ccRCC Cases Research Articles

Evaluation of a seven gene mutational profile as a prognostic factor in a population-based study of clear cell renal cell carcinoma

In this study, we investigate the influence of the seven genes (VHL, PBRM1, SETD2, BAP1, KDM5C, MTOR and TP53) most frequently mutated in clear cell renal cell cancer (ccRCC) on cancer-specific survival (CSS) in the prospective Netherlands Cohort Study on diet and cancer. DNA isolated from routinely archived formalin-fixed paraffin-embedded tumour blocks from 252 incident ccRCC cases was available for targeted next generation sequencing. Based on the sequencing quality and the completeness of information on clinical characteristics and follow-up, we could use 110 cases for survival analysis. The association with CSS for each mutated gene in these cases was tested using multivariable Cox proportional hazards models to estimate hazards ratios (HR) and confidence intervals (CIs), and we observed mutations in one or more of the seven genes in 64 out of 110 cases (58%). In the multivariable-adjusted analyses, mutations in VHL and PBRM1 were associated with better CSS (HRs (95% CI) 0.34 (0.13‒0.89) and 0.17 (0.04–0.66), respectively), although these results were not statistically significant after multiple testing correction. No association was observed for the other five genes, which may be attributable to limited power.

LZTS2: A novel and independent prognostic biomarker for clear cell renal cell carcinoma

PurposeLeucine zipper tumor suppressor 2 (LZTS2), a putative tumor suppressor gene, has been demonstrated to be a negative regulator of microtubule severing during cytokinesis and a negative regulator of the Wnt signaling pathway. In a genetically modified mouse model, deletion of Lzts2 altered normal ureteric bud branching morphogenesis and caused cystogenesis in mice. Cyst-lining cells demonstrated atypical features, closely resembling those observed in mouse models of human clear cell renal cell carcinoma (ccRCC), which could represent a preneoplastic lesion. These findings suggest that LZTS2 may play a role in ccRCC tumorigenesis. The aim of this study was to establish an association between LZTS2 differential expression and clinicopathological features of ccRCC and to investigate its prognostic value as well as the underlying mechanisms in ccRCC progression. Materials and MethodsGene expression data by RNA-sequencing for cohorts of 510 ccRCC cases with clinical outcome data were extracted from The Cancer Genome Atlas (TCGA) using cBioPortal. Chi-square test of independence, Kaplan-Meier curves, and Cox regression models were used to investigate the possible relationship between LZTS2 mRNA expression levels and clinicopathological parameters as well as patient survival to establish its prognostic values. To examine its cellular localization, we performed LZTS2 antibody staining and scored the expression levels in a pilot study on a tissue microarray (TMA) containing 31 clear cell RCCs, 32 chromophobe RCCs, 12 papillary RCCs, and 20 adjacent benign renal tissue, as well as placental tissue diagnosed between 2001 and 2007 at the University of California San Francisco Medical Center. Staining was subsequently repeated in 15 ccRCCs on whole section slides to confirm the results. ResultsOur analysis of TCGA data demonstrated that LZTS2 expression levels were associated with tumor grade (p = 0.005), T stage (p < 0.001), metastasis status (p < 0.001), and overall clinical stage (p < 0.001). High level of expression was correlated with worse overall survival (p < 0.001), disease-specific survival (p < 0.001), progression-free survival (p < 0.001), and disease-free survival (p < 0.001) compared to low level of expression. Multivariate Cox regression analysis revealed that high LZTS2 expression was an independent poor prognostic factor for overall survival (HR = 2.083, p < 0.001), disease-specific survival (HR = 2.298, p < 0.001), and progression-free survival (HR = 1.896, p < 0.001) in patients with ccRCC. A few known driver mutations in ccRCC pathogenesis, including BAP1, NF2, and RELN, were enriched in LZTS2 high expression tumors. In particular, LZTS2 expression level could be a biomarker for risk stratification of the prognosis of BAP1-mutated ccRCCs. Immunohistochemical staining with anti-LZTS2 antibody was performed to examine its cellular localization in ccRCC and demonstrated centrosomal and acentrosomal distribution in tumors of various Fuhrman nuclear grades. Furthermore, high LZTS2 cytoplasmic expression was associated with centrosomal amplification (p = 0.030) in this small-scale study. ConclusionsThe current study established an independent prognostic value of LZTS2 expression in ccRCC and explored the molecular mechanisms of LZTS2 in predicting the prognosis of ccRCC. Further studies are needed to validate our analysis and to provide a precise understanding of the function of LZTS2 in ccRCC.

Associations between tumor grade, contrast-enhanced ultrasound features, and microvascular density in patients with clear cell renal cell carcinoma: a retrospective study.

