Prediction model construction and clinical application of biomarker based on breast cancer adriamycin chemotherapy induced cardiotoxicity

Abstract

The invention belongs to the technical field of medical information, and particularly relates to prediction model construction and clinical application of a biomarker based on breast cancer adriamycin chemotherapy induced cardiotoxicity. The invention determines that the FOXD3 gene and / or the expression product thereof, the GBX2 gene and / or the expression product thereof, the SOX3 gene and / or the expression product thereof, or the HIST1H4 gene and / or the expression product thereof are obviously related to the doxorubicin-induced cardiotoxicity. Detection of an FOXD3 gene and / or an expression product thereof, a GBX2 gene and / or an expression product thereof, an SOX3 gene and / or an expression product thereof, or an HIST1H4 gene and / or an expression product thereof in serum is combined with whole blood SIRI index detection to construct a column diagram prediction model, so that the occurrence of doxorubicin-induced cardiotoxicity can be intuitively and accurately predicted; conditions are provided for early heart function correction and permanent heart damage avoiding of breast cancer patients who use adriamycin for chemotherapy.

Description

Technical Field

[0001] The present invention relates to the field of medical information technology, and in particular to the construction and clinical application of a prediction model based on biomarkers of doxorubicin chemotherapy-induced cardiotoxicity in breast cancer. Background Art

[0002] Adriamycin (ADM) is an anthracycline antibiotic with a broad anti-tumor spectrum and strong effects. It is one of the most commonly used anti-tumor drugs. Adriamycin is also a common anti-tumor drug for breast cancer patients, and its efficacy has been confirmed by a large number of clinical studies. Studies have shown that adriamycin can cause cardiotoxicity, and the characteristics of cardiotoxicity are cumulative dose-dependent. Evaluating the patient's cardiac function during early chemotherapy or correcting cardiac function early can avoid permanent heart damage.

[0003] FOXD3 is a transcription factor belonging to the FH superfamily. It was first discovered in embryonic stem cells and is essential for neural crest development and stem cell biological processes. It can maintain the multipotent properties of inner cell mass, neural crest and trophoblast progenitor cells. In the field of oncology, FOXD3 is considered to be a tumor suppressor factor, which works by inhibiting cell proliferation, promoting apoptosis and activating MAPK / ERK signaling pathways. GBX2 is a transcriptional co-repressor that mainly inhibits gene expression by binding to transcription factors. It plays an important role in neurodevelopment and nervous system diseases, especially in neuronal differentiation and synaptic plasticity. SOX3 is an SRY homeobox transcription factor that participates in the regulation of a variety of biological processes, including development, differentiation and cell proliferation. It plays an important role in sex determination and reproductive system development during embryonic development. HIST1H4 is a variant of histone H4 that participates in the formation of chromatin structure and gene expression regulation. It plays an important role in maintaining genome stability and regulating gene expression.

[0004] There are no reports on the association of FOXD3, GBX2, SOX3 and HIST1H4 with doxorubicin-induced cardiotoxicity, especially with doxorubicin-induced cardiotoxicity in breast cancer chemotherapy.

[0005] SIRI is the Systemic Inflammation Response Index, a comprehensive indicator used to evaluate the systemic inflammatory state. It combines the ratio of neutrophils, monocytes and lymphocytes in the blood to evaluate the systemic inflammatory response. The calculation formula is:

[0006]

[0007] At present, there are no reports of combining FOXD3, GBX2, SOX3 and HIST1H4 with SIRI to assess the risk of doxorubicin-induced cardiotoxicity to avoid heart damage in patients during doxorubicin chemotherapy. Summary of the invention

[0008] The purpose of the present invention is to provide a group of new biomarkers for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer, and the use of detection reagents of these markers in the preparation of a kit for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer. By combining the expression of biomarkers in serum obtained by detection with the whole blood SIRI index, a prediction model for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer is constructed, so as to achieve accurate prediction of the occurrence of doxorubicin chemotherapy-induced cardiotoxicity in breast cancer.

