A novel inflammation-associated prognostic signature for clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) are typically situated in a complex inflammatory and immune microenvironment, which has been reported to contribute to the unfavorable prognosis of patients with ccRCC. There would be beneficial clinical implications for elucidating the roles of its molecular characteristics in the inflammatory microenvironment. This is because it would facilitate the development of reliable biomarkers for pre-stratification prior to the designation of individualized treatment strategies. In the present study, RNA-sequencing data from 607 patients were retrospectively analyzed to elucidate the profile of inflammatory molecules. Based on this, an inflammatory prognostic signature (IPS) was developed and further validated using clinical ccRCC samples. Subsequently, the associated mechanisms in terms of the immune microenvironment and molecular pathways were then investigated. This proposed IPS was found to exhibit superior accuracy compared with the criterion of a good prognostic model for the prediction of patient prognosis from ccRCC [area under the receiver operating characteristic curve (AUC)=0.811] in addition to being an independent factor for prognostic risk stratification [hazard ratio: 11.73 (95% CI, 26.98-5.10); log-rank test, P<0.001]. Pathologically, ccRCC cells identified as high-risk according to their IPS presented with a more malignant tumor structure, including voluminous eosinophilic cytoplasm, acinar/lamellar/tubular growth patterns and atypic nuclei. High-risk ccRCC also exhibited higher infiltration levels by four types of immune cells, including T regulatory cells, but lower infiltration levels by mast cells. Pathways associated with immune-inflammation interaction, including the IL-17 pathway, were found to be upregulated in IPS-identified high-risk ccRCC. Furthermore, by combining the IPS with clinical factors, an integrated prognostic index was developed and validated for increasing the accuracy of patient risk-stratification for ccRCC (AUC=0.911). In conclusion, the complex regulatory mechanisms and molecular characteristics involved in ccRCC-inflammation interaction, coupled with their prognostic potential, were systematically elucidated in the present study. This may have important implications in furthering the understanding into the molecular mechanisms underlying this ccRCC-inflammation interaction, which can in turn be exploited for identifying high-risk patients with ccRCC prior to designing their clinical treatment strategy.     

Construction and validation of a m6A RNA methylation and ferroptosis-related prognostic model for pancreatic cancer by integrated bioinformatics analysis

BackgroundBoth N6-methyladenosine (m6A) ribonucleic acid (RNA) methylation and ferroptosis regulators are demonstrated to have significant effects on the malignant clinicopathological characteristics of pancreatic adenocarcinoma (PAAD) patients. However, the currently available clinical indexes are not sufficient to predict precise prognostic outcomes pf PAAD patients accurately. This study aims to examine the clinicopathologic features of m6A RNA methylation and ferroptosis regulators in predicting the outcomes of different types of cancer.MethodsAs the foundation for this research, the differentially expressed genes (DEGs) between PAAD tissues and adjacent normal tissues were first identified. Next, dimensional reduction analysis (DCA) based on m6A RNA methylation regulators and ferroptosis regulators were performed and DEGs between good/poor prognosis PAAD patient clusters were identified. DEGs were then screened by Cox analysis, and finally a risk signature was established by least absolute shrinkage and selection operator (LASSO) analyses. The prediction model based on risk score was further evaluated by a validation set from Gene Expression Omnibus (GEO) database.ResultsIn total, 4 m6A RNA methylation regulator genes and 29 ferroptosis regulator genes were found to have close causal relationships with the prognosis of PAAD, and a risk score with 3 m6A methylation regulators (i.e., IGF2BP2, IGF2BP3, and METTL16) and 4 ferroptosis regulators (i.e., ENPP2, ATP6V1G2, ITGB4, and PROM2) was constructed and showed to be highly involved in PAAD progression and could serve as effective markers for prognosis with AUC value equaled 0.753 in training set and 0.803 in validation set.ConclusionsThe combined prediction model, composed of seven regulators of m6A methylation and ferroptosis, in this study more effectively reflects the progression and prognosis of PAAD than previous single genome or epigenetic analysis. Our study provides a broader perspective for the subsequent establishment of prognostic models and the patients may benefit from more precision management.     