Clear cell renal cell carcinoma (ccRCC) comprises 70% of all renal cell carcinomas (RCCs). Currently, the most important prognostic factor for this type of carcinoma is the World Health Organization/International Society of Urological Pathology (WHO/ISUP) grade. However, nonsurgical methods are rarely used to determine a tumor's WHO/ISUP grade, thus limiting the development of nonsurgical therapies. Due to variations in microvascular density (MVD) at different stages of tumor growth, contrast-enhanced ultrasound (CEUS) features may provide a noninvasive method for evaluating the WHO/ISUP grade of ccRCC. In this study, we analyzed confirmed cases of ccRCC using CEUS features. We also used CD34 and CD31 antibodies to determine MVD. The heterogeneity of CD34 and CD31 expressions were used to determine different degrees of angiogenesis. When compared to WHO/ISUP grade I/II (G1/G2) tumors, grade III/IV (G3/G4) tumors had reduced peak intensity (PI) (P=0.006), time to peak (TTP) (P<0.001), and relative enhancement percentage index (∆PI%) (P<0.001). However, the frequency of incomplete pseudocapsule (P=0.049) and slow wash-in (P=0.001) was significantly higher in G3/G4 tumors. A cut-off value of ∆PI% <33.15% (P<0.001) allowed identification of G3/G4 tumors with an area under the curve (AUC) of 0.80 [95% confidence interval (CI): 0.70 to 0.91) and a sensitivity of 80%. The mean CD34+ MVD (P<0.001) and CD31+ MVD (P<0.001) were significantly lower in G3/G4 tumors. A positive correlation was revealed between ∆PI% and MVD. There was a statistically significant difference in the density of undifferentiated vessels between the slow wash-in and fast wash-in cases (P<0.001). The features of CEUS are effective for differentiating G3/G4 tumors from G1/G2. There was a positive correlation detected between ∆PI% and MVD, and the density of undifferentiated vessels showed a significant difference between slow wash-in and fast wash-in cases. These findings indicate that CEUS can enable the sonographic visualization of tumor angiogenesis and thus be considered an acceptable method for the nonsurgical assessment of tumor microvascular distribution and grade.

Comprehensive genomic profiling (CGP) of chromophobe renal cell carcinoma (chrRCC) compared with clear cell RCC (ccRCC): Impact of FLCN genomic alteration (GA) status.

292 Background: FLCN is a tumor suppressor gene associated with cutaneous hair follicle development. FLCN germline mutations are linked to inherited chrRCC in the Birt-Hogg-Dube (BHD) syndrome. We queried whether clinically sporadic chrRCC featured FLCN mutations by comparing the genomic profiles of chrRCC with ccRCC. Methods: 108 chrRCC and 2110 ccRCC underwent hybrid-capture based comprehensive genomic profiling (CGP) to evaluate all classes of genomic alterations (GA). Tumor mutational burden (TMB) was determined on up to 1.1 Mbp of sequenced DNA and microsatellite instability (MSI) was determined on 114 loci. PD-L1 expression was determined by IHC (Dako 22C3). Results: Patients (pts) with chrRCC were more frequently female and younger than pts with ccRCC p &lt;.0001). None of the submitted clinical records in the chrRCC cases listed signs of BHD syndrome. FLCN GA were identified in only 2.3% of the ccRCC cases with 37% of the GA predicted to be germline. chrRCC did not reveal somatic or germline FLCN GA. GA/tumor were slightly higher in ccRCC vs chrRCC (3.6 vs 2.4; NS). GA more frequent in chrRCC included TP53, RB1 and PTEN. GA more frequent in the ccRCC included VHL, BAP1, PBRM1, SETD2, CDKN2A/B, ARID1A, NF2, PIK3CA and TERT. Putative biomarkers of immune checkpoint inhibitor (ICPI) response were infrequent in both groups with only a slightly higher, but still low, mean TMB in ccRCC vs chrRCC cases. IHC revealed moderate PD-L1 expression at low and minimal PD-L1 expression at high staining level, which was slightly increased in the chrRCC group. Conclusions: FLCN mutations that are associated with the familial incidence of chrRCC were not associated with sporadic chrRCC. Sporadic chrRCC has substantially different genomic profile from ccRCC and may harbor a few ‘targetable’ GA. The prediction of response to ICPI in RCC remains challenging with chrRCC featuring slightly higher PD-L1 expression and ccRCC featuring higher PBRM1 GA and higher TMB.[Table: see text]

Identification of UBE2I as a Novel Biomarker in ccRCC Based on a Large-Scale CRISPR-Cas9 Screening Database and Immunohistochemistry.