[0009] The technical solution of the present invention includes:

[0010] Use of a reagent for detecting the expression level of a biomarker in the preparation of a kit for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer, characterized in that the biomarker is the FOXD3 gene and / or its expression product, the GBX2 gene and / or its expression product, the SOX3 gene and / or its expression product, or the HIST1H4 gene and / or its expression product.

[0011] Further, the reagent for detecting the expression level of FOXD3 gene includes a reagent for detecting FOXD3 mRNA or detecting FOXD3 protein;

[0012] The reagent for detecting the expression level of GBX2 gene includes a reagent for detecting GBX2 mRNA or detecting GBX2 protein;

[0013] The reagent for detecting the expression level of the SOX3 gene includes a reagent for detecting SOX3 mRNA or detecting SOX3 protein;

[0014] The reagent for detecting the expression level of HIST1H4 gene includes a reagent for detecting HIST1H4 mRNA or detecting HIST1H4 protein;

[0015] Furthermore, the reagents for detecting FOXD3 mRNA, GBX2 mRNA, SOX3 mRNA or HIST1H4 mRNA include reagents for RT-qPCR detection;

[0016] The RT-qPCR detection reagents include a primer pair for amplifying the FOXD3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 1 to 2; a primer pair for amplifying the GBX2 gene, whose nucleotide sequence is shown in SEQ ID NOs. 3 to 4; a primer pair for amplifying the SOX3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 5 to 6; or a primer pair for amplifying the HIST1H4 gene, whose nucleotide sequence is shown in SEQ ID NOs. 7 to 8;

[0017] The reagent for detecting FOXD3 protein, GBX2 protein, SOX3 protein or HIST1H4 protein includes an enzyme-linked immunosorbent assay reagent or an enzyme-linked immunosorbent assay reagent.

[0018] The present invention also provides a kit for predicting adriamycin chemotherapy-induced cardiac toxicity in breast cancer, which comprises reagents for detecting the expression level of biomarkers;

[0019] The biomarker is FOXD3 gene and / or its expression product, GBX2 gene and / or its expression product, SOX3 gene and / or its expression product, or HIST1H4 gene and / or its expression product;

[0020] The reagent for detecting the expression level of FOXD3 gene includes a reagent for detecting FOXD3 mRNA or detecting FOXD3 protein;

[0021] The reagent for detecting the expression level of GBX2 gene includes a reagent for detecting GBX2 mRNA or detecting GBX2 protein;

[0022] The reagent for detecting the expression level of the SOX3 gene includes a reagent for detecting SOX3 mRNA or detecting SOX3 protein;

[0023] The reagent for detecting the expression level of HIST1H4 gene includes a reagent for detecting HIST1H4 mRNA or detecting HIST1H4 protein;

[0024] The reagents for detecting FOXD3 mRNA, GBX2 mRNA, SOX3 mRNA or HIST1H4 mRNA include reagents for RT-qPCR detection;

[0025] The RT-qPCR detection reagents include a primer pair for amplifying the FOXD3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 1 to 2; a primer pair for amplifying the GBX2 gene, whose nucleotide sequence is shown in SEQ ID NOs. 3 to 4; a primer pair for amplifying the SOX3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 5 to 6; or a primer pair for amplifying the HIST1H4 gene, whose nucleotide sequence is shown in SEQ ID NOs. 7 to 8;

[0026] The reagent for detecting FOXD3 protein, GBX2 protein, SOX3 protein or HIST1H4 protein includes an enzyme-linked immunosorbent assay reagent or an enzyme-linked immunosorbent assay reagent.

[0027] The present invention also provides a use of the aforementioned screening kit in preparing a prediction model for predicting adriamycin chemotherapy-induced cardiac toxicity in breast cancer.

[0028] The present invention also provides a prediction model for predicting doxorubicin chemotherapy-induced cardiac toxicity in breast cancer, wherein the prediction model is a nomogram, which is composed of a score scale, a prediction variable axis, a total score scale, and a risk axis that are arranged in sequence from top to bottom and are parallel to each other;

[0029] The predictor variable axis consists of a FOXD3 predictor variable axis, a GBX2 predictor variable axis, a SOX3 predictor variable axis, a HIST1H4 predictor variable axis and a SIRI predictor variable axis;

[0030] The score scale is a scale that indicates the score corresponding to the scale on the prediction variable axis. The scale value of the scale is 0-100, with 0 at the leftmost end and 100 at the rightmost end. The scale of the scale is equally divided.

