Cuproptosis基因和特征基因预测急性髓系白血病预后、免疫和肿瘤微环境的价值

目的筛选急性髓系白血病(AML)中的差异预后Cuproptosis基因和特征基因并探索其在AML中的预后情况以及在免疫和肿瘤微环境中的生物学作用和相关性。方法利用从TCGA、GEO和UCSC 3个主要数据库下载的AML临床、转录组、基因组和拷贝数数据,并从已发表的研究中收集Cuproptosis基因,从多组学的角度,通过各种生物信息学方法、Meta分析和二次分型,研究Cuproptosis基因和特征基因在生存、免疫、肿瘤微环境、干细胞相关性和药物敏感性等方面作用。结果1个Cuproptosis基因被鉴定为AML的差异预后基因和5个通过影响Cuproptosis进而影响AML患者预后的特征基因,并建立了预后模型。差异基因主要富集于线粒体活性、氧化还原酶和能量代谢。在免疫方面,巨噬细胞M0、中性粒细胞、活化记忆CD4 T细胞和Tregs与风险评分呈正相关,巨噬细胞M2、静息肥大细胞、辅助滤泡T细胞和记忆B细胞与风险评分呈负相关;在肿瘤微环境方面,低危组的免疫细胞评分低于高危组,并且在总评分中,低危组的肿瘤微环境得分同样低于高危组,说明高危组的肿瘤纯度低于低危组;然而,干细胞在高风险和低风险组之间没有显著相关性,共有14种药物对治疗AML敏感。结论通过构建AML预后模型,显示AML中的差异预后Cuproptosis基因和特征基因与免疫和肿瘤微环境密切相关。

Copper Metabolism and Cuproptosis:Molecular Mechanisms and Therapeutic Perspectives in Neurodegenerative Diseases

Copper is an essential trace element,and plays a vital role in numerous physiological processes within the human body.During normal metabolism,the human body maintains copper homeostasis.Copper deficiency or excess can adversely affect cellular function.Therefore,copper homeostasis is stringently regulated.Recent studies suggest that copper can trigger a specific form of cell death,namely,cuproptosis,which is triggered by excessive levels of intracellular copper.Cuproptosis induces the aggregation of mitochondrial lipoylated proteins,and the loss of iron-sulfur cluster proteins.In neurodegenerative diseases,the pathogenesis and progression of neurological disorders are linked to copper homeostasis.This review summarizes the advances in copper homeostasis and cuproptosis in the nervous system and neurodegenerative diseases.This offers research perspectives that provide new insights into the targeted treatment of neurodegenerative diseases based on cuproptosis.

ZNT1 Involves Cuproptosis through Regulating MTF1-conduced Expression of MT1X under Copper Overload

Industrial activities such as smelting emissions,mineral combustion and industrial wastewater discharge might lead to copper pollution in the environment.This kind of copper pollution has harmful effects on aquatic o rganisms,plants and animals through direct or indirect exposure.However,the current understanding of the toxicity of copper is rather limited.Copper overload can perturb intracellular homeostasis and induce oxidative stress and e ven cell death.Recently,cuproptosis has been identified as a copper-dependent form of cell death induced by o xidative stress in mitochondria.We uncover here that zinc transporter 1(ZNT1)is an important regulator involved in cuproptosis.Firstly,we established the copper overload-induced cell death model with the overexpression of copper importer SLC31A1 in HeLa cells.Using this model,we conducted unbiased genome-wide CRISPR-Cas9 screens in cells treated with copper.Our results revealed a significant enrichment of ZNT1 gene in both library A and library B plasmids.Knocking out of ZNT1 in HeLa cells notably prevented cuproptosis.Subsequent knockout of metal transcription factor 1(MTF1)in ZNT1-deficient cells nearly abolished their ability to resist copper-induced cell death.However,overexpression of metallothionein 1X(MT1X)in the double-knockout cells could p artially restored the resistance to cuproptosis by loss of MTF1.Mechanistically,knockout of ZNT1 could promote MT1X expression by activating MTF1.As a consequence,the interaction between MT1X and copper was e nhanced,reducing the flow of copper into mitochondria and eliminating mitochondria damage.Taken together,this study reveals the important role of ZNT1 in cuproptosis and shows MTF1-MT1X axis mediated resistance to c uproptosis.Moreover,our study will help to understand the regulatory mechanism of cellular and systemic copper homeostasis under copper overload,and present insights into novel treatments for damages caused by both genetic copper overload diseases and environmental copper contamination.

