four-miRNA Signature Research Articles

MicroRNA expression profiles in muscle-invasive bladder cancer: identification of a four-microRNA signature associated with patient survival.

Bladder cancer ranks the second most common genitourinary tract cancer, and muscle-invasive bladder cancer (MIBC) accounts for approximately 25 % of all bladder cancer cases with high mortality. In the current study, with a total of 202 treatment-naïve primary MIBC patients identified from The Cancer Genome Atlas dataset, we comprehensively analyzed the genome-wide microRNA (miRNA) expression profiles in MIBC, with the aim to investigate the relationship of miRNA expression with the progression and prognosis of MIBC, and generate a miRNA signature of prognostic capabilities. In the progression-related miRNA profiles, a total of 47, 16, 3, and 84 miRNAs were selected for pathologic T, N, M, and histologic grade, respectively. Of the eight most important progression-related miRNAs, four (let-7c, mir-125b-1, mir-193a, and mir-99a) were significantly associated with survival of patients with MIBC. Finally, a four-miRNA signature was generated and proven as a promising prognostic parameter. In summary, this study identified the specific miRNAs associated with the progression and aggressiveness of MIBC and a four-miRNA signature as a promising prognostic parameter of MIBC.

Identification of a plasma four-microRNA panel as potential noninvasive biomarker for osteosarcoma.

BackgroundCirculating microRNAs (miRNAs) are emerging as promising biomarkers for human cancer. Osteosarcoma is the most common human primary malignant bone tumor in children and young adults. The objective of this study was to investigate whether circulating miRNAs in plasma could be a useful biomarker for detecting osteosarcoma and monitoring tumor removal dynamics.MethodsPlasma samples were obtained from 90 patients before surgery, 50 patients after one month of surgery, and 90 healthy individuals. The study was divided into three steps: First, initial screening of the profiles of circulating miRNAs in pooled plasma samples from healthy controls and pre-operative osteosarcoma patients using a TaqMan low density array (TLDA). Second, evaluation of miRNA concentration in individual plasma samples from 90 pre-operative osteosarcoma patients and 90 healthy controls by a quantitative real time PCR (qRT-PCR) assay. Third, evaluation of miRNA concentration in paired plasma samples from 50 pre- and post-operative osteosarcoma patients by qRT-PCR assay.ResultsFour plasma miRNAs including miR-195-5p, miR-199a-3p, miR-320a, and miR-374a-5p were significantly increased in the osteosarcoma patients. Receiver operating characteristics curve analysis of the combined populations demonstrated that the four-miRNA signature could discriminate cases from controls with an area under the curve of 0.9608 (95% CI 0.9307-0.9912). These 4 miRNAs were markedly decreased in the plasma after operation. In addition, circulating miR-195-5p and miR-199a-3p were correlated with metastasis status, while miR-199a-3p and miR-320a were correlated with histological subtype.ConclusionsOur data suggest that altered levels of circulating miRNAs might have great potential to serve as novel, non-invasive biomarkers for osteosarcoma.

A serum microRNA signature predicts tumor relapse and survival in triple-negative breast cancer patients.

Triple-negative breast cancers (TNBC) are associated with high risk of early tumor recurrence and poor outcome. Common prognostic biomarkers give very restricted predictive information of tumor recurrences in TNBC. Human serum contains stably expressed microRNAs (miRNAs), which have been discovered to predict prognosis in patients with cancer. The purpose of this study was to identify circulating biomarkers able to predict clinical outcome in TNBC. We performed genome-wide serum miRNA expression and real-time PCR analyses to investigate the ability of miRNAs in predicting tumor relapse in serum samples from 60 primary TNBC. Patients were divided into training and testing cohorts. By Cox regression analysis, we identified a four-miRNA signature (miR-18b, miR-103, miR-107, and miR-652) that predicted tumor relapse and overall survival. This miRNA signature was further validated in an independent cohort of 70 TNBC. A high-risk signature score was developed and significantly associated with tumor recurrence and reduced survival. Multivariate Cox regression models indicated that the risk score based on the four-miRNA signature was an independent prognostic classifier of patients with TNBC. This signature may serve as a minimally invasive predictor of tumor relapse and overall survival for patients with TNBC. This prediction model may ultimately lead to better treatment options for patients with TNBC.