[0031] The FOXD3 predictor variable axis is a scale indicating the expression of FOXD3 protein in serum, the scale value of the scale is 50-400, 50 is at the far left end, 400 is at the far right end, and the scale of the scale is equally divided; when the scale is 50, the corresponding score on the score scale is 0, and when the scale is 400, the corresponding score on the score scale is 51;

[0032] The SOX3 prediction variable axis is a scale indicating the expression of SOX3 protein in serum, the scale value of the scale is 10-45, 10 is at the leftmost end, 45 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 10, the corresponding score on the score scale is 0, and when the scale is 45, the corresponding score on the score scale is 100;

[0033] The GBX2 predictor variable axis is a scale indicating the expression of GBX2 protein in serum, the scale value of the scale is 0-800, 0 is at the leftmost end, 800 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 0, the corresponding score on the score scale is 0, and when the scale is 800, the corresponding score on the score scale is 28;

[0034] The HIST1H4 prediction variable axis is a scale indicating the expression of HIST1H4 protein in serum, the scale value of the scale is 2-20, 2 is at the far left end, 20 is at the far right end, and the scale of the scale is equally divided; when the scale is 2, the corresponding score on the score scale is 0, and when the scale is 20, the corresponding score on the score scale is 48;

[0035] The SIRI predictor variable axis is a scale representing the whole blood SIRI index, the scale value of the scale is 0-7, 0 is at the leftmost end, 7 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 0, the corresponding score on the score scale is 0, and when the scale is 7, the corresponding score on the score scale is 78;

[0036] The total score scale is a scale that represents the sum of the scores corresponding to the scales on the prediction variable axis on the score scale. The scale value of the scale is 0-260, with 0 at the leftmost end and 260 at the rightmost end. The scale of the scale is equally divided; when the scale is 0, it corresponds to the 0 scale of the score scale, and when the scale is 260, it corresponds to the 100 scale of the score scale.

[0037] The risk axis is a scale representing the risk probability of doxorubicin-induced cardiotoxicity, the scale value of the scale is 0.01-0.99, 0.01 is at the leftmost end, 0.99 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 0.01, the corresponding score on the total score scale is 104, and when the scale is 0.99, the corresponding score on the total score scale is 150; risk probability = the value corresponding to the score on the total score scale on the risk axis, when the value corresponding to the score on the total score scale on the risk axis is at the left end of 0.01, the risk probability of doxorubicin-induced cardiotoxicity at this time is <0.01, and when the value corresponding to the score on the total score scale on the risk axis is at the right end of 0.99, the risk probability of doxorubicin-induced cardiotoxicity at this time is >0.99.

[0038] Furthermore, the prediction model is used to predict the probability of occurrence of cardiac toxicity caused by doxorubicin chemotherapy in breast cancer.

[0039] The present invention also provides a method for constructing the aforementioned prediction model, comprising the following steps:

[0040] (1) Collect prediction indicators and enter them into the input module;

[0041] (2) Use the indicators in the input module to build a logistic regression model, and use the RMS calculation package to complete the visualization of the nomogram of the logistic regression model;

[0042] The prediction indicators are the expression levels of FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in serum, and the whole blood SIRI index;

[0043] The expression level of the FOXD3 protein in serum is 50-400 pg / mL, the expression level of the GBX2 protein in serum is 0-800 ng / mL, the expression level of the HIST1H4 protein in serum is 2-20 ng / mL, the expression level of the SOX3 protein in serum is 10-45 ng / mL, and the whole blood SIRI index is 0-7.

[0044] The present invention also provides a use of the above-mentioned prediction model in preparing a device for predicting cardiac toxicity induced by adriamycin chemotherapy in breast cancer.

[0045] The present invention also provides a device for predicting doxorubicin-induced cardiac toxicity in breast cancer, wherein the device comprises the aforementioned prediction model.