Effect of cuproptosis on acute kidney injury after cardiopulmonary bypass in diabetic patients

BACKGROUND Cardiopulmonary bypass(CPB)is a common procedure in cardiac surgery.CPB is a high-risk factor for acute kidney injury(AKI),and diabetes is also such a factor.Diabetes can lead to copper overload.It is currently unclear whether AKI after CPB in diabetic patients is related to copper overload.AIM To explore whether the occurrence of CPB-AKI in diabetic patients is associated with cuproptosis.METHODS Blood and urine were collected from clinical diabetic and non-diabetic patients before and after CPB.Levels of copper ion,lactate,glucose,heat shock protein-70(HSP-70),and dihydrolipoamide dehydrogenase(DLAT)were determined.A diabetic rat model was established and CPB was performed.The rats were assessed for the development of CPB-AKI,and for the association of AKI with cuproptosis by detecting copper levels,iron-sulfur cluster proteins and observation of mitochondrial structure by electron microscopy.RESULTS CPB resulted in elevations of copper,lactate,HSP-70 and DLAT in blood and urine in both diabetic and nondiabetic patients.CPB was associated with pathologic and mitochondrial damage in the kidneys of diabetic rats.Cuproptosis-related proteins also appeared to be significantly reduced.CONCLUSION CPB-AKI is associated with cuproptosis.Diabetes mellitus is an important factor aggravating CPB-AKI and cuproptosis.

Prediction of prognosis,immune escape and drug sensitivity of lung adenocarcinoma based on Cuproptosis-related LncRNA

Background:Lung Adenocarcinoma(LUAD)is the leading cause of death from lung cancer.Cuproptosis is the latest discovered way of programmed cell death,and Cuproptosis-Related Gene(CRG)is associated with the risk of LUAD.At present,there are few research of LUAD and Cuproptosis focuses on Long non-coding RNA(LncRNA).As genomics advances,LncRNA emerges as a potential target for understanding tumor progression and prognosis,offering prospects for biological targeted therapy.Therefore,this study provides new biomarkers and therapeutic targets for LUAD from the perspective of LncRNA.Methods:Gene expression,clinical outcome and gene mutation data of LUAD patients were downloaded from TCGA database.Spearman correlation was used to analyze the correlation between LncRNA and CRG.Univariate Cox,multivariate Cox and LASSO Cox regression analysis were used to construct a prognostic model of Cuproptosis-LncRNAs.GO and KEGG enrichment and immune function analysis were performed on differentially expressed genes between different risk groups.Then,immune escape analysis was performed on LUAD patients with different TIDE score.Finally,drug sensitivity analysis was performed on these differentially expressed genes.Results:A total of 2244 Cuproptosis-LncRNAs were found.Through the application of univariate Cox regression analysis,multivariate Cox regression analysis,and LASSO Cox regression analysis,a prognostic model was developed,integrating 15 Cuproptosis-LncRNAs to assess the risk of mortality.Following that,the model underwent assessment through risk score analysis,Kaplan-Meier survival analysis,risk distribution,and evaluation of survival outcomes.The results revealed an AUC value of 0.755 for the model,surpassing the AUC of other clinical pathological features.The results of KEGG analysis showed that the differentially expressed genes in different model groups were mainly involved in Amoebiasis,Fat digestion and absorption,and other signaling pathways.The results of TMB showed that the prognostic model of TMB combined with risk score could well evaluate the prognosis of patients.The TIDE scores did not exhibit a notable distinction between the two risk models.Analysis of drug sensitivity revealed that individuals in the low-risk category demonstrated greater responsiveness to 5-Fluorouracil,Axitinib,Bexarotene,and other drugs compared to those in the high-risk group.Conclusion:Our research offers a valuable reference for predicting the prognosis of LUAD,contributing to a better understanding of the future elucidation of the process and mechanism of Cuproptosis-LncRNAs in LUAD.