MiRNA expression profile reveals a prognostic signature for esophageal squamous cell carcinoma

In this study, we analyzed the microRNA (miRNA) expression profile of 119 paired ESCC samples by microarray and identified a four-miRNA signature that predicted patient survival. The signature derived from the training set (n=60) had a good prognostic value in the test set (n=59) and the independent cohort (n=58), indicating the replicability of its prognostic value. Furthermore, the stratified analysis showed that the signature could predict the survival of TNM stage II and stage III patients, indicating that the four-miRNA signature could help to more accurately predict ESCC patient survival.

Identification of four serum microRNAs from a genome-wide serum microRNA expression profile as potential non-invasive biomarkers for endometrioid endometrial cancer

Serum microRNAs (miRNAs), with their remarkable stability and unique concentration profiles in patients with various diseases, are promising non-invasive biomarkers for tumor detection. The present study investigated the altered profiles of serum microRNAs in patients with endometrioid endometrial cancer (EEC) in order to predict the malignancy of the disease at a relatively early stage. TaqMan® low-density arrays (TDLAs) were used to perform an analysis in the initial screening phase using serum samples pooled from seven EEC patients and 20 controls. The differential expression was validated using a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (qRT-PCR) in samples taken from 26 EEC patients and 22 age- and gender-matched healthy controls. The data obtained from the TLDAs demonstrated that 22 serum miRNAs were markedly upregulated in the EEC patients compared with the controls. The qRT-PCR analysis further identified a profile of four serum miRNAs (miR-222, miR-223, miR-186 and miR-204) as a fingerprint for EEC detection. The area under the receiver operating characteristic (ROC) curve of this four-serum miRNA signature was 0.927, which was markedly higher than that of carbohydrate antigen 125 (CA-125; 0.673). The four-miRNA signature identified by genome-wide serum miRNA expression profiling analysis provides a novel, non-invasive approach for EEC diagnosis.

Serum microRNA profiling and breast cancer risk: the use of miR-484/191 as endogenous controls

It has been demonstrated that there are abundant stable microRNAs (miRNAs) in plasma/serum, which can be detected and are potentially disease specific. However, the lack of suitable endogenous controls for serum miRNA detection is the restriction for the widely usage of this kind of biomarkers and for the between-laboratory comparison of the findings. We first systematically screened for endogenous control miRNAs (ECMs) by testing 10 pooling samples (using both Solexa sequencing and TaqMan low density array) and 50 individual samples (using quantitative reverse transcription-PCR) of different cancer traits and healthy controls. Then we assessed serum miRNAs used as potential biomarkers for breast cancer risk prediction based on a two-stage case-control analysis, including 48 breast cancer patients and 48 controls for the discovery stage and 76 breast cancer patients and 76 controls for validation. We identified two candidate ECMs (miRNA-191 and miRNA-484). Normalized by the two ECMs, we found four miRNAs (miR-16, miR-25, miR-222 and miR-324-3p) that were consistently differentially expressed between breast cancer cases and controls. The area under the receiver operating characteristic curve is 0.954 for the four-miRNA signature in the discovery stage (sensitivity = 0.917 and specificity = 0.896) and 0.928 in the validation stage (sensitivity = 0.921 and specificity = 0.934). In conclusion, the four-miRNA signature from serum may serve as a non-invasive prediction biomarker for breast cancer. Furthermore, we proposed the combination of miRNA-484 and miRNA-191 as endogenous control for serum miRNA detection, at least for most common cancers.

Abstract 161: Plasma-based biomarkers for breast cancer diagnosis

Abstract Introduction It has been demonstrated that there are abundant, stable microRNAs (miRNAs) in plasma/serum, which can be detected and is potentially disease-specific. In this study, we aimed to assess whether serum/plasma miRNAs may be used as potential biomarkers for breast cancer diagnosis. Methods We used systematically Solexa sequencing as well as TaqMan Low Density Array followed by individual quantitative reverse transcription polymerase chain reaction assays to screen plasma miRNAs in 48 breast cancer patients and 48 age-matched controls of Chinese origin for the discovery stage and validated in an internal validation set (50 patients and 50 controls in Chinese) and in an external validation set (24 patients and 24 controls in Caucasians). Results Four miRNAs (miR-16, miR-25, miR-222 and miR-324-3p) were differentially expressed between breast cancer cases and controls and subsequently validated in the internal and external validation stage samples. Taken together, Chinese patients with high plasma miRNA expression levels had a significantly increased risk of breast cancer for all the four miRNAs. The Area Under the Receiver Operating Characteristic Curve was 0.917 for the four-miRNA signature in the discovery stage (sensitivity=0.917 and specificity=0.917), 0.930 in the internal validation stage (sensitivity=0.920 and specificity=0.940). Conclusions The four-miRNA signature from plasma may serve as a noninvasive diagnostic biomarker for breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 161. doi:10.1158/1538-7445.AM2011-161

