Junlong Wu

Identification and validation of an eight-gene expression signature for predicting high Fuhrman grade renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is a malignancy with heterogeneous outcomes. Currently, renal mass biopsies are commonly employed to extract disease characteristics and aid prognosis. Although the pathological diagnosis of malignant disease is accurate in contemporary reports, the classification of Fuhrman grade using biopsy specimens remains far from promising. To generate a gene signature to distinguish high‐grade ccRCC, we used the cancer genome atlas (TCGA) database to develop a gene expression signature for distinguishing high‐grade (G3/4) from low‐grade (G1/2) disease. The expression profile was further validated for performance and clinical use in 283 frozen renal cancer samples and 127 ex vivo renal mass biopsy samples, respectively. The area under curve (AUC) was used to quantify discriminative ability and was compared using the De‐long test. Using the discovery dataset, we identified a 24‐gene signature for high‐grade disease with an AUC of 0.884. After applied to the development dataset, an eight‐gene profile was defined and achieved an AUC of 0.823. Accuracy of eight‐gene panel was maintained in the renal mass biopsies (RMB) samples (AUC = 0.821). In summary, using three‐stage design, we validated an eight‐gene expression signature for predicting high Fuhrman grade of ccRCC. This tool may help to reveal the characteristics of ccRCC biopsy specimens.

Low TIM3 expression indicates poor prognosis of metastatic prostate cancer and acts as an independent predictor of castration resistant status

T cell immunoglobulin 3 (TIM3) is a cell surface star molecule expressed on T cells, and also marks dysfunctional CD8+ T cells in various kinds of cancers. However, there are few studies focusing on the expression of TIM3 in tumor cells. In our study, we recruited 139 patients with metastatic prostate cancer (mPCa) who received transurethral resection of the prostate (TURP) consecutively to examine whether TIM3 expression level is associated with overall survival (OS) in mPCa patients. Immunohistochemistry was performed to determine TIM3 expression in prostate cancer tissues and then patients were divided into two groups. In multivariate Cox analysis, we revealed that mPCa patients with negative TIM3 expression, younger age, no radiotherapy, higher Gleason score, higher cT stage and patients of mCRPC had a shorter OS. Therefore, a predictive nomogram was generated with identified independent prognostic factors to assess patients’ OS at 3 years. Multivariate logistic regression revealed that higher cT stage, higher Gleason score and low TIM3 expression were independent predictors of metastatic castration resistant prostate cancer (mCRPC). In conclusion, low expression level of TIM3 in prostate cancer tissues is an independent prognostic factor of poor prognosis for mPCa patients, and also an independent predictor of mCRPC.

Renal cell carcinoma histological subtype distribution differs by age, gender, and tumor size in coastal Chinese patients

The distribution pattern of renal cell carcinoma (RCC) histological subtypes according to age, gender and tumor size has not been well illustrated in RCC patients living in fast-developing regions of China. We recruited 2941 patients with clear cell renal cell carcinoma (ccRCC), papillary renal cell carcinoma (PCC) or chromophobe from two hospitals in coastal China (2004−2012) consecutively and draw 538 American Chinese RCC patients’ data with time matched from the Surveillance, Epidemiology, and End Results database. We found that compared with ccRCC patients, chromophobe patients were more likely to be female (OR: 2.538, 95% CI: 1.923−3.350), younger (OR for 51−60 years old: 0.686; OR for over 60 years old: 0.478; reference: age < 50) and to have a larger maximal diameter (Dmax) (OR for Dmax > 7 cm: 1.883; reference: Dmax ≤ 4 cm). Besides, in comparison with coastal Chinese patients, American Chinese individuals had lower Fuhrman grades (P < 0.001) and had an onset age 10 years delay. In conclusion, we were the first to observe marked gender, age and tumor size differences in the proportional subtype distribution of RCCs in coastal Chinese patients, and also the first to compare coastal Chinese with American Chinese data.