肺腺癌m6A甲基化调控因子相关预后风险模型的构建及临床意义

目的 构建基于N6-甲基腺苷(N6-methyladenosine,m6A)甲基化调控因子的肺腺癌预后风险模型,为肺腺癌的预后评估提供科学依据。方法 从TCGA数据库下载mRNA表达和临床数据。使用R软件对24种m6A甲基化调控因子进行差异表达分析。利用单因素Cox回归分析初步筛选出与生存相关的m6A调控因子,在此基础上通过Lasso回归进一步筛选纳入模型的变量,将Lasso回归得到的m6A调控因子用于构建Cox回归模型并计算每个样本的风险评分。qRT-PCR检测模型基因HNRNPC及IGF2BP1在正常肺上皮细胞(REAS-2B)及肺腺癌细胞株(A549、H1299、PC9)中的表达。采用TCGA数据库中配对组织的差异分析检测模型基因在肺腺癌及正常肺组织的差异表达。使用Kaplan-Meier生存曲线及ROC曲线对模型效能进行评估。通过热图分析不同风险组的临床特征,并结合其他临床参数进行单因素和多因素Cox回归分析对风险模型的独立预后性进行检验。结果 19个m6A甲基化调控因子在肺腺癌和正常组织中显著差异表达。单因素Cox回归分析发现4个肺腺癌生存显著相关的m6A调控因子(P＜0.01),进一步采用Lasso回归联合Cox回归算法构建了包含2个基因(IGF2BP1、HNRNPC)的预后风险模型。qRT-PCR结果显示HNRNPC和IGF2BP1相对表达量在肺腺癌细胞株A549、H1299和PC-9中高于正常肺上皮细胞REAS-2B,差异具有统计学意义(P＜0.01)。TCGA数据库的59对肺腺癌及相邻的正常肺组织的匹配差异分析发现HNRNPC和IGF2BP1在肺腺癌中表达高于正常肺组织,差异具有统计学意义(P＜0.001)。生存曲线分析显示相比低风险组患者,高风险患者总体生存期明显降低(P＜0.01)。ROC曲线结果表明模型可以较好地预测肺腺癌患者预后(AUC=0.724)。多因素Cox回归分析显示该预后风险模型可作为独立预后因素(HR=2.357,P＜0.001)对肺腺癌患者的预后进行预测。结论 本研究构建了基于m6A甲基化调控因子的预后风险评估模型,且该模型具有较好的预测能力,对制定合理及有效的个体化治疗方案具有潜在的参考价值。     

Hepcidin as a prognostic biomarker in clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is a common malignancy of urologic neoplasms. Hepcidin is a pivotal modulator of iron metabolism involved in human cancers; however, the biological significance of hepcidin in ccRCC remains to be fully understood. Therefore, in this study, we evaluated the expression profiles of hepcidin in ccRCC from several public databases and found that hepcidin expression was upregulated in ccRCC, which was further validated in ccRCC cell lines, clinical samples, and tissue microarray (TMA) quantitative real-time PCR and immunohistochemistry. In addition, we found that the expression level of hepcidin was correlated with the age, T stage and pathologic stage of patients. Furthermore, hepcidin promoter methylation was significantly associated with the worse poor clinical parameters of ccRCC patients, and hepcidin was an independent prognostic factor. Mechanistically, enrichment analysis revealed that hepcidin participated in the immune-related and metabolism-related pathways. Hepcidin was positively correlated with not only immune infiltration and immune checkpoints but also tumor mutation burden and cytotoxic T lymphocyte. Finally, we validated the positive correlation of hepcidin with the marker of macrophage (CD68) in the TMA. Our findings provide insights into understanding the function and its underlying mechanism of hepcidin in ccRCC and suggest that hepcidin might serve as a potential predictive biomarker of response to immunotherapy and the prognosis of patients with ccRCC.     

Upregulation of COL6A1 is predictive of poor prognosis in clear cell renal cell carcinoma patients

Background: The extracellular matrix (ECM) is reported to play an important role in tumorigenesis and progression. Collagen VI is an important ECM protein. In this study, we investigated the potential role of the COL6A1 gene, which encodes the α1 polypeptide of collagen VI, in the biological functions involved in the progression and outcome of clear cell renal cell carcinoma (ccRCC).Materials and methods: A total of 288 ccRCC patients who underwent radical nephrectomy (RN) or nephron sparing nephrectomy (NSS) at Fudan University Shanghai Cancer Center (FUSCC) were enrolled. Total RNA was extracted from frozen samples obtained from the tissue bank of FUSCC and expression of COL6A1 was determined by qRT-PCR. The clinical relationship between COL6A1 expression and ccRCC prognosis was analyzed. These data were then validated in the Cancer Genome Atlas (TCGA) cohort. We also investigated the effect of COL6A1 overexpression in a xenografted tumor model in nude mice in vivo.Results: In multivariate analysis of TCGA cohorts, COL6A1 high expression was predictive of poor prognosis in ccRCC patients’ overall survival (OS) (HR: 2.588 95%CI 1.616–4.146) and disease free survival(DFS) (HR: 3.106 95%CI 1.534–6.288). In FUSCC cohorts, after adjusted for relevant factors, the COL6A1 expression indicates poor prognosis in ccRCC patients’s OS (HR 2.211; 95% CI, 1.360–8.060) and DFS (HR 3.052; 95%CI, 1.500–6.210). COL6A1 overexpression promoted tumor growth in xenografted nude mice.Conclusion: Increased COL6A1 expression correlates with poor prognosis in ccRCC patients. Moreover, COL6A1 stimulates tumor growth in vivo.     