Abstract 3002: Serum miRNA signatures predict survival of NSCLC

Abstract Purpose Recent findings that human serum contains stably expressed microRNAs (miRNAs) have revealed a great potential of serum miRNA signature as disease fingerprints to predict survival. We used genome-wide serum miRNA expression analysis to investigate the role of serum miRNA in predicting prognosis of NSCLC. Patients and Methods To control disease heterogeneity, we used patients with stage I-IIIa lung adenocarcinoma and squamous cell carcinoma, who were treated with both operation and adjuvant chemotherapies. In the discovery stage, Solexa sequencing followed by individual quantitative RT-PCR (qRT-PCR) assays was used to test the difference in levels of serum miRNAs between 30 longer survival patients (alive and mean survival time: 49.54 months) and 30 shorter survival patients (dead and mean survival time: 9.54 months) matched by age, sex and stage. The detected serum miRNAs were then validated in 243 patients (randomly classified into two subgroups: 120 for the training set and 123 for the testing set). Results Eleven serum miRNAs were found to be altered more than 5-fold by Solexa sequencing between “longer survival” and “shorter survival” groups, and have at least 50 copies in either “longer survival” or “shorter survival” groups. We then performed individual qRT-PCR assay to quantify each of the identified miRNAs and the levels of four miRNAs (miR-486, miR-30d, miR-1 and miR-499) were significantly different between the “longer survival” and “shorter survival” groups (P = 0.0002 for miR-486, 4.30 × 10 −7 for miR-30d, 0.001 for miR-1 and 0.0001 for miR-499). From the training set, high serum expression levels of miR-486 (median: 0.795) and miR-30d (median: 0.680), and low serum expression levels of miR-1 (median: 0.675), miR-499 (median: 0.750) were all individually associated with unfavorable survival. When these threshold values were applied to the testing set, comparable log-rank P values and HRs were also observed, respectively. Results for all the 303 samples including the discovery dataset were also similar to those from the training set. Patients with high serum expression levels of risk miRNAs and low serum expression levels of protective miRNAs had significantly lower MSTs (31.30 months for miR-486, 22.37 months for miR-30d, 22.87 months for miR-1 and 23.73 months for miR-499, all versus not reached). In the combined analysis, we found that patients carrying two or more high-risk miRNAs had significantly increased probability of cancer death in a dose-dependent manner (log rank test: P < 0.0001, HR = 3.14, 95%CI = 1.65-5.97 for 2 high-risk miRNAs carriers, HR = 16.52, 95%CI = 8.62-31.68 for 3 high-risk miRNAs carriers and HR = 34.13, 95%CI = 16.28-71.56 for 4 high-risk miRNAs carriers), compared with those carrying zero or one high-risk miRNA. Conclusion The four-miRNA signature from the serum may serve as a non-invasive predictor for the overall survival of NSCLC. 1 2 Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3002.

Serum MicroRNA Signatures Identified in a Genome-Wide Serum MicroRNA Expression Profiling Predict Survival of Non–Small-Cell Lung Cancer

Recent findings that human serum contains stably expressed microRNA (miRNA) have revealed a great potential of serum miRNA signature as disease fingerprints to predict survival. We used genome-wide serum miRNA expression analysis to investigate the role of serum miRNA in predicting prognosis of non-small-cell lung cancer (NSCLC). To control disease heterogeneity, we used patients with stages I to IIIa lung adenocarcinoma and squamous cell carcinoma, who were treated with both operation and adjuvant chemotherapies. In the discovery stage, Solexa sequencing followed by individual quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) assays was used to test the difference in levels of serum miRNAs between 30 patients with longer survival (alive and mean survival time, 49.54 months) and 30 patients with shorter survival matched by age, sex, and stage (dead and mean survival time, 9.54 months). The detected serum miRNAs then were validated in 243 patients (randomly classified into two subgroups: n = 120 for the training set, and n = 123 for the testing set). Eleven serum miRNAs were found to be altered more than five-fold by Solexa sequencing between longer-survival and shorter-survival groups, and levels of four miRNAs (ie, miR-486, miR-30d, miR-1 and miR-499) were significantly associated with overall survival. The four-miRNA signature also was consistently an independent predictor of overall survival for both training and testing samples. The four-miRNA signature from the serum may serve as a noninvasive predictor for the overall survival of NSCLC.