Multilevel Prediction-mediated Prognosis Of Breast Cancer And Comprehensive Evaluation Of Cardiac Toxicity Of Chemotherapy

Background:Breast cancer is one of the most common cancers among women in the world.Complex immune processes are involved in the occurrence and progression of breast cancer.There is abundant evidence that biomarkers play a key role in predicting the prognosis of breast cancer.In this case,a variety of novel biomarkers,such as proteins and non-coding RNA,have been studied,which provides a new therapeutic idea for clinic.However,known single genes can not accurately predict D in breast cancer patients or be used in clinical practice.At the same time,the combination of biomarkers can improve the sensitivity and specificity of prognostic D measurement in breast cancer patients.Therefore,it is necessary to establish an expression-based gene marker to predict the overall survival rate（OS）of patients with breast cancer,which is essential for the clinical decision-making of the best treatment plan.Cancer monotherapy generally has some disadvantages that are difficult to overcome,such as limited treatment benefit and strong systemic toxicity.Combination therapy is generally considered to be the preferred treatment for cancer.Epidermal growth factor receptor has been used in many breast cancers.Expression,targeted drugs（trastuzumab,cetuximab,etc.）targeting epidermal growth factor receptor（EGFR）,have a therapeutic effect in reducing breast cancer recurrence,so how to improve the resistance of targeted drugs Cancer effects have also become a new hot spot.However,the cardiotoxicity of the targeted drug itself is one of the most devastating complications,especially in the combination of anthracyclines,the mortality of heart failure caused by anthracycline treatment is significantly increased^[1].Cardiotoxicity has become a major threat to the survival of cancer patients.Therefore,it is important to determine the risk of left ventricular dysfunction in patients at an early stage.These patients may benefit from the adjustment of their cancer treatment regimen and the initiation of cardioprotective measures^[2].Therefore,predicting the prognosis of breast cancer patients through genetic maps may be a better choice,and it is imperative to predict the toxicity of patients’hearts during treatment through various indicators.In order to achieve the maximum synergistic effect of the combination therapy,it is necessary to use the same carrier to simultaneously deliver the exact doses of the two preparations to the same cancer cell,so that the two drugs can exert synergistic effects to achieve a desired anticancer effect.Objective:There is ample evidence that biomarkers play a key role in predicting breast cancer prognosis.However,a single prognostic analysis of biomarkers is not sufficient,suggesting that genetic maps may be a better choice.Our goal is to establish a more accurate immune-related genetic map to predict the prognosis of breast cancer patients,and to evaluate the use of two-dimensional speckle tracking technology and blood biochemical markers in breast cancer patients-high sensitivity cardiac troponin I（High-sensitivity troponinI,hsTNI）,Heart-type fatty acid-binding protein（H-FABP）,N-terminal pro-B-type natriuretic peptide,NT-proBNP)Can predict the development of early cardiotoxicity.At the same time,this study also combined silicon nanoparticles with cetuximab（Cet-SLN）,targeting epidermal growth factor receptor（EGFR）,and verifying whether Cet-SLN/ICG can be used as breast cancer chemotherapy-photothermal Whether the potential carrier of the combination therapy has a stronger anticancer effect.Method:Information on 1,203 breast cancer patients was obtained from The Cancer Genome Atlas（TCGA）.Gene set enrichment analysis（GSEA）confirmed that antigen processing presented gene sets are of significance in breast cancer.The Cox proportional regression model was used to identify three genes（HSPA5（Hsp70 family member 5）,PSME2（proteasome activator subunit 2）,HLA-F（HSPA-F）Major histocompatibility complex,class I,F）).Multivariate Cox regression and stratified analysis showed that the prognostic power of the three gene markers was not related to other clinical variables.102 patients with pathologically confirmed breast cancer were HER-2 positive and received A（E）C（doxorubicin or Epirubicin,cyclophosphamide）for 4 cycles,followed by paclitaxel plus trastuzumab.Clinical routine echocardiographic parameters and overall longitudinal strain were measured at baseline,3 months after chemotherapy（3weeks after the last anthracycline cycle）,6 months after chemotherapy（3 months after trastuzumab treatment）.GLS),circumferential contraction strain（GCS）,N-terminal pro-B-type natriuretic peptide（NT-proBNP）and high-sensitivity cardiac troponin I（hsTNI）.At the same time,before the chemotherapy and 24 hours after the administration,blood samples were taken at different time points for H-FABP and high-sensitivity troponin I（hsTNI）measurement,and parallel echocardiography was performed to obtain myocardial strain imaging-total longitudinal strain（GLS）and circumferential contraction