Background: The genome-wide CRISPR-cas9 dropout screening has emerged as an outstanding approach for characterization of driver genes of tumor growth. The present study aims to investigate core genes related to clear cell renal cell carcinoma (ccRCC) cell viability by analyzing the CRISPR-cas9 screening database DepMap, which may provide a novel target in ccRCC therapy. Methods: Candidate genes related to ccRCC cell viability by CRISPR-cas9 screening from DepMap and genes differentially expressed between ccRCC tissues and normal tissues from TCGA were overlapped. Weighted gene coexpression network analysis, pathway enrichment analysis, and protein–protein interaction network analysis were applied for the overlapped genes. The least absolute shrinkage and selection operator (LASSO) regression was used to construct a signature to predict the overall survival (OS) of ccRCC patients and validated in the International Cancer Genome Consortium (ICGC) and E-MTAB-1980 database. Core protein expression was determined using immunohistochemistry in 40 cases of ccRCC patients. Results: A total of 485 essential genes in the DepMap database were identified and overlapped with differentially expressed genes in the TCGA database, which were enriched in the cell cycle pathway. A total of four genes, including UBE2I, NCAPG, NUP93, and TOP2A, were included in the gene signature based on LASSO regression. The high-risk score of ccRCC patients showed worse OS compared with these low-risk patients in the ICGC and E-MTAB-1980 validation cohort. UBE2I was screened out as a key gene. The immunohistochemistry indicated UBE2I protein was highly expressed in ccRCC tissues, and a high-level nuclear translocation of UBE2I occurs in ccRCC. Based on the area under the curve (AUC) values, nuclear UBE2I had the best diagnostic power (AUC = 1). Meanwhile, the knockdown of UBE2I can inhibit the proliferation of ccRCC cells. Conclusion: UBE2I, identified by CRISPR-cas9 screening, was a core gene-regulating ccRCC cell viability, which accumulated in the nucleus and acted as a potential novel promising diagnostic biomarker for ccRCC patients. Blocking the nuclear translocation of UBE2I may have potential therapeutic value with ccRCC patients.

Clinicopathological profile of clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer with aggressive behavior. Tumor stage and grade are important parameter in diagnosis. Data about the clinical and pathology profile of ccRCC in Indonesia is limited, therefore further information is needed. The study aimed to determine clinicopathologic profile of ccRCC in Soetomo Academic General Hospital (SAGH). This descriptive study was conducted retrospectively with cross-sectional approach. Clinical and histopathological archives of ccRCC specimen were retrieved during January 2016 to December 2020. The total number of ccRCC cases obtained was 38 cases, the most cases (11) occurred in 2016. The average of patient’s age was 55.64 years. Patients were predominantly male (71.1%), with a ratio to female 2.5:1. The tumors mostly filled the entire kidney, in 15 (39.5%) cases. Most pT stage distribution was pT2, in 15 (39.5%) cases. Most N stage distribution was N0, in 23 (60.5%) cases. Only 5 (13.2%) cases showed multiple pulmonary nodules which suspected to be metastatic (M1). The most common tumors grade was G4, in 17 (44.7%) cases. In conclusion, ccRCC patient were predominantly male, tumors mostly high grade, filled entire kidney, not penetrated renal capsule, and not metastasize to regional lymph nodes.

Overexpression of PFKFB3 promotes cell glycolysis and proliferation in renal cell carcinoma

BackgroundCancer cells prefer utilizing aerobic glycolysis in order to exacerbate tumor mass and maintain un-regulated proliferative rates. As a key glycolytic activator, phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) has been implicated in multiple tumor type progression. However, the specific function and clinical significance of PFKFB3 in renal cell carcinoma (RCC) are yet not clarified. This investigation assessed PFKFB3 roles in RCC.MethodsPFKFB3 expression levels were analyzed in clear cell renal cell carcinoma (ccRCC) tissues, together with its relationship with clinical characteristics of ccRCC. Real-time PCR and Western blot assays were employed for determining PFKFB3 expression in different RCC cell lines. Furthermore, we determined the glycolytic activity by glucose uptake, lactate secretion assay and ECAR analysis. CCK-8 assay, clone formation, flow cytometry and EdU assessments were performed for monitoring tumor proliferative capacity and cell-cycle distribution. Furthermore, a murine xenograft model was employed for investigating the effect of PFKFB3 on tumor growth in vivo.ResultsPFKFB3 was significantly up-regulated in RCC specimens and cell lines in comparison to normal control. Overexpression of PFKFB3 was directly correlated to later TNM stages, thus becoming a robust prognostic biomarker for ccRCC cases. Furthermore, PFKFB3 knockdown suppressed cell glycolysis, proliferative rate and cell-cycle G1/S conversion in RCC cells. Importantly, in vivo experiments confirmed that PFKFB3 knockdown delayed tumor growth derived from the ACHN cell line.ConclusionsSuch results suggest that PFKFB3 is a key molecular player in RCC progression via mediating glycolysis / proliferation and provides a potential therapeutic target against RCC.

Efficacy of nivolumab plus ipilimumab as first-line therapy for primary tumors in patients with renal cell carcinoma.