SOX2 and SOX12 are predictive of prognosis in patients with clear cell renal cell carcinoma

Sex-determining region Y-box protein (SOX) genes serve an important role in cancer growth and metastasis. The present study aimed to determine the predictive ability of SOX and associated genes identified through molecular network in clear cell renal cell carcinoma (RCC). A total of 505 patients with clear cell RCC from The Cancer Genome Atlas (TCGA) cohorts were collected in this study. The expression profile of SOX and associated genes were obtained from the TCGA RNAseq database. Clinicopathological characteristics, including age, gender, tumor grade, stage, laterality disease-free-survival and overall survival (OS) were collected. Coxs proportional hazards regression model, as well as Kaplan-Meier curves were used to assess the relative factors. Selected genes of SOXs that demonstrated significant associations with OS were further validated in 192 patients from the validation cohort. In the univariate Cox regression model, SOX1, SOX2, SOX6, SOX11, SOX12, SOX13, SOX15, SOX17 and SOX30 expression were predictive in the prognosis of clear cell RCC. Following adjustment for clinical factors, SOX2 [hazard ratio (HR), 1.130; 95% confidence interval (CI), 1.002-1.275), SOX12 (HR, 1.379; 95% CI, 1.060-1.793) and SOX15 (HR, 1.245; 95% CI, 1.063-1.459) remained statistically significant. Furthermore, POU class 5 homeobox 1 (POU5F1), POU2F1 and nuclear receptor subfamily 5 group A member 1 in the gene cluster network analysis associated with SOX2 did not reduce the statistical significance when added to the multivariate analysis. The findings were extended to the Fudan University Shanghai Cancer Center cohort. The results revealed that high SOX2 and SOX12 expression were associated with poor prognosis for OS (log-rank test, all P<0.05). SOX2 and SOX12 were identified as independent prognostic factors of OS in clear cell RCC.

Comprehensive Analysis of BAP1 Somatic Mutation in Clear Cell Renal Cell Carcinoma to Explore Potential Mechanisms in Silico

Purpose: Aim of this study was to comprehensively analyze BRCA1-associated protein-1 (BAP1) somatic mutation in clear cell renal cell carcinoma (ccRCC) and explore potential therapeutic pathways and molecules. Patients and methods: In this study, we analyzed 445 ccRCC cases from The Cancer Genome Atlas (TCGA). Comprehensive analysis including survival, transcriptome and methylation between BAP1 mutated and wild-type cases was performed using bioinformatics tools in silico. Pathways and molecules related to BAP1 mutation were analyzed using Database for Annotation, Visualization and Integrated Discovery (DAVID) and protein-protein interaction (PPI) network. Results: BAP1 mutated ccRCC patients had a worse overall survival (OS) and disease free survival (DFS) than BAP1 wild-type patients. We found 583 up-regulated and 1216 down-regulated different expressed genes (DEGs) in BAP1 mutated tumors. Up-regulated DEGs were enriched in molecular functions and biological processes like protein binding, protein transport and ubiquitin protein ligase binding. Down-regulated DEGs were enriched in pathways like Rap1 signaling pathway, Notch pathway and altered molecular functions like metal ion binding and ubiquitin-protein transferase activity. Furthermore, CAD, TSPO, CTNNB1 and MAPK3 were top hub genes selected using PPI network analysis. Finally, BAP1 mutation had a strong correlation with CpG island methylator phenotype (CIMP). Conclusion: Our study provides a comprehensive understanding of BAP1 functional somatic mutation in ccRCC patients. Several hub genes like CAD and TSPO may become potential therapeutic targets.