Advances in Cuproptosis of Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) has already become a severe health risk and brings a lot of healthcare burden to the world. Apart from traditional HCC treatment strategies (surgery, liver transplantation, etc.), the emergence of immunotherapy targeting the immune microenvironment of HCC has brought new promise to patients with advanced HCC. However, adverse effects like drug resistance still exist. The liver is the main organ for storing copper ions, in copper overload can lead to liver function impairment and even the development of HCC. In recent years, a new mode of cell death has been identified, namely cuproptosis, a mode of programmed cell death that is dependent on copper ions and the tricarboxylic acid (TCA) cycle with mitochondria. Interestingly, a potential relationship between cuproptosis and the development of HCC has been found. Conclusively, this review provides an in-depth discussion of copper homeostasis in humans, the mechanism of cuproptosis, the potential impact of cuproptosis with HCC, and the therapeutic modalities of HCC that target cuproptosis, which provide new insights to promote the development of research targeting cuproptosis in HCC.

Progress in the research of cuproptosis and possible targets for cancer therapy

Developing novel cancer therapies that exploit programmed cell death pathways holds promise for advancing cancer treatment.According to a recently published study in Science,copper death(cuproptosis)occurs when intracellular copper is overloaded,triggering aggregation of lipidated mitochondrial proteins and Fe–S cluster proteins.This intriguing phenomenon is triggered by the instability of copper ions.Understanding the molecular mechanisms behind cuproptosis and its associated genes,as identified by Tsvetkov,including ferredoxin 1,lipoic acid synthase,lipoyltransferase 1,dihydrolipid amide dehydrogenase,dihydrolipoamide transacetylase,pyruvate dehydrogenaseα1,pyruvate dehydrogenaseβ,metallothionein,glutaminase,and cyclin-dependent kinase inhibitor 2A,may open new avenues for cancer therapy.Here,we provide a new understanding of the role of copper death and related genes in cancer.

Drug repositioning of disulfiram induces endometrioid epithelial ovarian cancer cell death via the both apoptosis and cuproptosis pathways

Various therapeutic strategies have been developed to overcome ovarian cancer.However,the prognoses resulting from these strategies are still unclear.In the present work,we screened 54 small molecule compounds approved by the FDA to identify novel agents that could inhibit the viability of human epithelial ovarian cancer cells.Among these,we identified disulfiram(DSF),an old alcohol-abuse drug,as a potential inducer of cell death in ovarian cancer.Mechanistically,DSF treatment significantly reduced the expression of the anti-apoptosis marker Bcell lymphoma/leukemia-2(Bcl-2)and increase the expression of the apoptotic molecules Bcl2 associated X(Bax)and cleaved caspase-3 to promote human epithelial ovarian cancer cell apoptosis.Furthermore,DSF is a newly identified effective copper ionophore,thus the combination of DSF and copper was used to reduce ovarian cancer viability than DSF single treatment.Combination treatment with DSF and copper also led to the reduced expression of ferredoxin 1 and loss of Fe-S cluster proteins(biomarkers of cuproptosis).In vivo,DSF and copper gluconate significantly decreased the tumor volume and increased the survival rate in a murine ovarian cancer xenograft model.Thus,the role of DSF revealed its potential for used as a viable therapeutic agent for the ovarian cancer.

DLAT as a Cuproptosis Promoter and a Molecular Target of Elesclomol in Hepatocellular Carcinoma

Objective Cuproptosis is a novel cell death pathway that was newly discovered in early 2022.However,cuproptosis is still in its infancy in many respects and warrants further research in hepatocellular carcinoma(HCC).This study aimed to analyze the mechanism of cuprptosis in HCC.Methods Herein,the tumor microenvironment infiltration landscape of molecular subtypes was illustrated using GSVA,ssGSEA,TIMER,CIBERSORT,and ESTIMATE algorithms based on the expression profile of cuproptosis-related genes(CRGs)from TCGA and GEO databases.Then,the least absolute shrinkage and selection operator regression method was applied to construct a cuproptosis signature to quantify the cuproptosis profile of HCC.Further,we explored the expression of three hub CRGs in cell lines and clinical patient tissues of HCC by Western blotting,qRT-PCR and immunohistochemistry.Finally,we examined the function of dihydrolipoamide S-acetyltransferase(DLAT)in cuproptosis in HCC by loss-of-function strategy,Western blotting and CCK8 assay.Results Three distinct molecular subtypes were identified.Cluster 2 had the greatest infiltration of immune cells with best prognosis.The cuproptosis signature was indicative of tumor subtype,immunity,and prognosis for HCC,and specifically,a low cuproptosis score foreshadowed good prognosis.DLAT was highly expressed in liver cancer cell lines and HCC tissues and positively correlated with clinical stage and grade.We also found that potent copper ionophore elesclomol could induce cuproptosis in a copper-dependent manner.Selective Cu^(++)chelator ammonium tetrathiomolybdate and downregulating DLAT expression by siRNA could effectively inhibit cuproptosis.Conclusion Cuproptosis and DLAT as a promising biomarker could help to determine the prognosis of HCC and may offer novel insights for effective treatment.