基于数据挖掘分析PEG3基因在肾透明细胞癌中的表达及预后意义

目的:阐述印迹基因PEG3在肾细胞癌中的表达及意义。方法:利用Oncomine数据库及GEPIA分析PEG3在肾透明细胞癌组织中的表达情况;利用KM-Plotter做PEG3与肾透明细胞癌患者生存期的相关性分析,经MethHC分析PEG3启动子区甲基化水平,String-DB数据库分析PEG3相互作用蛋白;通过The Human Protein Atlas分析PEG3蛋白在肾透明细胞癌中的蛋白表达情况。结果:在mRNA水平上,PEG3在肾透明细胞癌中较正常肾组织显著低表达,表达水平与肾癌预后呈负相关（Log-rank P=0.001 1）,PEG3蛋白在肾透明细胞癌中也较正常肾脏组织低表达,且其启动子甲基化水平则显著增高。与PEG3相关蛋白有NLRP7、MEST、SIAH1、PEG10、CDC25B等,可能参与细胞有丝分裂周期调节及遗传印迹等细胞功能。结论:通过肿瘤基因数据库信息挖掘发现, PEG3在肾透明细胞癌组织中呈低表达,且与患者生存期有关,将为后续的肿瘤研究提供重要的理论依据。     

肾透明细胞癌组织Fibulin-3表达及其临床意义

目的 探讨Fibulin-3在肾透明细胞癌(clear cell renal cell carcinoma,CCRCC)中的表达及其临床意义.方法 收集中山大学附属第一医院2000-04-01-2007-04-01手术切除并经病理确诊的157例CCRCC患者的石蜡标本及相关临床病理资料.其中男10例,女56例,平均年龄51.7岁.制作组织芯片,应用免疫组织化学方法检测Fibulin-3蛋白的表达情况,根据评分结果将RCC患者分为Fibulin-3高表达和低表达两组,并分析其与患者多种临床参数的相关性.结果 Fibulin-3在157例CCRCC组织中的阳性表达率为100.0％,高表达58例,低表达99例.Fibulin-3蛋白表达与CCRCC患者的性别、年龄、肿瘤大小、Fuhrman分级和组织学类型等临床病理参数无相关性,P＞0.05.Kaplan-Meier生存分析显示,Fibulin-3高表达与肾癌患者的预后相关,P=0.003;多因素Cox回归分析表明,Fibulin-3是CCRCC患者生存时间的独立影响因素,HR=2.30,95％CI为1.278～4.140,P=0.005.结论 Fibulin-3高表达与CCRCC患者的不良预后相关,提示Fibulin-3是预测CCRCC患者预后的潜在分子标志.     

Construction and validation of an N6-methyladenosine-associated prognostic signature in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is among the most common types of cancer that threat the public health worldwide. N6-methyladenosine (m6A) RNA methylation is associated with cancer initiation and progression, and is dynamically regulated by m6A RNA methylation-associated genes. However, little is known about the expression status and the prognostic value of m6A-associated genes in HCC. The present study aimed to identify the expression profiling pattern and clinical significance of m6A-associated genes in HCC. Consensus clustering analysis was performed to identify the clusters of HCC with different clinical outcomes. A prognostic signature built by the least absolute shrinkage and selection operator Cox regression model was utilized to discover subtypes associated with different clinical outcomes of patients with HCC in the discovery cohort from The Cancer Genome Atlas. The differences between subgroups were characterized in terms of epigenetic dysregulation and somatic mutation frequencies. The International Cancer Genome Consortium cohort and two independent cohorts from the meta-Gene Expression Omnibus database were used for external validation. Most of the m6A-associated genes were upregulated and involved in the prognosis and malignancy of HCC. A four-gene prognostic signature revealed two HCC subtypes (namely, high- and low-risk group) that was associated with different clinical outcomes. Patients in the high-risk group were accompanied with increased epigenetic silencing and significant mutations in TP53 and FLG, while ALB was frequently mutated in the low-risk group. In conclusion, an m6A-based signature was constructed to predict the prognosis of patients with HCC, which may provide a tool for reliable prognosis assessment for clinicians, and aid clinical treatment decision-making.     