[0046] The key to the present invention is that it is determined that the expression levels of FOXD3 gene, GBX2 gene, SOX3 gene and HIST1H4 gene in cells acted with doxorubicin are increased; the expression levels of FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in the serum of breast cancer patients undergoing doxorubicin chemotherapy are significantly correlated with doxorubicin-induced cardiotoxicity, so the expression levels of FOXD3, GBX2, SOX3 and HIST1H4 genes or their expression products can be used as a basis for predicting doxorubicin-induced cardiotoxicity. As for the means for specifically detecting the expression levels of FOXD3, GBX2, SOX3 and HIST1H4 genes or their expression products, various means disclosed in the prior art can be used. The embodiment of the present invention specifically uses RT-qPCR to detect FOXD3 gene, GBX2 gene, SOX3 gene and HIST1H4 gene in cells; ELISA is used to detect FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in serum, but it is not limited to these means. Any method that can detect the expression level of FOXD3 gene, GBX2 gene, SOX3 gene and HIST1H4 gene in cells, or the expression level of FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in serum can be used to obtain evaluation indicators for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer.

[0047] The key to the present invention is to construct a nomogram prediction model by combining the expression levels of FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in the serum with the whole blood SIRI index, so as to intuitively and accurately predict the occurrence of doxorubicin-induced cardiotoxicity, and provide conditions for early correction of cardiac function and avoidance of permanent heart damage in breast cancer patients who use doxorubicin for chemotherapy. The nomogram prediction model constructed based on the newly discovered biomarkers and SIRI of the present invention has high prediction accuracy and discrimination, and is of great significance for clinical evaluation of the occurrence of doxorubicin chemotherapy-induced cardiotoxicity in breast cancer, and is helpful for individual precision treatment of clinical patients.

[0048] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0049] The above contents of the present invention are further described in detail below through specific implementation methods. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 Verification results of cell expression levels of cardiotoxicity-related markers; (A) MDA-MB-231 cells, (B) AC16 cells;

[0051] Figure 2 Construction and validation of the prediction model; comparative analysis of serum expression of related proteins and SIRI index in breast cancer patients with and without cardiotoxicity caused by doxorubicin chemotherapy (A), deviation analysis (B), coefficient path diagram (C) and ROC curve diagram (D); nomogram model based on five factors (E), ROC curve diagram (F), precision-recall PR curve diagram (G), decision curve diagram (H), clinical impact curve diagram (I) and calibration curve diagram (J). DETAILED DESCRIPTION

[0052] Example 1 Study on biomarkers in serum associated with cardiotoxicity induced by adriamycin chemotherapy in breast cancer

[0053] 1. Bioinformatics screening

[0054] The biological information of breast cancer patients who used doxorubicin chemotherapy and those who did not was analyzed using TCGA / GEO database, WGCNA method, prognostic analysis and other analysis tools.

[0055] Bioinformatics results showed that compared with the group not using doxorubicin chemotherapy, the mRNA levels of FOXD3, GBX2, SOX3 and HIST1H4 were increased in breast cancer patients using doxorubicin chemotherapy. At the same time, the high expression of these genes was negatively correlated with the prognosis of breast cancer patients. The FOXD3, GBX2, SOX3 and HIST1H4 genes or the proteins encoded by the genes were initially screened as biomarkers for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer.

[0056] 2. Verification of the cellular expression level of markers related to cardiotoxicity induced by doxorubicin chemotherapy in breast cancer

[0057] 2.1 Methods

[0058] For verification, breast cancer cells MDA-MB-231 and cardiomyocytes AC16 were selected, and cells were treated with 2μM concentration of doxorubicin. After the cells were cultured for the same time, the cells were collected, and the cell RNA was extracted, and then reverse transcribed into cDNA, and then qPCR detection was performed. At the same time, breast cancer cells MDA-MB-231 and cardiomyocytes AC16 that were not treated with doxorubicin were used as controls. RT-qPCR detection was performed as follows:

[0059] 1) RNA reverse transcription

[0060] TAKARA RT-Mastermix 4ul

[0061] Total RNA 2000ng

[0062] Make up to 20ul with RNase-ffee water

[0063] Reaction conditions: 37°C, 15 min; 85°C, 5 s; stored at 4°C.