Clinical significance and potential application of cuproptosis-related genes in gastric cancer

BACKGROUND Worldwide,gastric cancer(GC)is a common lethal solid malignancy with a poor prognosis.Cuproptosis is a novel type of cell death mediated by protein lipoylation and may be related to GC prognosis.AIM To offer new insights to predict GC prognosis and provide multiple therapeutic targets related to cuproptosis-related genes(CRGs)for future therapy.METHODS We collected data from several public data portals,systematically estimated the expression level and prognostic values of CRGs in GC samples,and investigated related mechanisms using public databases and bioinformatics.RESULTS Our results revealed that FDX1,LIAS,and MTF1 were differentially expressed in GC samples and exhibited important prognostic significance in The Cancer Genome Atlas(TCGA)cohort.We constructed a nomogram model for overall survival and disease-specific survival prediction and validated it via calibration plots.Mecha-nistically,immune cell infiltration and DNA methylation prominently affected the survival time of GC patients.Moreover,protein-protein interaction network,KEGG pathway and gene ontology enrichment analyses demonstrated that FDX1,LIAS,MTF1 and related proteins play key roles in the tricarboxylic acid cycle and cuproptosis.Gene Expression Omnibus database validation showed that the expression levels of FDX1,LIAS,and MTF1 were consistent with those in the TCGA cohort.Top 10 perturbagens has been filtered CONCLUSION In conclusion,FDX1,LIAS,and MTF1 could serve as potential prognostic biomarkers for GC patients and provide novel targets for immunotarget therapy.

Machine learning algorithm to construct cuproptosis-and immunerelated prognosis prediction model for colon cancer

BACKGROUND Over the past few years,research into the pathogenesis of colon cancer has progressed rapidly,and cuproptosis is an emerging mode of cellular apoptosis.Exploring the relationship between colon cancer and cuproptosis benefits in identifying novel biomarkers and even improving the outcome of the disease.AIM To look at the prognostic relationship between colon cancer and the genes associated with cuproptosis and the immune system in patients.The main purpose was to assess whether reasonable induction of these biomarkers reduces mortality among patients with colon cancers.METHOD Data obtained from The Cancer Genome Atlas and Gene Expression Omnibus and the Genotype-Tissue Expression were used in differential analysis to explore differential expression genes associated with cuproptosis and immune activation.The least absolute shrinkage and selection operator and Cox regression algorithm was applied to build a cuproptosis-and immune-related combination model,and the model was utilized for principal component analysis and survival analysis to observe the survival and prognosis of the patients.A series of statistically meaningful transcriptional analysis results demonstrated an intrinsic relationship between cuproptosis and the micro-environment of colon cancer.RESULTS Once prognostic characteristics were obtained,the CDKN2A and DLAT genes related to cuproptosis were strongly linked to colon cancer:The first was a risk factor,whereas the second was a protective factor.The finding of the validation analysis showed that the comprehensive model associated with cuproptosis and immunity was statistically significant.Within the component expressions,the expressions of HSPA1A,CDKN2A,and UCN3 differed markedly.Transcription analysis primarily reflects the differential activation of related immune cells and pathways.Furthermore,genes linked to immune checkpoint inhibitors were expressed differently between the subgroups,which may reveal the mechanism of worse prognosis and the different sensitivities of chemotherapy.CONCLUSION The prognosis of the high-risk group evaluated in the combined model was poorer,and cuproptosis was highly correlated with the prognosis of colon cancer.It is possible that we may be able to improve patients’prognosis by regulating the gene expression to intervene the risk score.