血清IL-22在肾透明细胞癌患者预后分析中的应用价值

目的:研究IL-22在肾透明细胞癌（clear cell renal cell carcinoma,ccRCC）患者血清中的表达情况、临床意义及其在患者预后分析中的应用价值。方法:收集168例ccRCC患者的血清,同时以30例健康志愿者血清作为对照。采用ELISA法检测两组对象血清IL-22的表达水平。生存分析探讨IL-22表达水平与患者预后情况的关系。Cox比例风险模型分析影响患者生存时间的危险因素。结果:ccRCC患者血清中的IL-22水平明显高于健康志愿者。对ccRCC患者随访5~63个月,平均（45±12.4）个月,IL-22高水平组的无进展生存率及总体生存率明显低于低水平组。多因素分析结果表明,血清IL-22表达水平、肿瘤直径、肿瘤分化程度及肿瘤TNM分期是影响ccRCC患者生存期的独立危险因素。结论:ccRCC患者血清IL-22表达水平偏高,且IL-22高水平表达患者预后较差。血清IL-22表达水平在ccRCC患者的预后判断中具有较好的应用价值。     

Silent information regulator 2 promotes clear cell renal cell carcinoma progression through deacetylation and small ubiquitin-related modifier 1 modification of glucose 6-phosphate dehydrogenase

The regulatory relationship between silent information regulator 2 (SIRT2) and glucose 6-phosphate dehydrogenase (G6PD) in clear cell renal cell carcinoma (ccRCC) is still unclear. The present study aimed to explore the function of SIRT2 and its regulatory effect on G6PD in ccRCC. The Cancer Genome Atlas data mining of SIRT2 was first analyzed. Quantitative real-time PCR and western blot analyses were used to assess the mRNA and protein expression levels, respectively. Cell viability, colony formation, cell cycle, cell apoptosis, and TUNEL assays and EdU staining were used to investigate the roles of SIRT2 in ccRCC proliferation and apoptosis. The coimmunoprecipitation (Co-IP) assay was used to analyze the association between SIRT2 and G6PD in ccRCC cells. Quantitative Co-IP assay was used to detect the levels of G6PD ubiquitination and small ubiquitin-related modifier 1 (SUMO1). An in vivo experiment was also carried out to confirm in vitro findings. The results indicated that SIRT2 promoted ccRCC proliferation and inhibited apoptosis by regulating cell cycle and apoptosis related proteins. Silent information regulator 2 interacted with G6PD, facilitated its activity through deacetylation, and increased its stability by reducing its ubiquitination and enhancing its SUMO1 modification. Silent information regulator 2 also promoted ccRCC tumor development in vivo. Taken together, the present study indicated that SIRT2 promoted ccRCC progression by increasing G6PD activity and stability, and it could be a potential new diagnostic and therapeutic target for ccRCC.     

代谢综合征与肾透明细胞癌相关性研究

目的 代谢综合征(metabolic syndrome,MS)是一组临床症候群,MS及其相关组分与癌症发生发展及病理特征具有密切关系.本研究旨在分析MS及其相关组分与肾透明细胞癌(clear cell cenal cell carcinoma,CCRCC)分期、分级及肿瘤大小的相关性.方法 回顾性分析2013-01-01-2015-12-30于山西医科大学第一医院就诊且病理诊断为CCRCC的375例患者的临床资料,包括年龄、性别、身高、体质量、血压、空腹血糖、生化结果、病理分期分级和肿瘤大小等.计数资料采用x2检验,计量资料以(x)±s表示,组间比较采用t检验,多因素分析采用Logistic回归分析.结果 MS组56例患者,其中男性患者32例,女性患者24例;非MS组319例患者,其中男性患者206例,女性患者113例.男女患者比较,差异无统计学意义,P=0.287.MS组与非MS组相比,年龄、吸烟、饮酒等差异无统计学意义,P值分别为0.100、0.691和0.269;而BMI指数、收缩压、空腹血糖、TG、HDL-C等差异均有统计学意义,均P＜0.001.在病理特点方面,MS与非MS相比,CCRCC病理分期(P=0.018)、分级(P=0.026)及肿瘤大小(P=0.026),差异均有统计学意义.MS相关疾病与CCRCC分期分析,糖尿病(P＜0.001)、高血压(P=0.015)、血脂紊乱(P=0.006)与CCRCC的分期有关.结论 CCRCC合并MS者病理分期较高、分级较低、肿瘤更大,糖尿病、高血压和血脂紊乱都可增加CCRCC的病理分期.     