strain（GCS）.Patients were divided into a cardiotoxic group and a non-cardiotoxic group according to diagnostic criteria for cardiotoxicity.To evaluate the predictive value of H-FABP,hsTNI,NT-proBNP,GLS and GCS for cardiotoxicity induced by chemotherapeutic drugs.In addition,we use Cet-SLN’s high drug loading capacity to encapsulate the photothermotherapy reagent phthalocyanine green（ICG）,which ultimately results in a multi-purpose drug delivery system（DDS）that delivers both Cet and ICG（Cet-SLN/ICG）for the combined treatment of breast cancer chemotherapy-photothermal.Result:The three-gene map predicts the prognosis of the patient,and the risk score generated by the map may be an independent prognostic indicator.At the same time,within 24 hours after the first infusion of chemotherapeutic drugs（adriamycin or Epirubicin,cyclophosphamide）,there was no significant difference in the level of H-FABP and GLS between non-cardiotoxic group and non-cardiotoxic group.High sensitivity cardiac troponin I was significantly different from baseline 24 hours after chemotherapy.Before the start of trastuzumab therapy（3 months after chemotherapy）,GLS in cardiotoxic group decreased from-20.30（+1.56%）at baseline to-15.30（+2.98%）.Compared with non-cardiotoxic group,GLS in cardiotoxic group decreased significantly at 3 months after treatment（P<0.05）,while the lowering of left ventricular ejection fraction was most common within 6 months after treatment,and less common within 3 months after treatment.The mean left ventricular ejection fraction decreased from 62.90（+2.2）to 52.40（+2.98%）within 6 months,which was significantly different from 60.21（+2.58%）in normal patients（P<0.05）.At 6 months,the average GCS value of the cardiotoxic group was-16.57+2.07%,which was significantly different from that of the non-cardiotoxic group（P<0.05）.High sensitivity cardiac troponin increased significantly at 3 months,which was significantly different from the control group（P<0.05）.The diagnostic thresholds of hsTnI and GLS obtained from ROC curve analysis were used as the dividing lines（1007 ug/L,-17.5%）.According to hsTnI and GLS were all greater than the thresholds（two high groups）and one of them was greater than the thresholds（one high group）,the values were assigned to 1 and 0 respectively.The binary logistic analysis was carried out after defining the new variables.From the results,the predictive probability was 0.976.In patients treated with anthracyclines and trastuzumab,GLS and high sensitivity cardiac troponin I（hsTNI）are easy to measure,can identify subtle changes in the heart,and can predict subsequent decrease in left ventricular ejection fraction.NT-pro-BNP increased significantly at 6 months compared with the control group（P<0.05）.The level of H-FABP did not increase significantly and was not helpful in predicting early cardiac dysfunction associated with anthracycline and trastuzumab.The results suggest that Cet-SLN/ICG is a kind of nanoparticle with good dispersibility and high stability under physiological conditions.Due to the combination of cetuximab,Cet-SLN/ICG can target epidermal growth factor receptors that are overexpressed in MCF-7 cells,increasing their tumor targeting ability and cellular uptake effectiveness.Upon internalization by cancer cells,disulfide bonds can react with glutathione in the cytoplasm,triggering the rapid release of cetuximab and indocyanine green.After contact with near-infrared light,the two drugs can exert synergistic effects to achieve an ideal anti-cancer effect.Conclusion:We have established a three-gene map that can be used as a novel independent biomarker for predicting OS in breast cancer patients,and has certain reference value for the clinical treatment of breast cancer patients.GLS and high-sensitivity myocardial troponin levels predict changes in preclinical left ventricular systolic function and can predict subsequent EF reduction in breast cancer patients undergoing chemotherapy and targeted therapy.A decrease in GLS from baseline to 3 months and an increase in hsTNI levels at 3 months were predictors of cardiotoxicity development at 6 months of treatment.The combination of hsTNI levels and GLS can accurately predict the development of cardiotoxicity in chemotherapy and targeted patients.These two parameters may be helpful in detecting patients with possible cardiotoxicity and adjusting the chemotherapy regimen or replacement therapy in a timely manner,which may reduce the incidence of cardiotoxicity and its associated morbidity and mortality.The N-terminal pro-B-type natriuretic peptide can reflect cardiotoxicity,but is not sensitive enough in the early stage.H-FABP as an indicator of predicting cardiotoxicity needs further study.Cet-SLN/ICG can be used as a potential carrier for the combination of chemotherapy and photothermal therapy for breast cancer.Both in vitro and in vivo studies have shown that Cet-SLN/ICG has Cet-SLN/ICG compared to cetuximab or indocyanine green alone.Stronger anti-cancer effect.More importantly,both in vitro and in vivo studies have shown that Cet-SLN/ICG has a stronger anticancer effect than cetuximab or indocyanine green alone.