Assessment of gastric cancer prognosis,immune infiltration based on cuproptosis-related LncRNAs and prediction of traditional Chinese medicine

Objective:Constructing a prognostic model for gastric cancer(GC)based on cuproptosisrelated LncRNAs(CRLs)and predict the traditional Chinese medicine that regulate cuproptosis-related genes(CRGs).Methods:Clinical data and RNA-seq of 443 GC cases were obtained from The Cancer Genome Atlas(TCGA)database,and CRLs were screened by Pearson analysis,Cox regression,and least absolute shrinkage and selection operator(LASSO)regression to construct a risk model to predict GC prognosis,and the nomogram was constructed by combining risk scores and clinical characteristics.The accuracy of the model was validated by the receiver operating characteristic curve,Kaplan-Meier curves and C-index.To assess the correlation of risk scores with immune infiltration,immune checkpoint gene expression and chemotherapy/targeted agents.The Coremine Medical database was applied to predict potential traditional Chinese medicine that regulate CRGs.Results:Risk models for GC were constructed based on the risk scores of seven CRLs(AP001107.9,VCAN-AS1,AC016394.2,LINC02675,AC100814.1,HAGLR,and LINC01094).The AUC of the risk model predicting 1-,3-,and 5-year survival in GC patients was 0.720,0.682,and 0.711,and its prognostic value was better than age,Grade classification,and TNM stage.The AUC of the risk model combining age and TNM stage to predict 1-year survival in GC patients was 0.793.The risk score correlated with the degree of enrichment of immune cells such as tumorinfiltrating lymphocytes and regulatory T cells and the expression of 22 immune checkpoint genes such as LAG3,ICOS,CD28,NRP1 and the sensitivity of 13 chemotherapeutic/targeted agents.There are 58 traditional Chinese medicine with potential regulatory effects on CRGs,mainly for clearing heat and detoxing,promoting blood circulation and relieving pain,which are mainly attributed to the liver,spleen and lung meridians.Spirulina and osthole have potential regulatory effects on FDX1,a key gene in the death mechanism of cuproptosis.Conclusions:A risk signature constructed based on seven CRLs could assess the prognosis and immunity of GC,and Spirulina and Serpentine may have important regulatory efficacy on the mechanism of copper cuproptosis.

Construction and validation of prognostic model of Cuproptosis-related LncRNA in osteosarcoma

Cuproptosis is a newly discovered form of apoptotic process that is thought to play an important role in cancer therapy.Long non-coding RNA(lncRNA)is involved in regulating many physiological and pathological activities of cells.The aim of this study was to investigate the prognostic significance of Cuproptosis-associated lncRNAs in osteosarcoma.Methods:The Gene expression profiling of osteosarcoma samples versus normal samples and corresponding clinical data were downloaded from the public databases UCSC Xena and GTEx,and the cuproptosis gene was obtained from the published literature,the prognostic model of osteosarcoma cuproptosis-related lncRNA was constructed by using coexpression network,minimum absolute contraction and selection algorithm(LASSO)and Cox regression model.Receiver operating characteristic(ROC)curves and nomograms were used to assess the predictive power of the model.Single-sample gene set enrichment analysis(ssGSEA)was used to explore the relationship between osteosarcoma immune cells and function in different risk groups.Results:181 cuproptosis-related lncRNAs were obtained by co-expression analysis of 19 cuproptosis genes collected.Ten lncRNAs were screened out by differential analysis and single-factor Cox analysis.Three cuproptosis-related lncrnas(AC124798.1,AC090152.1,AC090559.1)were screened by Lasso and multivariate Cox regression to construct the prognostic model.Patients were divided into high and low risk groups based on the median risk score.The results of overall survival,risk score distribution and survival status in the lowrisk group were better than those in the high-risk group,and were verified in the internal data.Univariate and multivariate Cox regression analyses showed that risk score was an independent prognostic factor.Nomograms and ROC curves showed that the prognostic model had good predictive ability.The results of ssGSEA suggest that immune cells and function may be inhibited in the high-risk group.Conclusion:The 3 cuproptosis-related lncRNAs may be helpful to guide the prognosis of osteosarcoma patients and provide some theoretical basis for clinical decision.

Cuproptosis-related long non-coding RNAs model that effectively predicts prognosis in hepatocellular carcinoma