基于TCGA数据库探究肾透明细胞癌关键基因的表达及预后作用

目的:探究肾透明细胞癌中关键基因的表达及预后作用,寻找潜在治疗靶点。方法:从TCGA数据库下载肾透明细胞癌mRNA的表达数据,通过Rstudio软件分析肿瘤与正常组织间差异表达基因,对差异表达基因进行富集分析、蛋白-蛋白相互作用网络构建,并分析出关键基因,最后对关键基因进行预后分析。结果:得到1 855个差异表达基因,其中有1 207个是上调的,648个是下调的,富集分析发现差异基因主要与信号转导、物质代谢、免疫反应等信号通路相关。筛选出10个关键基因中有6个存在预后价值。结论:筛选出的差异基因及信号通路可以帮助我们探索肾透明细胞癌发病的分子机制,同时为靶向治疗的研究提供潜在的理论依据。     

RhoB在肾透明细胞癌中的表达及临床意义

目的:探讨RhoB在肾透明细胞癌中的表达及临床意义.方法:采用实时定量PCR、Western blot和免疫组化方法检测RhoB在肾透明细胞癌组织中的表达情况.选取60例肾透明细胞癌组织标本,详细收集病人临床病理资料,通过免疫组化方法检测RhoB在肾透明细胞癌组织中的蛋白表达水平,并分析RhoB的蛋白水平与临床病理资料的关系.结果:与对应瘤旁肾组织相比,RhoB的mRNA及蛋白表达水平在肾透明细胞癌组织标本中明显降低;RhoB的蛋白表达水平在不同年龄、性别组间差异无统计学意义(P>0.05),而在肿瘤直径大小、T分期、临床分期和组织分级间差异有显著统计学差异(P<0.05).结论:RhoB的表达降低可能在肾透明细胞癌的肿瘤发生中发挥作用,且其表达下降可能与肾透明细胞癌的恶性进展有关.     

Sirtuin 6 regulates the proliferation and survival of clear cell renal cell carcinoma cells via B-cell lymphoma 2

Sirtuin 6 (SIRT6) is a member of the third family of longevity proteins (SIRTs) that is involved in the development of different types of cancer. However, the potential role of SIRT6 in clear cell renal cell carcinoma (ccRCC) and its molecular mechanism have not yet been fully elucidated. Therefore, the present study aimed to investigate the association between SIRT6 and ccRCC, and to further examine the underlying mechanism of its effect on ccRCC proliferation, using bioinformatics analysis, and in vitro and in vivo experiments. The results of the present study demonstrated that SIRT6 was upregulated in ccRCC tissues. In addition, bioinformatics analysis revealed that high SIRT6 expression was closely associated with poor prognosis of patients with ccRCC. In vitro experiments demonstrated that silencing SIRT6 expression in ccRCC-derived 769-P and 786-O cells significantly inhibited their proliferation, migration and invasion. Consistent with these results, in vivo assays demonstrated that SIRT6 knockdown markedly attenuated tumor growth arising from 769-P cells. Furthermore, depletion of SIRT6 enhanced the sensitivity of ccRCC cells to cisplatin. Notably, silencing SIRT6 expression decreased B-cell lymphoma 2 (Bcl-2) expression and increased Bax expression, respectively. Taken together, these results suggest that SIRT6 acts as a proto-oncogene in ccRCC through the augmentation of the Bcl-2-dependent pro-survival pathway, and may be used as a therapeutic target for patients with ccRCC.     

