Prognostic Significance of Grade Discrepancy Between Primary Tumor and Venous Thrombus in Nonmetastatic Clear-cell Renal Cell Carcinoma: Analysis of the REMEMBER Registry and Implications for Adjuvant Therapy.

BACKGROUND: Further stratification of the risk of recurrence of clear-cell renal cell carcinoma (ccRCC) with venous tumor thrombus (VTT) will facilitate selection of candidates for adjuvant therapy. OBJECTIVE: To assess the impact of tumor grade discrepancy (GD) between the primary tumor (PT) and VTT in nonmetastatic ccRCC on disease-free survival (DFS), overall survival (OS), and cancer-specific survival (CSS). DESIGN, SETTING, AND PARTICIPANTS: This was a retrospective analysis of a multi-institutional nationwide data set for patients with pT3N0M0 ccRCC who underwent radical nephrectomy and thrombectomy. OUTCOMES MEASUREMENTS AND STATISTICAL ANALYSIS: Pathology slides were centrally reviewed. GD, a bidirectional variable (upgrading or downgrading), was numerically defined as the VTT grade minus the PT grade. Multivariable models were built to predict DFS, OS, and CSS. RESULTS AND LIMITATIONS: We analyzed data for 604 patients with median follow-up of 42 mo (excluding events). Tumor GD between VTT and PT was observed for 47% (285/604) of the patients and was an independent risk factor with incremental value in predicting the outcomes of interest (all p < 0.05). Incorporation of tumor GD significantly improved the performance of the ECOG-ACRIN 2805 (ASSURE) model. A GD-based model (PT grade, GD, pT stage, PT sarcomatoid features, fat invasion, and VTT consistency) had a c index of 0.72 for DFS. The hazard ratios were 8.0 for GD = +2 (p < 0.001), 1.9 for GD = +1 (p < 0.001), 0.57 for GD = -1 (p = 0.001), and 0.22 for GD = -2 (p = 0.003) versus GD = 0 as the reference. According to model-converted risk scores, DFS, OS, and CSS significantly differed between subgroups with low, intermediate, and high risk (all p < 0.001). CONCLUSIONS: Routine reporting of VTT upgrading or downgrading in relation to the PT and use of our GD-based nomograms can facilitate more informed treatment decisions by tailoring strategies to an individual patient's risk of progression. PATIENT SUMMARY: We developed a tool to improve patient counseling and guide decision-making on other therapies in addition to surgery for patients with the clear-cell type of kidney cancer and tumor invasion of a vein.

A nomogram based on the preoperative neutrophil-to-lymphocyte ratio to distinguish sarcomatoid renal cell carcinoma from clear cell renal cell carcinoma.

Objective: Our study aimed to assess the predictive value of the preoperative neutrophil-to-lymphocyte ratio(NLR) in distinguishing sarcomatoid renal cell carcinoma (SRCC) from clear cell renal cell carcinoma(CCRCC) and to developing a nomogram based on the preoperative NLR and other factors to distinguish SRCC from CCRCC. Materials and methods: The database involved 280 patients, including 46 SRCC and 234 CCRCC. logistic analysis was conducted to select the variables associated with identifying SRCC preoperatively, and subgroup analysis was used to further validate the ability of NLR with preoperative identification of SRCC.In addition, The data were randomly separated into a training cohort(n=195) and a validation cohort(n=85). And an NLR-based nomogram was plotted based on the logistic analysis results. The nomogram was evaluated according to its discrimination, consistency, and clinical benefits. Results: Multivariate analysis indicated that NLR, flank pain, tumor size, and total cholesterol(TC) were independent risk factors for identifying SRCC. The results of subgroup analysis showed that higher NLR was associated with a higher probability of SRCC in most subgroups. The area under the curve(AUC) of the training and validation cohorts were 0.801 and 0.738, respectively. The results of the calibration curve show high consistency between predicted and actual results. Decision Curve Analysis(DCA) showed clinical intervention based on the model was beneficial over most of the threshold risk range. Conclusion: NLR is a potential indicator for preoperative differentiation of SRCC and CCRCC, and the predictive model constructed based on NLR has a good predictive ability. The new model could provide suggestions for the early identification of SRCC.

Development and validation of a prognostic model incorporating tumor thrombus grading for nonmetastatic clear cell renal cell carcinoma with tumor thrombus: A multicohort study.