BACKGROUND Cuproptosis has recently been considered a novel form of programmed cell death.To date,long-chain non-coding RNAs(lncRNAs)crucial to the regulation of this process remain unelucidated.AIM To identify lncRNAs linked to cuproptosis in order to estimate patients'prognoses for hepatocellular carcinoma(HCC).METHODS Using RNA sequence data from The Cancer Genome Atlas Live Hepatocellular Carcinoma(TCGA-LIHC),a co-expression network of cuproptosis-related genes and lncRNAs was constructed.For HCC prognosis,we developed a cuproptosisrelated lncRNA signature(CupRLSig)using univariate Cox,lasso,and multivariate Cox regression analyses.Kaplan-Meier analysis was used to compare overall survival among high-and low-risk groups stratified by median CupRLSig risk score.Furthermore,comparisons of functional annotation,immune infiltration,somatic mutation,tumor mutation burden(TMB),and pharmacologic options were made between high-and low-risk groups.RESULTS Three hundred and forty-three patients with complete follow-up data were recruited in the analysis.Pearson correlation analysis identified 157 cuproptosis-related lncRNAs related to 14 cuproptosis genes.Next,we divided the TCGA-LIHC sample into a training set and a validation set.In univariate Cox regression analysis,27 LncRNAs with prognostic value were identified in the training set.After lasso regression,the multivariate Cox regression model determined the identified risk equation as follows:Risk score=(0.2659×PICSAR expression)+(0.4374×FOXD2-AS1 expression)+(-0.3467×AP001065.1 expression).The CupRLSig high-risk group was associated with poor overall survival(hazard ratio=1.162,95%CI=1.063-1.270;P<0.001)after the patients were divided into two groups depending upon their median risk score.Model accuracy was further supported by receiver operating characteristic and principal component analysis as well as the validation set.The area under the curve of 0.741 was found to be a better predictor of HCC prognosis as compared to other clinicopathological variables.Mutation analysis revealed that high-risk combinations with high TMB carried worse prognoses(median survival of 30 mo vs 102 mo of low-risk combinations with low TMB group).The low-risk group had more activated natural killer cells(NK cells,P=0.032 by Wilcoxon rank sum test)and fewer regulatory T cells(Tregs,P=0.021)infiltration than the high-risk group.This finding could explain why the low-risk group has a better prognosis.Interestingly,when checkpoint gene expression(CD276,CTLA-4,and PDCD-1)and tumor immune dysfunction and rejection(TIDE)scores are considered,highrisk patients may respond better to immunotherapy.Finally,most drugs commonly used in preclinical and clinical systemic therapy for HCC,such as 5-fluorouracil,gemcitabine,paclitaxel,imatinib,sunitinib,rapamycin,and XL-184(cabozantinib),were found to be more efficacious in the low-risk group;erlotinib,an exception,was more efficacious in the high-risk group.CONCLUSION The lncRNA signature,CupRLSig,constructed in this study is valuable in prognostic estimation of HCC.Importantly,CupRLSig also predicts the level of immune infiltration and potential efficacy of tumor immunotherapy.

Comprehensive analysis of the relationship between cuproptosisrelated genes and esophageal cancer prognosis

BACKGROUND Esophageal cancer is one of the most common malignant tumors of the digestive system,with a 5-year survival rate of 15%to 50%.Cuproptosis,a unique kind of cell death driven by protein lipoylation,is strongly connected to mitochondrial metabolism.The clinical implications of cuproptosis-related genes in esophageal cancer,however,are mainly unknown.AIM To identify cuprotosis-related genes that are differentially expressed in esophageal cancer and investigate their prognostic significance.METHODS With|log fold change|>1 and false discovery rate<0.05 as criteria,the Wilcox test was used to evaluate the differentially expressed genes between 151 tumor tissues and 151 normal esophageal tissues.Cuproptosis-related genes were selected to be linked with prognosis using univariate Cox regression analysis.Genes were separated into high-and low-expression groups based on their cutoff value of gene expression,and the correlation between the two groups and overall survival or progression-free survival was investigated using the log-rank test.The C-index,calibration curve,and receiver operator characteristic(ROC)curve were used to assess a nomogram containing clinicopathological characteristics and cuproptosis-related genes.RESULTS Pyruvate dehydrogenase A1(PDHA1)was found to be highly correlated with prognosis in univariate Cox regression analysis(hazard ratio=22.96,95%confidence interval=3.09-170.73;P=0.002).According to Kaplan-Meier survival curves,low expression of PDHA1 was associated with a better prognosis(log-rank P=0.0007).There was no significant correlation between PDHA1 expression and 22 different types of immune cells.Tumor necrosis factor superfamily member 15(TNFSF15)(P=3.2×10^(-6);r=0.37),TNFRSF14(P=8.1×10^(-8);r=0.42),H long terminal repeat-associating 2(P=6.0×10^(-8);r=0.42)and galectin 9(P=3.1×10^(-6);r=0.37)were all found to be considerably greater in the high PDHA1 expression group,according to an analysis of genes related to 47 immunological checkpoints.The low PDHA1 expression group had significantly lower levels of cluster of differentiation 44(CD44)(P=0.00028;R=-0.29),TNFRSF18(P=1.2×10^(-5);R=-0.35),programmed cell death 1 ligand 2(P=0.0032;R=-0.24),CD86(P=0.018;R=-0.19),and CD40(P=0.0047;R=-0.23),and the differences were statistically significant.We constructed a prognostic nomogram incorporating pathological type,tumor-node-metastasis stage,and PDHA1 expression,and the C-index,calibration curve,and ROC curve revealed that the nomogram’s predictive performance was good.CONCLUSION Cuproptosis-related genes can be used as a prognostic predictor for esophageal cancer patients,providing novel insights into cancer treatment.