基于生物信息学数据分析ELTD1在肾透明细胞癌组织中的表达、甲基化水平及其临床意义

目的：探讨表皮生长因子蛛毒素受体7穿膜结构域蛋白1(ELTD1)在肾透明细胞癌（ccRCC）组织中的表达、甲基化水平及其与患者临床病理特征和预后的相关性。方法：通过公共数据库分析ccRCC组织中ELTD1表达和甲基化的差异水平,探讨ELTD1表达水平与患者临床病理特征和预后的相关性。通过TIMER2.0数据库评估ccRCC免疫细胞浸润,筛选ELTD1相关免疫检查点基因,进行GO功能和KEGG通路富集分析,通过基因共表达分析、筛选与ELTD1相关的基因。结果：与癌旁组织比较,ELTD1在ccRCC组织中呈高表达（P<0.05）。TCGA-KIRC队列中,ELTD1的甲基化水平与其表达呈负相关（R=-0.37,P<0.01）。ELTD1转录表达在ccRCC患者年龄、T分期、M分期、临床分期及病理分级组间存在显著差异（均P<0.01）,且高表达ELTD1与较长的OS和PFS密切相关（HR=0.55、0.63,均P<0.01）,ELTD1高表达是ccRCC的独立保护因素。ELTD1表达与B细胞、CD4+T细胞、CD8+T细胞、巨噬细胞和嗜中性粒细胞的免疫浸润呈显著负相关...     

Pericyte coverage of differentiated vessels inside tumor vasculature is an independent unfavorable prognostic factor for patients with clear cell renal cell carcinoma

BACKGROUND.

The objective of this study was to evaluate the effect of pericyte coverage (PC) of differentiated tumor microvessels on the prognosis of patients with clear cell renal cell carcinoma (CCRCC).

METHODS.

Samples from 2 cohorts of patients with CCRCC (101 Asian patients and 524 US patients) were prepared using 2 different histologic approaches: routine sectioning versus tissue microarray. Then, the samples were immunohistochemically doubled‐stained for a pericyte marker (alpha smooth muscle actin [α‐SMA]) and a differentiated vessel marker (cluster of differentiation 34 [CD34]), followed by multispectral image capturing and computerized image analyses to quantify the microvessel density (MVD) and the PC of differentiated vessels. The correlations of PC and the MVD:PC ratio with clinicopathologic characteristics were analyzed.

RESULTS.

There was an inverse correlation between differentiated MVD and PC. Higher PC correlated with more aggressive clinicopathologic characteristics of CCRCC in both cohorts, including more advanced T‐classification, higher pathologic grades, and the occurrence of tumor necrosis. The MVD:PC ratio was an independent favorable prognostic factor for overall and recurrence‐free survival in the Asian cohort and for recurrence‐free survival in the US cohort. PC also was an independent prognostic factor, with higher PC predicting a poorer outcome. The combination of PC and MVD was better at distinguishing the outcome of patients with CCRCC. PC combined with differentiated MVD or with the MVD:PC ratio provided additional, independent prognostic information to the Leibovich risk model, and that information was used to generate improved risk models.

CONCLUSIONS.

The authors consistently observed that higher PC was correlated with more aggressive clinicopathologic characteristics. PC was an independent unfavorable prognostic factor. The authors concluded that pericytes should be considered for therapeutic targeting. Cancer 2013. © 2012 American Cancer Society.     

EFHD1, a novel mitochondrial regulator of tumor metastasis in clear cell renal cell carcinoma

The biological function of many mitochondrial proteins in mechanistic detail has not been well investigated in clear cell renal cell carcinoma (ccRCC). A seven-mitochondrial-gene signature was generated by Lasso regression analysis to improve the prediction of prognosis of patients with ccRCC, using The Cancer Genome Atlas and Clinical Proteomic Tumor Analysis Consortium cohort. Among those seven genes, EFHD1 is less studied and its role in the progression of ccRCC remains unknown. The decreased expression of EFHD1 was validated in clinical samples and was correlated with unfavorable outcome. Overexpression of EFHD1 in ccRCC cells resulted in the reduction of mitochondrial Ca²⁺, and the inhibition of cell migration and invasion in vitro and tumor metastasis in vivo. Mechanistically, EFHD1 physically bound to the core mitochondrial calcium transporter (mitochondrial calcium uniporter, MCU) through its N-terminal domain. The interaction between EFHD1 and MCU suppressed the uptake of Ca²⁺ into mitochondria, and deactivated the Hippo/YAP signaling pathway. Further data revealed that the ectopic expression of EFHD1 upregulated STARD13 to enhance the phosphorylation of YAP protein at Ser-127. The knockdown of STARD13 or the overexpression of MCU partly abrogated the EFHD1-mediated induction of phosphorylation of YAP at Ser-127 and suppression of cell migration. Taken together, the newly identified EFHD1–MCU–STARD13 axis participates in the modulation of the Hippo/YAP pathway and serves as a novel regulator in the progression of ccRCC.