With the emergence of several effective combination therapies, information on their effects at the primary site will be crucial for planning future cytoreductive nephrectomy (CN). The present study focused exclusively on changes in primary tumor sizes following treatment with nivolumab plus ipilimumab and investigated the clinical factors associated with a good response in primary tumors. We retrospectively assessed 27 patients diagnosed with advanced renal cell carcinoma (RCC) who started treatment with nivolumab plus ipilimumab. Changes in tumor sizes at the primary site were described using waterfall and spider plots, respectively. We analyzed the correlation of tumor shrinkage between primary and metastatic site. The parameters analyzed between responders and non-responders according to primary tumor sizes were International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) risk scores, peripheral blood markers, and CRP. The median age and follow-up period were 66 years and 9.3 months, respectively. The median IMDC risk score was 3 (range: 1-6). Nineteen patients were diagnosed with clear-cell RCC (ccRCC) and 8 patients with non-ccRCC. Among ccRCC patients, 9 (47.4%) achieved a significant response with a maximum reduction of 30% or more in the size of the primary tumor from baseline within 4 months, while 3 (37.5%) out of 8 patients with non-ccRCC achieved a significant response. Shrinkage of the primary tumor correlated with the metastatic tumors in both ccRCC and non-ccRCC cases. Of note, 6 patients underwent CN and no viable tumor cells were detected in the surgical specimens of 3 patients whose primary tumors shrank by approximately 50%-60% with a reduction to 4 cm or less. Among ccRCC patients, the neutrophil-to-lymphocyte ratio and monocyte-to-lymphocyte ratio were slightly lower in responders than in non-responders (P = 0.0944 and P = 0.0691). The platelet-to-lymphocyte ratio was significantly lower in responders than in non-responder (P = 0.0391). Significant responses in primary tumors to nivolumab plus ipilimumab were observed in 50% of ccRCC patients, while responses varied among non-ccRCC patients. Inflammation markers may be predictive factors of treatment responses in primary tumors. Although further studies are needed, the present results suggest the importance of considering CN from radiological and pathological viewpoints.

Heterogeneity of Platelet Derived Growth Factor Pathway Gene Expression Profile Defines Three Distinct Subgroups of Renal Cell Carcinomas.

We previously described four different vascular patterns (reticular, diffuse, fasciculate, and trabecular) in renal cell carcinoma (RCC) suggesting an early and heterogeneous acquisition of perivascular cells most probably due to a particular PDGF pathway gene expression profile. The aim of the study was to study PDGF pathway gene expression profiles, separately for each vascular pattern. TaqMan assay for the PDGF pathway was performed on twelve cases of ccRCC previously evaluated by histopathology, immunohistochemistry, and RNAscope. Gene expression profile was correlated with grade, invasion, vascular patterns, and VEGF. PIK3C3 and SLC9A3 genes were overexpressed in all vascular patterns, but they were significantly correlated with high VEGF mRNA in the reticular and diffuse pattern. STAT1, JAK2, SHC2, SRF and CHUK (IKK) were exclusively overexpressed in cases with diffuse vascular pattern. SLC9A3, CHUK and STAT3 were overexpressed in G2 tumors. Three ccRCC subgroups were defined: 1) PIK3C3 (VSP34)/SLC9A3 which may be proper for anti PIK3C3 inhibitors; 2) VEGFhigh subgroup where association of anti VEGF may be a benefit and 3) JAK2/STAT1 subgroup, potentially being eligible for anti JAK/STAT therapy associated with IKK inhibitors.

Prostate-specific membrane antigen expression in clear-cell renal cell carcinoma

Background Clear-cell renal cell carcinoma (ccRCC) represents the majority of renal neoplasms with usual late presentation. Metastasis of ccRCC is very common owing to the vascular nature of this tumor. Identifying markers that could predict tumor behavior and metastatic potential is crucial to improve patient prognosis. Prostate-specific membrane antigen (PSMA) is a recently identified vascular marker that is expressed in many cancer types. This study aimed at evaluation of its clinicopathologic role in ccRCC. Materials and methods A total of 40 cases of ccRCC were recruited and subjected to immunohistochemical staining for PSMA. Statistical analysis was performed to assess its expression and its relationship with different clinicopathologic parameters. Results PSMA expression was positive in 72.5% of all cases. High expression was detected in 52.5% of cases, with positive correlation with high tumor grade (P=0.01), stage (0.04), and lymphovascular invasion (P=0.03). PSMA expression was positive in 83.3% of cases of tumor thrombus tissue, and 50% of these cases showed high expression. Two (50%) cases of metastatic lesions showed low positive expression for PSMA with high PSMA expression in 75% of the corresponding primary lesions. Conclusion PSMA is specifically expressed in the vasculature of ccRCC, showing a positive correlation with poor prognostic parameters. This renders PSMA as a promising prognostic marker and therapeutic target.

Enhanced computed tomography radiomics-based machine-learning methods for predicting the Fuhrman grades of renal clear cell carcinoma.