The cuproptosis-related gene signature predicts clinical diagnosis,prognosis,and immune microenvironment for glioblastoma multiforme

Background:Glioblastoma multiforme(GBM)is a kind of extremely malignant brain tumor with a poor prognosis.The problem of high recurrence rates and drug resistance has been difficult to solve.Cuproptosis is a newly discovered mode of cell death,which is related to the occurrence and progression of GBM.Methods:We screened TCGA and GEO databases to comprehensively analyze the regulation patterns of cuproptosis in GBM based on cuproptosis-related genes(CRGs).The expression levels,copy number variants(CNV),and mutation of CRGs in GBM were probed.Different regulation patterns of cuproptosis were identified and performed GSVA analysis.Then we constructed a prognostic model based on genes that were differentially expressed in all regulation patterns.The immune cells and tumor microenvironment in different risk groups were compared.Finally,the potential therapeutic drugs were predicted.Results:We identified two key differentially expressed genes under different regulation patterns of cuproptosis and constructed a risk prognostic model.There were significant differences in immune infiltration and drug sensitivity between high and low risk groups.Conclusions:We established a risk prognostic model based on two genes and explored its relationship with immune characteristics,which might help to assess the prognosis and treatment sensitivity to immune and targeted therapy of GBM patients.

Inhalable nanoparticles with enhanced cuproptosis and cGAS-STING activation for synergistic lung metastasis immunotherapy

Due to the insufficient Cu^(+)accumulation,Cu^(+)efflux mechanism,and highly immunosuppressive tumor microenvironment(TME)in lung metastasis,the cuproptosis efficacy is limited.Herein,an inhalable nanodevice(CLDCu)is constructed to successfully overcome the drawbacks of cuproptosis.CLDCu consists of a Cu^(2+)-chitosan shell and low molecular weight heparin-tocopherol succinate(LMWH-TOS,LT)core with disulfiram(DSF)loading.The prepared CLDCu can be inhaled and accumulate in large amounts in lung lesions(63.6%)with 56.5 times higher than intravenous injection.Within tumor cells,the mild acidity triggers the co-release of DSF and Cu2+,thus generating bis(diethyldithiocarbamate)-copper(CuET)to block Cu^(+)efflux protein ATP7B and forming toxic Cu^(+),leading to enhanced cuproptosis.Meanwhile,the released chitosan cooperates with CLDCu-induced cuproptosis to activate stimulator of interferon genes(STING)pathway,which significantly potentiates dendritic cells(DCs)maturation,as wells as evokes innate and adaptive immunity.In lung metastatic mice model,CLDCu is found to induce cuproptosis and reverse the immunosuppressive TME by inhalation administration.Moreover,CLDCu combined with anti-programmed cell death protein ligand-1 antibody(aPD-L1)provokes stronger antitumor immunity.Therefore,nanomedicine that combines cuproptosis with STING activation is a novel strategy for tumor immunotherapy.

CRISPR-Cas9 gene editing strengthens cuproptosis/chemodynamic/ferroptosis synergistic cancer therapy