[0064] 2) PCR reaction

[0065] TAKARA SYBR Green 5ul

[0066] Primer mix (10M) 0.8ul

[0067] cDNA 1ul

[0068] DNase-free water 3.2ul

[0069] Reaction conditions: 95°C, 30 s, 1 cycle; 95°C, 5 s, 60°C, 30 s, 40 cycles; melting curve: 65.0°C to 95°C, temperature gradient of 0.5°C, time of 5 s; Real-Time PCR analysis was performed using a fluorescent quantitative PCR instrument from Bio-Rad, USA.

[0070] Primer sequences:

[0071]

[0072] 2.2 Verification Results

[0073] The expression levels of FOXD3, GBX2, SOX3 and HIST1H4 genes in MDA-MB-231 and AC16 cells were increased after treatment with doxorubicin, and the difference was statistically significant ( Figure 1 ), indicating that the cardiotoxicity caused by doxorubicin chemotherapy in breast cancer can be evaluated by detecting the expression levels of FOXD3, GBX2, SOX3 and HIST1H4 genes in breast cancer cells or cardiomyocytes.

[0074] 3. Clinical validation of markers related to cardiotoxicity induced by doxorubicin chemotherapy in breast cancer

[0075] 1) Clinical Cases

[0076] A total of 80 breast cancer patients who received doxorubicin chemotherapy in the Affiliated Hospital of Southwest Medical University were selected for validation. Among them, 35 breast cancer patients with cardiotoxicity caused by doxorubicin chemotherapy were selected as the experimental group, and 45 breast cancer patients with no cardiotoxicity caused by doxorubicin chemotherapy were selected as the control group. There was no significant difference in general data between the two groups (P>0.05), and the two groups were comparable.

[0077] 2) Serological testing

[0078] ELISA kits were used to detect the expression levels of FOXD3, GBX2, SOX3 and HIST1H4 in serum of patients in the experimental group and the control group.

[0079] 3) Whole blood SIRI index test

[0080] The whole blood cells were counted using a blood cell counter. The patient's SIRI was calculated using the whole blood routine test results = monocyte count × neutrophil count / lymphocyte count.

[0081] 4) Build a prediction model

[0082] LASSO regression (least absolute shrinkage and selection operator regression) method was used for feature selection. LASSO regression screened out clinical markers closely related to adverse cardiac events (including FOXD3, GBX2, SOX3, HIST1H4 and SIRI) and constructed a prediction model. The ROC curve was used to evaluate the AUC value of each factor to display the variables with predictive ability. Based on the LASSO regression results, a nomogram model containing five factors was constructed using the Logistic regression model, in which the RMS calculation package was used to complete the nomogram visualization. ROC curve and precision-recall (PR) curve analysis were used to verify the predictive performance of the model. Calibration curve and decision curve analysis (DCA) were used to evaluate the clinical applicability and predictive value of the model.

[0083] The results of ELISA analysis of clinical samples showed that the levels of FOXD3, GBX2, SOX3 and HIST1H4 were significantly increased in the serum of breast cancer patients who developed cardiotoxicity after receiving doxorubicin chemotherapy ( Figure 2 A). At the same time, the ROC curve analysis of the SIRI calculated from the patient's serum whole blood data and the diagnostic model constructed by combining the four factors and SIRI showed that the AUCs of FOXD3, GBX2, SOX3, HIST1H4 and SIRI were 0.743, 0.724, 0.893, 0.760, and 0.776, respectively ( Figure 2 B~D).

[0084] The nomogram model based on five factors ( Figure 2 E) Display:

[0085] The nomogram is composed of a score scale, a predictor variable axis, a total score scale, and a risk axis that are arranged in order from top to bottom and are parallel to each other, wherein the predictor variable axis is composed of the FOXD3 predictor variable axis, the SOX3 predictor variable axis, the GBX2 predictor variable axis, the HIST1H4 predictor variable axis, and the SIRI predictor variable axis;

[0086] The score scale is a scale that represents the score corresponding to the scale on the predictor variable axis. The scale value of the scale is 0-100, with 0 at the far left and 100 at the far right. The scale of the scale is equally divided;

[0087] The FOXD3 predictor variable axis is a scale indicating the expression of FOXD3 protein in serum. The scale value of the scale is 50-400, with 50 at the far left and 400 at the far right. The scale of the scale is equally divided. When the scale is 50, the corresponding score on the score scale is 0, and when the scale is 400, the corresponding score on the score scale is 51.