To develop and test an optimal machine learning model based on the enhanced computed tomography (CT) to preoperatively predict pathological grade of clear cell renal cell carcinoma (ccRCC). A retrospective analysis of 53 pathologically confirmed cases of ccRCC was performed and 25 consecutive ccRCC cases were selected as a prospective testing set. All patients underwent routine preoperative abdominal CT plain and enhanced scans. Renal tumor lesions were segmented on arterial phase images and 396 radiomics features were extracted. In the training set, seven discrimination classifiers for high- and low-grade ccRCCs were constructed based on seven different machine learning models, respectively, and their performance and stability for predicting ccRCC grades were evaluated through receiver operating characteristic (ROC) analysis and cross-validation. Prediction accuracy and area under ROC curve were used as evaluation indices. Finally, the diagnostic efficacy of the optimal model was verified in the testing set. The accuracies and AUC values achieved by support vector machine with radial basis function kernel (svmRadial), random forest and naïve Bayesian models were 0.860±0.158 and 0.919±0.118, 0.840±0.160 and 0.915±0.138, 0.839±0.147 and 0.921±0.133, respectively, which showed high predictive performance, whereas K-nearest neighborhood model yielded lower accuracy of 0.720±0.188 and lower AUC value of 0.810±0.150. Additionally, svmRadial had smallest relative standard deviation (RSD, 0.13 for AUC, 0.17 for accuracy), which indicates higher stability. svmRadial performs best in predicting pathological grades of ccRCC using radiomics features computed from the preoperative CT images, and thus may have high clinical potential in guiding preoperative decision.

Involvement of PBRM1 in VHL disease-associated clear cell renal cell carcinoma and its putative relationship with the HIF pathway.

Von Hippel-Lindau (VHL) disease is the main cause of inherited clear-cell renal cell carcinoma (ccRCC) and is caused by germline mutations in the VHL tumor suppressor gene. Bi-allelic VHL alterations lead to inactivation of pVHL, which plays a major role by downstream activation of the hypoxia inducible factor (HIF) pathway. Somatic VHL mutations occur in 80% of sporadic ccRCC cases and the second most frequently mutated gene is polybromo 1 (PBRM1). As there is currently no data regarding PBRM1 involvement in VHL disease-associated ccRCC, the aim of the present study was to assess the PBRM1 mutational status, and PBRM1 and HIF expression in VHL disease-associated ccRCC series compared with a sporadic series. PBRM1 gene was screened by Sanger sequencing for 23 VHL-disease-associated ccRCC and 22 sporadic ccRCC cases. Immunohistochemical studies were performed to detect the expression of PBRM1, HIF1 and HIF2 for all cases. In VHL-associated tumors, 13.0% (n=3/23) had PBRM1 somatic mutations and 17.4% (n=4/23) had a loss of PBRM1 nuclear expression. In sporadic cases, 27.3% (n=6/22) showed PBRM1 somatic mutations and 45.5% (n=10/22) had a loss of PBRM1 nuclear expression. Loss of PBRM1 was associated with an advanced tumor stage. HIF1-positive tumors were observed more frequently in the VHL-associated ccRCC than in the sporadic series. Furthermore, in the VHL cohort, PBRM1 expression appeared to be associated more with HIF1 than with HIF2. Given that hereditary tumors tend to be less aggressive, these results would suggest that co-expression of PBRM1 and HIF1 may have a less oncogenic role in VHL-associated ccRCC.

Prognostic Value of Metabolism-Related Genes and Immune Infiltration in Clear Cell Renal Cell Carcinoma.

BackgroundClear cell renal cell carcinoma (ccRCC) is one of the most prevalent cancers. Thus, it is warranted to detect the status of metabolism-related genes (MRGs) and infiltrating immune cells in ccRCC progression for the prognosis of ccRCC. This research was designed to establish and verify the prognostic signature of ccRCC using MRGs. In addition, we investigated the potential link between the relative proportion of tumor infiltrated immune cells (TIICs) and ccRCC prognosis.MethodsSequencing data of metabolism-related gene sets in ccRCC cases were obtained from The Cancer Genome Atlas database (TCGA) and Gene Expression Omnibus Database (GEO). The R Programming Language software packages were applied for differential analysis of MRGs. First, a univariate Cox regression model was applied to determine the MRGs linked with overall survival (OS). Then, the multivariate Cox regression model was applied to establish the prognostic signature. Finally, the CIBERSORT algorithm was used to determine the proportion of TIICs.ResultsOverall, 286 differentially expressed MRGs were identified in the TCGA dataset. Univariate and multivariate Cox regression models were applied to develop a prognostic signature with six MRGs. The predictive capability of the prognostic signature was further verified by TCGA and GEO database. In addition, RS positively correlated with memory B cells, plasma cells, activated memory CD4+ T cells, follicular helper T cells, regulatory T cells, CD8+ T cells, and M0 macrophages, and were negatively associated with resting memory CD4+ T cells, resting dendritic cells, activated dendritic cells, M2 macrophages, monocytes, resting mast cells, and eosinophils.ConclusionHerein, a prognostic signature was developed using MRGs for ccRCC prognosis. The proportion of 22 TIICs in ccRCC and the association between TIICs and clinical outcomes were also determined. The identified genes and cells could guide future targeted therapy and immunotherapy.