Copper-based nanomaterials demonstrate promising potential in cancer therapy.Cut efficiently triggers a Fenton-like reaction and further consumes the high level of glutathione,initiating chemical dynamic therapy(CDT)and ferroptosis.Cuproptosis,a newly identified cell death modality that represents a great prospect in cancer therapy,is activated.However,active homeostatic systems rigorously keep copper levels within cells exceptionally low,which hinders the application of cooper nanomaterials-based therapy.Herein,a novel strategy of CRISPR-Cas9 RNP nanocarrier to deliver cuprous ions and suppress the expression of copper transporter protein ATP7A for maintaining a high level of copper in cytoplasmic fluid is developed.The Cu2O and organosilica shell would degrade under the high level of glutathione and weak acidic environment,further releasing RNP and Cut.The liberated Cut triggered a Fenton-like reaction for CDT and partially transformed to Cu2t,consuming intracellular GSH and initiating cuproptosis and ferroptosis efficiently.Meanwhile,the release of RNP effectively reduced the expression of copper transporter ATP7A,subsequently increasing the accumulation of cooper and enhancing the efficacy of CDT,cuproptosis,and ferroptosis.Such tumor microenvironment responsive multimodal nanoplatform opens an ingenious avenue for colorectal cancer therapy based on gene editing enhanced synergistic cuproptosis/CDT/ferroptosis.

Inhalable metal–organic framework-mediated cuproptosis combined with PD-L1 checkpoint blockade for lung metastasis synergistic immunotherapy

Cuproptosis shows enormous application prospects in lung metastasis treatment.However,the glycolysis,Cu^(+)efflux mechanisms,and insufficient lung drug accumulation severely restrict cuproptosis efficacy.Herein,an inhalable poly(2-(N-oxide-N,N-diethylamino)ethyl methacrylate)(OPDEA)-coated copper-based metal–organic framework encapsulating pyruvate dehydrogenase kinase 1 siRNA(siPDK)is constructed for mediating cuproptosis and subsequently promoting lung metastasis immunotherapy,namely OMP.After inhalation,OMP shows highly efficient lung accumulation and long-term retention,ascribing to the OPDEA-mediated pulmonary mucosa penetration.Within tumor cells,OMP is degraded to release Cu2+under acidic condition,which will be reduced to toxic Cu^(+)to induce cuproptosis under glutathione(GSH)regulation.Meanwhile,siPDK released from OMP inhibits intracellular glycolysis and adenosine-5ʹ-triphosphate(ATP)production,then blocking the Cu^(+)efflux protein ATP7B,thereby rendering tumor cells more sensitive to OMP-mediated cuproptosis.Moreover,OMP-mediated cuproptosis triggers immunogenic cell death(ICD)to promote dendritic cells(DCs)maturation and CD8^(+)T cells infiltration.Notably,OMP-induced cuproptosis up-regulates membrane-associated programmed cell death-ligand 1(PD-L1)expression and induces soluble PD-L1 secretion,and thus synergizes with anti-PD-L1 antibodies(aPD-L1)to reprogram immunosuppressive tumor microenvironment,finally yielding improved immunotherapy efficacy.Overall,OMP may serve as an efficient inhalable nanoplatform and afford preferable efficacy against lung metastasis through inducing cuproptosis and combining with aPD-L1.

Cuproptosis mediates copper-induced testicular spermatogenic cell death

Cuproptosis,a novel mechanism of programmed cell death,has not been fully explored in the context of spermatogenic cells.Thisstudy investigated the potential involvement of cuproptosis in spermatogenic cell death using a mouse model of copper overload.Sixty male Institute of Cancer Research(ICR)mice were randomly divided into four groups that received daily oral gavage withsodium chloride(control)or copper sulfate(CuSO_(4))at 50 mg kg^(−1),100 mg kg^(−1),or 200 mg kg^(−1),for 42 consecutive days.Micesubjected to copper overload exhibited a disruption in copper homeostasis.Additionally,significant upregulated expression of keycuproptosis factors was accompanied by a significant rise in the rates of testicular tissue cell apoptosis.Immunohistochemicalanalysis revealed the presence of ferredoxin 1(Fdx1)in Sertoli cells,Leydig cells,and spermatogenic cells at various stages oftesticular development,and the Fdx1-positive staining area was significantly increased in copper-overloaded mice.Mitochondrialdysfunction and decreased adenosine triphosphate levels were also observed,further implicating mitochondrial damage undercuproptosis.Further analyses revealed pathological lesions and blood−testis barrier destruction in the testicular tissue,accompaniedby decreased sperm concentration and motility,in copper-overloaded mice.In summary,our results indicate that copper-overloadedmice exhibit copper homeostasis disorder in the testicular tissue and that cuproptosis participates in spermatogenic cell death.These findings provide novel insights into the pathogenic mechanisms underlying spermatogenic cell death and provide initialexperimental evidence for the occurrence of cuproptosis in the testis.