[0088] The SOX3 predictor variable axis is a scale indicating the expression of SOX3 protein in serum. The scale value of the scale is 10-45, with 10 at the far left and 45 at the far right. The scale of the scale is equally divided. When the scale is 10, the corresponding score on the score scale is 0, and when the scale is 45, the corresponding score on the score scale is 100.

[0089] The GBX2 predictor variable axis is a scale indicating the expression of GBX2 protein in serum. The scale value of the scale is 0-800, with 0 at the far left and 800 at the far right. The scale is divided equally. When the scale is 0, the corresponding score on the score scale is 0, and when the scale is 800, the corresponding score on the score scale is 28.

[0090] The HIST1H4 predictor variable axis is a scale indicating the expression of HIST1H4 protein in serum. The scale value of the scale is 2-20, with 2 at the far left and 20 at the far right. The scale of the scale is equally divided. When the scale is 2, the corresponding score on the score scale is 0, and when the scale is 20, the corresponding score on the score scale is 48.

[0091] The SIRI predictor variable axis is a scale representing the whole blood SIRI index. The scale value of the scale is 0-7, with 0 at the far left and 7 at the far right. The scale of the scale is equally divided. When the scale is 0, the corresponding score on the score scale is 0, and when the scale is 7, the corresponding score on the score scale is 78.

[0092] The total score scale is a scale that represents the sum of the scores corresponding to the scales on the prediction variable axis on the score scale. The scale value is 0-260, with 0 at the far left and 260 at the far right. The scale is equally divided; when the scale is 0, it corresponds to the 0 scale of the score scale, and when the scale is 260, it corresponds to the 100 scale of the score scale.

[0093] The risk axis is a scale representing the risk probability of doxorubicin-induced cardiotoxicity. The scale value of the scale is 0.01-0.99, with 0.01 at the far left and 0.99 at the far right. The scale of the scale is equally divided; When the scale is 0.01, the corresponding score on the total score scale is 104, and when the scale is 0.99, the corresponding score on the total score scale is 150; risk probability = the value on the risk axis corresponding to the score on the total score scale. When the value on the risk axis corresponding to the score on the total score scale is at the left end of 0.01, the risk probability of doxorubicin-induced cardiotoxicity is <0.01; when the value on the risk axis corresponding to the score on the total score scale is at the right end of 0.99, the risk probability of doxorubicin-induced cardiotoxicity is >0.99.

[0094] The validation results of the predictive performance of the nomogram model: the AUC of the ROC curve was 0.963 ( Figure 2 F), the AUC of the precision recall PR curve is 0.973 ( Figure 2 G), and the decision curve ( Figure 2 H), clinical impact curve ( Figure 2 I) and calibration curve ( Figure 2 J) showed that the nomogram model containing five factors has good predictive performance and clinical application value. In addition, the clinical calibration curve of the model is Dxy = 0.926, indicating that the nomogram model has good discrimination ability and can accurately predict the cardiac toxicity induced by doxorubicin chemotherapy in breast cancer.

[0095] In summary, the present invention uses bioinformatics methods to screen the expression levels of FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in serum, which are significantly correlated with doxorubicin-induced cardiotoxicity. These newly discovered biomarkers are combined with SIRI as predictive indicators, and a nomogram is constructed through a Logistic regression model. The obtained nomogram prediction model is verified by ROC curve, PR curve, policy curve, clinical impact curve and calibration curve in many aspects to determine that its prediction accuracy and discrimination are high, which is of great significance for clinical evaluation of the occurrence of doxorubicin chemotherapy-induced cardiotoxicity in breast cancer, and is helpful for individual precision treatment of clinical patients.