Identification and validation of a hypoxia-related prognostic signature in clear cell renal cell carcinoma patients.

Increasing evidence has shown that hypoxia is closely related to the development, progression, and prognosis of clear cell renal cell carcinoma (ccRCC). Nevertheless, reliable prognostic signatures based on hypoxia have not been well-established. This study aimed to establish a hypoxia-related prognostic signature and construct an optimized nomogram for patients with ccRCC.We accessed hallmark gene sets of hypoxia, including 200 genes, and an original RNA seq dataset of ccRCC cases with integrated clinical information obtained by mining the Cancer Genome Atlas database and the International Cancer Genome Consortium (ICGC) database. Univariate Cox regression analysis and multivariate Cox proportional hazards regression were performed to identify prognostic hub genes and further established prognostic model as well as visualized the nomogram. External validation of the optimized nomogram was performed in independent cohorts from the ICGC database.ANKZF1, ETS1, PLAUR, SERPINE1, FBP1, and PFKP were selected as prognostic hypoxia-related hub genes, and the prognostic model effectively distinguishes high-risk and low-risk patients with ccRCC. The results of receiver operating characteristic curve, risk plots, survival analysis, and independent analysis suggested that RiskScore was a useful tool and independent predictive factor. A novel prognosis nomogram optimized via RiskScore showed its promising performance in both the Cancer Genome Atlas-ccRCC cohort and an ICGC-ccRCC cohort.Our study reveals that the differential expressions of hypoxia-related genes are associated with the overall survival of patients with ccRCC. The prognostic model we established showed a good predictive and discerning ability in ccRCC patients. The novel nomogram optimized via RiskScore exhibited a promising predictive ability. It may be able to serve as a visualized tool for guiding clinical decisions and selecting effective individualized treatments.

Characterization of Molecular Heterogeneity Associated With Tumor Microenvironment in Clear Cell Renal Cell Carcinoma to Aid Immunotherapy.

Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer and has strong immunogenicity. A systematically investigation of the tumor microenvironment (TME) in ccRCC could contribute to help clinicians develop personalized treatment and facilitate clinical decision-making. In this study, we analyzed the immune-related subtype of ccRCC on the basis of immune-related gene expression data in The Cancer Genome Atlas (TCGA, N = 512) and E-MTAB-1980 (N = 101) dataset, respectively. As a result, two subtypes (C1 and C2) were identified by performing non-negative matrix factorization clustering. Subtype C1 was characterized by increased advance ccRCC cases and immune-related pathways. A higher immune score, stromal score, TMB value, Tumor Immune Dysfunction and Exclusion (TIDE) prediction score, and immune checkpoint genes expression level were also observed in C1. In addition, the C1 subtype might benefit from chemotherapy and immunotherapy. The patients in subtype C2 had more metabolism-related pathways, higher tumor purity, and a better prognosis. Moreover, some small molecular compounds for the treatment of ccRCC were identified between the two subtypes by using the Connectivity Map (CMap) database. Finally, we constructed and validated an immune-related (IR) score to evaluate immune modification individually. A high IR score corresponded to a favorable prognosis compared to a low IR score, while more advanced tumor stage and grade cases were enriched in the low IR score group. The two IR score groups also showed a distinct divergence among immune status, TME, and chemotherapy. The external validation dataset (E-MTAB-1980) and another immunotherapy cohort (IMvigor 210) demonstrated that patients in the high IR score group had a significantly prolonged survival time and clinical benefits compared to the low IR score group. Together, characterization of molecular heterogeneity and IR signature may help develop new insights into the TME of ccRCC and provide new strategies for personalized treatment.

Eosinophilic features in clear cell renal cell carcinoma correlate with outcomes of immune checkpoint and angiogenesis blockade

BackgroundClear cell renal cell carcinoma (ccRCC) displays heterogeneity in appearance—a distinctive pale clear to eosinophilic cytoplasm; however, little is known about the underlying mechanisms and clinical implications. We investigated the...

