
Further, the assessment of the efficacy of chemotherapy is classified as partial remission, disease stabilization and disease progression. Wherein partial remission and/or disease stability is assessed as achieving disease control.
In one embodiment of the invention, the variables of the predictive model of the efficacy of chemotherapy for esophageal cancer are hemoglobin, eosinophils, CD4CD28 + T/CD4 + T cell ratio, memory CD4 + T cells and CD4CD28 + T cell combinations.
In another embodiment of the present invention, the variables of the predictive model of the efficacy of chemotherapy for esophageal cancer are eosinophils, CD8 + T cell, memory CD4 + T cell and ELR combinations.
In one embodiment of the invention, the variables of the predictive model of efficacy of gastric cancer chemotherapy are eosinophils, memory CD4 + T cell and ELR combinations.
In another embodiment of the present invention, the variables of the predictive model of efficacy of gastric cancer chemotherapy are white blood cells, platelets, neutrophils, monocytes and CD8CD28 + T cell combinations.
In a fourth aspect of the invention, there is provided a method of predicting and/or assessing the efficacy of chemotherapy for tumors of the upper digestive tract comprising the step of detecting an agent of the peripheral blood biomarker of the first aspect.
Further, the tumor of the upper digestive tract is esophageal cancer, gastric cancer or gastroesophageal junction cancer.
Preferably, the tumor of the upper digestive tract is esophageal squamous carcinoma, gastric adenocarcinoma or gastroesophageal junction adenocarcinoma.
Further, the assessment of the efficacy of chemotherapy is classified as partial remission, disease stabilization and disease progression. Wherein partial remission and/or disease stability is assessed as reaching a level of disease control.
Further, the method comprises the following steps:
(1) Obtaining a subject sample;
(2) Detecting the content of the peripheral blood biomarker in a sample of the subject;
(3) And correlating the measured content of the peripheral blood biomarker with the chemotherapeutic effect of the tumor of the upper digestive tract (such as esophageal cancer, gastric cancer and adenocarcinoma at the boundary of the stomach and the esophagus) of the subject.
Further, the sample is peripheral blood obtained from a biopsy subject.
Further, the subject is a mammal, particularly a human.
Further, the criteria for predicting and/or assessing the efficacy of chemotherapy for upper gastrointestinal tumors are: the higher the level of the peripheral blood biomarker prior to chemotherapy in the subject as compared to the reference, is an indication that the higher the level of disease control (e.g., partial remission and/or disease stabilization) is for the upper gastrointestinal tumor chemotherapy efficacy, and vice versa.
Further, the reference is a content value corresponding to a maximum approximate dengue index of the peripheral blood biomarker prior to chemotherapy of the subject.
In one embodiment of the invention, the haemoglobin, eosinophil, CD4CD28 + T/CD4 + T cell ratio, memory CD4 + T cells, CD4CD28 + The higher the content of T cells, the more easily the chemotherapy effect of esophageal cancer is achievedPartial remission and/or disease stabilization (disease control).
In another embodiment of the present invention, the eosinophil, CD8 + T cell, memory CD4 + The higher the content of T cells and ELR, the more easily the chemotherapy effect of the esophageal cancer is partially relieved.
In one embodiment of the invention, the eosinophil, memory CD4 + The higher the content of T cells and ELR, the more easily the chemotherapy effect of gastric cancer can achieve partial remission and/or disease stabilization (disease control).
In another embodiment of the present invention, the said leukocytes, platelets, neutrophils, monocytes, CD8CD28 + The higher the content of T cells, the more easily the chemotherapy effect of gastric cancer is partially relieved.
The peripheral blood biomarker for predicting the chemotherapy efficacy of the upper gastrointestinal tumor (such as esophageal cancer, gastric cancer and gastroesophageal junction cancer) can be used for the auxiliary diagnosis of predicting the chemotherapy risk of the upper gastrointestinal tumor, and has the advantages of good detection specificity and high sensitivity. The peripheral blood index detection adopted by the invention is a conventional kit, has low cost and time saving, is convenient for clinical operation and dynamic monitoring, and has important clinical significance and wide applicability. The peripheral blood combined index of the invention reflects that part of inflammatory factors, peripheral blood cells and lymphocyte sub-populations are closely related to the development of tumors, and have interaction with the immune microenvironment of the organism, thus being hopeful to be used as an evaluation index for monitoring, predicting and prognosis judging the tumor disease course of the upper digestive tract.