Claims

1. Use of a reagent for detecting the expression level of a biomarker in the preparation of a kit for predicting adriamycin chemotherapy-induced cardiotoxicity in breast cancer, characterized in that: The biomarker is FOXD3 gene and / or its expression product, GBX2 gene and / or its expression product, SOX3 gene and / or its expression product, or HIST1H4 gene and / or its expression product.

2. The use according to claim 1, characterized in that: The reagent for detecting the expression level of FOXD3 gene includes a reagent for detecting FOXD3 mRNA or detecting FOXD3 protein; The reagent for detecting the expression level of GBX2 gene includes a reagent for detecting GBX2 mRNA or detecting GBX2 protein; The reagent for detecting the expression level of the SOX3 gene includes a reagent for detecting SOX3 mRNA or detecting SOX3 protein; The reagent for detecting the expression level of HIST1H4 gene includes a reagent for detecting HIST1H4 mRNA or a reagent for detecting HIST1H4 protein.

3. The use according to claim 2, characterized in that The reagents for detecting FOXD3 mRNA, GBX2 mRNA, SOX3 mRNA or HIST1H4 mRNA include reagents for RT-qPCR detection; The RT-qPCR detection reagents include a primer pair for amplifying the FOXD3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 1 to 2; a primer pair for amplifying the GBX2 gene, whose nucleotide sequence is shown in SEQ ID NOs. 3 to 4; a primer pair for amplifying the SOX3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 5 to 6; or a primer pair for amplifying the HIST1H4 gene, whose nucleotide sequence is shown in SEQ ID NOs. 7 to 8; The reagent for detecting FOXD3 protein, GBX2 protein, SOX3 protein or HIST1H4 protein includes an enzyme-linked immunosorbent assay reagent or an enzyme-linked immunosorbent assay reagent.

4. A kit for predicting adriamycin chemotherapy-induced cardiotoxicity in breast cancer, characterized in that: It includes reagents for detecting the expression level of the biomarker; The biomarker is FOXD3 gene and / or its expression product, GBX2 gene and / or its expression product, SOX3 gene and / or its expression product, or HIST1H4 gene and / or its expression product; The reagent for detecting the expression level of FOXD3 gene includes a reagent for detecting FOXD3 mRNA or detecting FOXD3 protein; The reagent for detecting the expression level of GBX2 gene includes a reagent for detecting GBX2 mRNA or detecting GBX2 protein; The reagent for detecting the expression level of the SOX3 gene includes a reagent for detecting SOX3 mRNA or detecting SOX3 protein; The reagent for detecting the expression level of HIST1H4 gene includes a reagent for detecting HIST1H4 mRNA or detecting HIST1H4 protein; The reagents for detecting FOXD3 mRNA, GBX2 mRNA, SOX3 mRNA or HIST1H4 mRNA include reagents for RT-qPCR detection; The RT-qPCR detection reagents include a primer pair for amplifying the FOXD3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 1 to 2; a primer pair for amplifying the GBX2 gene, whose nucleotide sequence is shown in SEQ ID NOs. 3 to 4; a primer pair for amplifying the SOX3 gene, whose nucleotide sequence is shown in SEQ ID NOs. 5 to 6; or a primer pair for amplifying the HIST1H4 gene, whose nucleotide sequence is shown in SEQ ID NOs. 7 to 8; The reagent for detecting FOXD3 protein, GBX2 protein, SOX3 protein or HIST1H4 protein includes an enzyme-linked immunosorbent assay reagent or an enzyme-linked immunosorbent assay reagent.

5. Use of the kit according to claim 4 in preparing a prediction model for predicting adriamycin chemotherapy-induced cardiotoxicity in breast cancer.