MP14-16 EFFICACY OF NIVOLUMAB PLUS IPILIMUMAB IN PRIMARY TUMOR AS FIRST-LINE THERAPY IN THE PATIENTS WITH ADVANCED RENAL CELL CARCINOMA

You have accessJournal of UrologyKidney Cancer: Advanced (including Drug Therapy) I (MP14)1 Sep 2021MP14-16 EFFICACY OF NIVOLUMAB PLUS IPILIMUMAB IN PRIMARY TUMOR AS FIRST-LINE THERAPY IN THE PATIENTS WITH ADVANCED RENAL CELL CARCINOMA Hiroshi Kikuchi, Takahiro Osawa, Ryuji Matsumoto, Takashige Abe, Satoru Maruyama, Toru Harabayashi, Haruka Miyata, Akira Kashiwagi, Ataru Sazawa, Riyo Fukui, Ken Morita, Ichiro Takeuchi, Noboru Yamashita, Keita Minami, Tango Mochizuki, and Nobuo Shinohara Hiroshi KikuchiHiroshi Kikuchi More articles by this author , Takahiro OsawaTakahiro Osawa More articles by this author , Ryuji MatsumotoRyuji Matsumoto More articles by this author , Takashige AbeTakashige Abe More articles by this author , Satoru MaruyamaSatoru Maruyama More articles by this author , Toru HarabayashiToru Harabayashi More articles by this author , Haruka MiyataHaruka Miyata More articles by this author , Akira KashiwagiAkira Kashiwagi More articles by this author , Ataru SazawaAtaru Sazawa More articles by this author , Riyo FukuiRiyo Fukui More articles by this author , Ken MoritaKen Morita More articles by this author , Ichiro TakeuchiIchiro Takeuchi More articles by this author , Noboru YamashitaNoboru Yamashita More articles by this author , Keita MinamiKeita Minami More articles by this author , Tango MochizukiTango Mochizuki More articles by this author , and Nobuo ShinoharaNobuo Shinohara More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000001995.16AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: After the findings from CARMENA study, the role of cytoreductive nephrectomy (CN) should be considered in patient-tailored manner. Taking together with the new emergences of several effective combination therapies, the information of those effect on the primary site should be crucial for planning a future CN. This study aimed to focus exclusively on the size change of primary tumor following nivolumab plus ipilimumab (NivoIpi) treatment and elucidate the clinical factors associated with good response in primary tumor. METHODS: We retrospectively assessed 27 patients who were diagnosed with advanced renal cell carcinoma (RCC) and who had started NivoIpi. The changes in tumor size of the primary site were described using waterfall and spider plots. The parameters analyzed between responder and non-responder in the size of the primary tumor were International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) risk scores, peripheral blood markers and CRP. The ANOVA test was applied for the statistical analysis. RESULTS: The median age and follow-up periods were 66 years and 8.8 months, respectively. Nineteen patients were diagnosed with clear-cell renal cell carcinoma (ccRCC). Figure 1Ashow the maximum reduction from baseline in primary tumor and best overall response in ccRCC patients. Among ccRCC patients, 9 (47.4%) patients demonstrated significant response with 30% or more reduction from baseline within 4 months in primary tumors (Figure 1B). Of 6 patients who underwent nephrectomy, 3 patients whose primary tumors shrinked around 50-60% with reduction to 4 cm or less revealed no viable tumor cell in surgical specimens. Neutrophil-to-lymphocyte ratio and monocyte-to-lymphocyte ratio tended to decrease in those responders compare to non-responders among ccRCC patients (P=0.0944 and P=0.0691). Platelet-to-lymphocyte ratio significantly decreased in those responders (P=0.0391). CONCLUSIONS: In our experience, significant responses of primary tumor were obtained with NivoIpi in half cases of ccRCC. Of note, inflammation markers were possible predictive factor for treatment response of primary tumor. Although further study is needed, these observations help us to consider CN from the viewpoint of the radiological and pathological aspects. Source of Funding: None declared © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e259-e259 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hiroshi Kikuchi More articles by this author Takahiro Osawa More articles by this author Ryuji Matsumoto More articles by this author Takashige Abe More articles by this author Satoru Maruyama More articles by this author Toru Harabayashi More articles by this author Haruka Miyata More articles by this author Akira Kashiwagi More articles by this author Ataru Sazawa More articles by this author Riyo Fukui More articles by this author Ken Morita More articles by this author Ichiro Takeuchi More articles by this author Noboru Yamashita More articles by this author Keita Minami More articles by this author Tango Mochizuki More articles by this author Nobuo Shinohara More articles by this author Expand All Advertisement Loading ...

High Immunoexpression of COX-2 as a Metastatic Risk Factor in ccRCC without PD-L1 Involvement.