Drawings
Fig. 1 shows ROC curves for predicting the curative effect of esophageal cancer chemotherapy to achieve disease control by the combined index model a.
Fig. 2 shows ROC curves for predicting the efficacy of esophageal cancer chemotherapy to achieve partial remission of the disease in combination with index model B.
Fig. 3 shows ROC curves for predicting the efficacy of gastric cancer chemotherapy to achieve disease control in combination with index model C.
Fig. 4 shows ROC curves for predicting the efficacy of gastric cancer chemotherapy to achieve partial remission of the disease in combination with index model D.
Detailed Description
Unless defined otherwise, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention relates.
The term "about log index" is a method of evaluating the authenticity of a screening test and means the total ability of the screening method to find true patients and non-patients. The about log index is the sum of sensitivity and specificity minus 1. The corresponding value of the CUT-OFF point corresponding to the maximum approximate dengue index is used as the CUT OFF value, so that the optimal accuracy of the kit can be ensured.
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1
1.1 study object
92 patients with upper gastrointestinal tumor (38 cases of esophageal cancer and 54 cases of gastric cancer) were prospective into the group of the study.
Group entry criteria: (1) older than 18 years; (2) Determining diagnosis of esophageal squamous carcinoma, stomach or gastroesophageal junction adenocarcinoma by histopathology or cytology; (3) The group entering time stage is local late stage or IV stage (AJCC 7.0); (4) No surgery or other anti-tumor treatment was received within 6 months prior to group entry; (5) normal organ function; (6) meets the requirements of chemical drug treatment; (7) voluntary participation in the study.
1.2 research methods
1.2.1 treatment regimens and efficacy assessment
Clinical researchers selected standard chemotherapy regimens: esophageal squamous carcinoma is mainly formed by paclitaxel and platinum; the stomach and gastroesophageal junction adenocarcinoma is mainly selected from oxaliplatin and fluorouracil drug scheme (tegafur or capecitabine).
Efficacy evaluation: efficacy assessment is performed every 6 weeks with reference to efficacy assessment criteria 1.1 for solid tumors (Response Evaluation Criteria in Solid Tumors.1, recist v 1.1); the efficacy evaluation grade fraction is: partial Remission (PR), disease Stabilization (SD), disease Progression (PD); both partial remission and disease stability were assessed as achieving disease control.
1.2.2 detection items
Peripheral blood studies were left 1 week (baseline) prior to anti-tumor treatment of the patient: whole blood cell analysis, lymphocyte subpopulation, erythrocyte sedimentation rate (erythrocyte sedimentation rate, ESR), hypersensitive C-reactive protein (hsCRP), albumin ALB/prealbumin, lymphocyte subpopulation, and the like. The combined marker levels were calculated for each patient separately (MLR, NLR, ELR, BLR, PLR, CAR, COP, CLR, CBR).
MLR = number of monocytes/number of lymphocytes; NLR = neutrophil count/lymphocyte count; ELR = eosinophil number/lymphocyte number; BLR = basophil number/lymphocyte number; PLR = platelet number/lymphocyte number; car=crp/ALB; COP = CRP/preALB; CLR = CRP/lymphocyte; CBR = CRP/BMI.
1.3 statistical analysis
Statistical analysis was performed using SPSS 26.0, data differences between groups were compared using T-test, ANOVA analysis of variance, and non-parametric tests were performed using Mann-Whitney and Kruskal-Wallis tests. Comparing the survival differences using Kaplan-Meier survival analysis, and judging the effectiveness of the marker differences as diagnostic criteria using ROC analysis. The difference was statistically significant by setting bi-directional P < 0.05. Statistical analysis and mapping was performed using GraphPad Prism 8.0 software (San Diego, USA) and clinical data differences were compared using Mann-Whitney and Kruskal-Wallis rank sum test.
1.4 clinical characterization of patients in group
TABLE 1 clinical characteristics of patients with esophageal cancer and gastric cancer
TABLE 2 circulation parameters for patients with esophageal cancer and gastric cancer
1.5 Single index screening
TABLE 3 screening results for esophageal cancer index
TABLE 4 gastric cancer index screening results
As shown in Table 3, single index screening for disease control achieved by pre-esophageal cancer chemotherapy revealed hemoglobin (Hb), eosinophils (EOS), CD4CD28 + T/CD4 + T cell, memory CD4 + T、CD4CD28 + T cells have significant differences; single index screening for partial remission of early stage esophageal cancer chemotherapy found Eosinophil (EOS) levels, CD8 + T cell level, memory CD4 + T, ELR has significant differences (P < 0.05).