MP60-13 LOSS OF FAT MASS AND OBESITY ASSOCIATED GENE (FTO) DECREASES PROLIFERATION AND INVASION IN RENAL CELL CARCINOMA CELL-LINE BY UPREGULATION OF NF-KAPPA B SIGNALING PATHWAY

INTRODUCTION AND OBJECTIVES: FTO was the first GWAS identified gene which was associated with obesity, considering the interaction between obesity and renal cell carcinoma(RCC), the biological role of FTO in RCC is of great interest. METHODS: We examined the expression of the FTO gene in RCC cell lines and the effect of FTO on cell proliferation and invasion in RCC A498 and 786-O cells. Microarray processing and analysis was used to explore novel pathways involved in FTO tumorigenesis. RESULTS: TO was expressed at high levels in A498 cells and low levels in 786-O cells. Lentivirus-mediated FTO knockdown in RCC A498 cells caused cell-cycle increase in the G1/S phase and dramatically inhibited proliferation and invasion in culture. By contrast, overexpression of FTO in 786-O cells increased proliferation and invasion. Pathway gene chip analysis revealed that FTO knockdown resulted in the NF-kappa B signaling pathway activation. IL-1R, TLR4, BIRC2, RIG-I were significantly upregulated after FTO knockdown. CONCLUSIONS: These findings demonstrate that FTO may alter proliferation, invasion and cell cycle of renal cell carcinoma cell lines through the NF-kappa B signaling pathway.

Early skeletal muscle loss during target therapy is a prognostic biomarker in metastatic renal cell carcinoma patients

Skeletal muscle depletion is common in patients with advanced cancer and may be associated with a poor outcome. To investigate whether the changes in skeletal muscle in metastatic renal cell carcinoma (mRCC) patients receiving targeted therapy are associated with clinical outcome, we undertook an observational cohort study using data from a number of randomized clinical trials previously conducted at the Fudan University Shanghai Cancer Center. The muscle mass was evaluated by comparing computed tomography images obtained at baseline with those obtained after 3–4 months of treatment. A total 101 patients were included in the study. The mean skeletal muscle area reduced from 41.6 cm2/m2 to 39.9 cm2/m2 after 3–4 months of targeted therapy. In multivariable analyses adjusted for the number of baseline covariates, muscle loss ≥5% was shown to be a significant prognostic factor for both progression-free (hazard ratio [HR]: 1.744, 95% confidence interval [CI]: 1.077–2.826, P = 0.024) and overall survival (HR: 2.367, 95%CI: 1.253–4.469, P = 0.008). The addition of muscle loss to the Heng model significantly improved its discriminative ability. In summary, early skeletal muscle loss is frequently observed in mRCC patients and can add prognostic information to current clinical risk scores.

NR1H3 expression is a prognostic factor of overall survival for patients with muscle-invasive bladder cancer

Background: Nuclear receptors (NRs) are a class of transcription factors that regulate many cellular functions through manipulation of gene expression and also play important roles in tumorigenesis, proliferation, progression and prognosis in various kinds of cancers according to recent studies. This work aimed to determine the predictive ability of NRs in muscle-invasive bladder cancer (MIBC). Patients and methods: A total of 308 MIBC patients with complete clinicopathological and RNASeq data from The Cancer Genome Atlas (TCGA) cohort were collected for filtration. Genes showed clear correlations with overall survival (OS) and recurrence free survival (RFS) were further validated in 123 MIBC patients recruited consecutively from 2008 to 2012 in Fudan University Shanghai Cancer Center (FUSCC) cohort. Cox proportional hazards regression model and Kaplan-Meier plot were used to assess the relative factors. Results: In TCGA cohort, we found that high NR1H3 (HR=0.779, 95% CI: 0.634 - 0.957), NR2C1 (HR=0.673, 95% CI: 0.458 - 0.989) and NR2F6 (HR=0.750, 95% CI: 0.574 - 0.980) expressions were independent factors of favorable OS, while only low NR1H3 (log-rank test, P=0.0076) and NR2F6 (log-rank test, P=0.0395) expressions had the ability to predict poor prognosis for RFS. Further, in FUSCC validating cohort, we confirmed that low NR1H3 expression level was independent factor of poor OS (HR=1.295, 95% CI: 1.064 - 1.576) and it had the ability to predict poor RFS (log-rank test, P=0.0059). Conclusions: Low NR1H3 expression level is an independent prognostic factor of poor OS, and can also predict worse RFS in MIBC patients. Our “TCGA filtrating and local database validating” model can help reveal more prognostic biomarkers and cast a new light in understanding certain gene function in MIBC.