IntroductionClear cell renal cell carcinoma (ccRCC) is the most lethal type of malignancy of the urinary tract system as it is resistant to chemotherapy and radiation and has a survival rate of less than 5% in cases of metastasis. Inflammation plays an essential role in the metastasis of ccRCC. Cyclooxygenase-2 (COX-2) is an inflammatory protein that affects the processes of carcinogenesis, invasion, migration, metastasis, and angiogenesis. COX-2 can modulate programmed death ligand-1 (PD-L1) expression and play a role in immune evasion, meaning that tumor cells are able to escape the body’s immune response and more easily metastasize.PurposeThis study aims to determine the role of COX-2 and PD-L1 in the occurrence of ccRCC metastases.Materials and MethodsThis study is an observational analytical study, which employed a cross-sectional approach to examine the paraffin block samples of 40 ccRCC cases from Dr. Hasan Sadikin Hospital Bandung, Indonesia, between 2014 and 2021. Immunoexpression was measured using immunohistochemical staining for COX-2 in tumor cells and for PD-L1 in immune cells. PD-L1 calculation was measured using Qupath 0.2.3. digital software. Metastatic data were obtained using radiological imaging and pathological examinations. Meanwhile, the data were analyzed using the chi-square test for COX-2 and Fischer’s exact test for PD-L1.ResultsThe research results revealed a significant association between COX-2 and the occurrence of metastases in ccRCC (p=0.001) with a prevalence odds ratio of 10.28. Positive PD-L1 immunoexpression of the immune cells (≥1%) was found in 14% (3/21) of the metastatic group and 5% (1/19) of the non-metastatic group (p=0.607). There was no association between COX-2 and PD-L1 immunoexpression (p=0.278).ConclusionThis study shows that metastases in ccRCC patients are ten times as likely to be associated with high COX-2 immunoexpression than low COX-2 immunoexpression. COX-2 plays a role in the process of ccRCC metastasis without PD-L1 involvement.

Endothelial Reprogramming Stimulated by Oncostatin M Promotes Inflammation and Tumorigenesis in VHL-Deficient Kidney Tissue.

Clear-cell renal cell carcinoma (ccRCC) is the most prevalent subtype of renal cell carcinoma (RCC), and its progression has been linked to chronic inflammation. About 70% of the ccRCC cases are associated with inactivation of the von Hippel-Lindau (VHL) tumor-suppressor gene. However, it is still not clear how mutations in VHL, encoding the substrate-recognition subunit of an E3 ubiquitin ligase that targets the alpha subunit of hypoxia-inducible factor-α (HIFα), can coordinate tissue inflammation and tumorigenesis. We previously generated mice with conditional Vhlh knockout in kidney tubules, which resulted in severe inflammation and fibrosis in addition to hyperplasia and the appearance of transformed clear cells. Interestingly, the endothelial cells (EC), although not subject to genetic manipulation, nonetheless showed profound changes in gene expression that suggest a role in promoting inflammation and tumorigenesis. Oncostatin M (OSM) mediated the interaction between VHL-deficient renal tubule cells and the ECs, and the activated ECs in turn induced macrophage recruitment and polarization. The OSM-dependent microenvironment also promoted metastasis of exogenous tumors. Thus, OSM signaling initiates reconstitution of an inflammatory and tumorigenic microenvironment by VHL-deficient renal tubule cells, which plays a critical role in ccRCC initiation and progression. SIGNIFICANCE: A novel mechanism of cross-talk between ECs and VHL-deficient kidney tubules that stimulates inflammation and tumorigenesis is discovered, suggesting OSM could be a potential target for ccRCC intervention.

Radiomics Analysis of Contrast-Enhanced CT Predicts Survival in Clear Cell Renal Cell Carcinoma.

PurposeTo develop and validate the radiomics nomogram that combines clinical factors and radiomics features to estimate overall survival (OS) in patients with clear cell renal cell carcinoma (ccRCC), and assess the incremental value of radiomics for OS estimation.Materials and MethodsOne hundred ninety-four ccRCC cases were included in the training cohort and 188 ccRCC patients from another hospital as the test cohort. Three-dimensional region-of-interest segmentation was manually segmented on multiphasic contrast-enhanced abdominal CT images. Radiomics score (Rad-score) was calculated from a formula generated via least absolute shrinkage and selection operator (LASSO) Cox regression, after which the association between the Rad-score and OS was explored. The radiomics nomogram (clinical factors + Rad-score) was developed to demonstrate the incremental value of the Rad-score to the clinical nomogram for individualized OS estimation, which was then evaluated in relation to calibration and discrimination.ResultsRad-score, calculated using a linear combination of the 11 screened features multiplied by their respective LASSO Cox coefficients, was significantly associated with OS. Calibration curves showed good agreement between the OS predicted by the nomograms and observed outcomes. The radiomics nomogram presented higher discrimination capability compared to clinical nomogram in the training (C-index: 0.884; 95% CI: 0.808–0.940 vs. 0.803; 95% CI: 0.705–0.899, P < 0.05) and test cohorts (C-index: 0.859; 95% CI: 0.800–0.921 vs. 0.846; 95% CI: 0.777–0.915, P < 0.05).ConclusionsThe radiomics nomogram may be used for predicting OS in patients with ccRCC, and radiomics is useful to assist quantitative and personalized treatment.