As shown in Table 4, single index screening for disease control achieved by pre-gastric cancer chemotherapy revealed Eosinophils (EOS), memory CD4 + T, ELR has significant differences; single index screening to achieve partial remission of pre-gastric cancer chemotherapy found that leukocyte (WBC) levels, platelet (PLT) levels, neutrophils (NEUT), monocytes (Mono), CD8CD28 + T cells have significant differences (P < 0.05).
Example 2 screening of peripheral blood markers related to efficacy of chemotherapy for esophageal cancer, construction of predictive model
By early single index screening, hemoglobin (Hb), eosinophils (EOS), CD4CD28 were found + T/CD4 + T cell ratio, memory CD4 + T、CD4CD28 + The higher the T cell count, the easier the esophageal cancer chemotherapy achieves disease control (partial remission+disease stabilization), the moreIs not easy to progress.
The single indexes with efficacy prediction values are combined to construct an efficacy prediction model, and the prediction effects of the single indexes are evaluated through ROC diagnosis analysis, so that the Area (AUROC) of the single indexes under the working characteristic curve of a receiver is 0.4210-0.7920.
The combined index model A predicts that the curative effect of esophageal cancer chemotherapy reaches 79.2% of the area under an AUC curve of disease control (partial remission PR+stable SD) (figure 1), the sensitivity is 82.6%, the specificity is 80%, and the p=0.012 (95 CI 0.609-0.975), so that the prediction efficiency is better than that of single index.
At the same time, eosinophil (EOS) level, CD8 + T cell level, memory CD4 + The higher the T count, the easier it is to achieve Partial Remission (PR). The single indexes with efficacy (PR) predictive value are combined to construct an efficacy prediction model, and the prediction effect of the efficacy prediction model is evaluated through ROC diagnosis analysis, so that the result shows that the Area (AUROC) of the efficacy prediction model under the working characteristic curve of the receiver is between 0.701 and 0.851.
The combined index model B predicts that the curative effect of esophageal cancer chemotherapy reaches 85.1% of the area under an AUC curve of partial remission of disease (PR) (figure 2), the sensitivity is 77.8%, the specificity is 89.7%, and the p=0.002 (95 CI 0.691-1.000), and the predicted efficacy is better than that of single index.
Example 3 screening of peripheral blood markers related to efficacy of gastric cancer chemotherapy and construction of predictive model
Eosinophils (EOS), memory CD4 were found by pre-single index screening + The higher T, ELR, the easier the gastric cancer chemotherapy achieves disease control (partial remission+stable disease), the less likely it is to progress.
The single indexes with efficacy prediction values are combined to construct an efficacy prediction model, and the prediction effects of the single indexes are evaluated through ROC diagnosis analysis, so that the Area (AUROC) of the single indexes under the working characteristic curve of a receiver is between 0.722 and 0.936.
The combined index model C predicts that the curative effect of gastric cancer chemotherapy reaches 93.6% of the area under an AUC curve of disease control (partial remission of PR+ disease stable SD) (figure 3), the sensitivity is 84.1%, the specificity is 90%, and the p is less than 0.001 (95 CI 0.861-1.000), so that the predicted efficacy is better than that of single index.
White Blood Cell (WBC) level, platelet (PLT) level, neutrophil count (NEUT), monocyte (Mono), CD8CD28 + The higher the T cells, the easier it is to achieve Partial Remission (PR).
Single indexes with efficacy (PR) predictive value are combined to construct an efficacy prediction model, and the prediction effect of the efficacy prediction model is evaluated through ROC diagnosis analysis, so that AUROC is between 0.671 and 0.807.
The combined index model D predicts that the curative effect of gastric cancer chemotherapy reaches 80.7% of the area under an AUC curve of partial remission of disease (PR) (figure 4), the sensitivity is 88.9%, the specificity is 66.7%, and the p <0.001 (95 CI 0.692-0.922) has better prediction efficiency than the single index.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
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 esophageal cancer is characterized in that the peripheral blood biomarkers are eosinophils and CD8 + T cell, memory CD4 + T cells and ELR.
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 esophageal cancer comprises a product for detecting the level of said peripheral blood biomarker.
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.
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