6. A prediction model for predicting adriamycin chemotherapy-induced cardiotoxicity in breast cancer, characterized in that: The prediction model is a nomogram, which is composed of a score scale, a prediction variable axis, a total score scale and a risk axis arranged in sequence from top to bottom and parallel to each other; The predictor variable axis consists of a FOXD3 predictor variable axis, a GBX2 predictor variable axis, a SOX3 predictor variable axis, a HIST1H4 predictor variable axis and a SIRI predictor variable axis; The score scale is a scale that indicates the score corresponding to the scale on the prediction variable axis. The scale value of the scale is 0-100, with 0 at the leftmost end and 100 at the rightmost end. The scale of the scale is equally divided. The FOXD3 predictor variable axis is a scale indicating the expression of FOXD3 protein in serum, the scale value of the scale is 50-400, 50 is at the far left end, 400 is at the far right end, and the scale of the scale is equally divided; when the scale is 50, the corresponding score on the score scale is 0, and when the scale is 400, the corresponding score on the score scale is 51; The SOX3 prediction variable axis is a scale indicating the expression of SOX3 protein in serum, the scale value of the scale is 10-45, 10 is at the leftmost end, 45 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 10, the corresponding score on the score scale is 0, and when the scale is 45, the corresponding score on the score scale is 100; The GBX2 predictor variable axis is a scale indicating the expression of GBX2 protein in serum, the scale value of the scale is 0-800, 0 is at the leftmost end, 800 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 0, the corresponding score on the score scale is 0, and when the scale is 800, the corresponding score on the score scale is 28; The HIST1H4 prediction variable axis is a scale indicating the expression of HIST1H4 protein in serum, the scale value of the scale is 2-20, 2 is at the far left end, 20 is at the far right end, and the scale of the scale is equally divided; when the scale is 2, the corresponding score on the score scale is 0, and when the scale is 20, the corresponding score on the score scale is 48; The SIRI predictor variable axis is a scale representing the whole blood SIRI index, the scale value of the scale is 0-7, 0 is at the leftmost end, 7 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 0, the corresponding score on the score scale is 0, and when the scale is 7, the corresponding score on the score scale is 78; The total score scale is a scale that represents the sum of the scores corresponding to the scales on the prediction variable axis on the score scale. The scale value of the scale is 0-260, with 0 at the leftmost end and 260 at the rightmost end. The scale of the scale is equally divided; when the scale is 0, it corresponds to the 0 scale of the score scale, and when the scale is 260, it corresponds to the 100 scale of the score scale. The risk axis is a scale representing the risk probability of doxorubicin-induced cardiotoxicity, the scale values ​​of the scale are 0.01-0.99, 0.01 is at the leftmost end, 0.99 is at the rightmost end, and the scale of the scale is equally divided; when the scale is 0.01, the corresponding score on the total score scale is 104, and when the scale is 0.99, the corresponding score on the total score scale is 150; risk probability = the value corresponding to the score on the total score scale on the risk axis, when the value corresponding to the score on the total score scale on the risk axis is at the left end of 0.01, the risk probability of doxorubicin-induced cardiotoxicity at this time is <0.01, and when the value corresponding to the score on the total score scale on the risk axis is at the right end of 0.99, the risk probability of doxorubicin-induced cardiotoxicity at this time is >0.

99.

7. The prediction model according to claim 6, wherein the prediction model is used to predict the probability of occurrence of cardiac toxicity caused by doxorubicin chemotherapy for breast cancer.

8. A method for constructing a prediction model according to claim 6 or 7, characterized in that: The steps include: (1) Collect prediction indicators and enter them into the input module; (2) Use the indicators in the input module to build a logistic regression model, and use the RMS calculation package to complete the visualization of the nomogram of the logistic regression model; The prediction indicators are the expression levels of FOXD3 protein, GBX2 protein, SOX3 protein and HIST1H4 protein in serum, and the whole blood SIRI index; The expression level of the FOXD3 protein in serum is 50-400 pg / mL, the expression level of the GBX2 protein in serum is 0-800 ng / mL, the expression level of the HIST1H4 protein in serum is 2-20 ng / mL, the expression level of the SOX3 protein in serum is 10-45 ng / mL, and the whole blood SIRI index is 0-11.

9. Use of the prediction model according to any one of claims 6 or 7 in the preparation of a device for predicting doxorubicin chemotherapy-induced cardiotoxicity in breast cancer.

10. A device for predicting cardiac toxicity induced by adriamycin chemotherapy in breast cancer, characterized in that: The device comprises the prediction model according to claim 6 or 7.

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