There is significant variability with respect to the prognosis of nonmetastatic clear cell renal cell carcinoma (ccRCC) patients with venous tumor thrombus (VTT). By applying multiregion whole-exome sequencing on normal-tumor-thrombus-metastasis quadruples from 33 ccRCC patients, we showed that metastases were mainly seeded by VTT (81.8%) rather than primary tumors (PTs). A total of 706 nonmetastatic ccRCC patients with VTT from three independent cohorts were included in this study. C-index analysis revealed that pathological grading of VTT outperformed other indicators in risk assessment (OS: 0.663 versus 0.501-0.610, 0.667 versus 0.544-0.651, and 0.719 versus 0.511-0.700 for Training, China-Validation, and Poland-Validation cohorts, respectively). We constructed a risk predicting model, TT-GPS score, based on four independent variables: VTT height, VTT grading, perinephric fat invasion, and sarcomatoid differentiation in PT. The TT-GPS score displayed better discriminatory ability (OS, c-index: 0.706-0.840, AUC: 0.788-0.874; DFS, c-index: 0.691-0.717, AUC: 0.771-0.789) than previously reported models in risk assessment. In conclusion, we identified for the first-time pathological grading of VTT as an unheeded prognostic factor. By incorporating VTT grading, the TT-GPS score is a promising prognostic tool in predicting the survival of nonmetastatic ccRCC patients with VTT.

Sarcomatoid-associated gene risk index for clear cell renal cell carcinoma.

Sarcomatoid renal cell carcinoma is a de-differentiated form of kidney cancer with an extremely poor prognosis. Genes associated with sarcomatoid differentiation may be closely related to the prognosis of renal cell carcinoma. The prognosis of renal cell carcinoma itself is extremely variable, and a new prognostic model is needed to stratify patients and guide treatment. Data on clear cell renal cell carcinoma with or without sarcomatoid differentiation were obtained from TCGA database, and a sarcomatoid-associated gene risk index (SAGRI) and column line graphs were constructed using sarcomatoid-associated genes. The predictive power of the SAGRI and column line graphs was validated using an internal validation set and an independent validation set (E-MTAB-1980). The SAGRI was constructed using four sarcoma-like differentiation-related genes, COL7A1, LCTL, NPR3, ZFHX4, and had a 1-year AUC value of 0.725 in the training set, 0.712 in the internal validation set, and 0.770 in the independent validation set for TCGA training cohort, with high model reliability. The molecular characteristics among the SAGRI subgroups were analyzed by multiple methods, and results suggested that the SAGRI-HIGH subgroup may benefit more from immunotherapy to improve prognosis. SAGRI satisfactorily predicted the prognosis of patients with clear cell renal cell carcinoma with or without sarcomatoid differentiation.

Development and validation of prognostic nomograms and a web-based survival rate calculator for sarcomatoid renal cell carcinoma in pre- and post-treatment patients.

BACKGROUND: To develop a clinical prediction model and web-based survival rate calculator to predict the overall survival (OS) and cancer-specific survival (CSS) of sarcomatoid renal cell carcinoma (SRCC) for clinical diagnosis and treatment. METHODS: SRCC patient data were retrieved from Surveillance, Epidemiology, and End Results (SEER) database. Factors independently associated with survival were identified by a Cox regression analysis. Nomograms of the prediction model were constructed using a SEER training cohort and validated with a SEER validation cohort. At the same time, the decision analysis curve, receiver operating characteristic curve, and calibration curve were also used to examine and evaluate the model. A web-based survival rate calculator was constructed to help assist in the assessment of the disease condition and clinical prognosis. RESULTS: The records of 2,742 SRCC cases were retrieved from SEER, while 1,921 cases with a median OS of 14 and CSS of 32 months were used as the training cohort. The developed nomograms were more accurate than that of the American Joint Committee on Cancer staging (C-indexes of 0.767 versus 0.725 for OS and 0.775 versus 0.715 for CSS), with better discrimination than that of the American Joint Committee on Cancer (AJCC) stage model and the calibration was validated in the SEER validation cohort. The model's 3- and 5-year OS and CSS were superior to AJCC and T staging on the analysis decision curve. The prognosis prediction of SRCC established by the prediction model could be evaluated through the web-based survival rate calculator, which plays a guiding role in clinical treatment. CONCLUSIONS: Nomograms and a web-based survival rate calculator predicting the OS and CSS of SRCC patients with better discrimination and calibration were developed.

Circulating Th22 cells, as well as Th17 cells, are elevated in patients with renal cell carcinoma.

T-helper (Th) 22 cells serve an essential role in different types of tumors and autoimmune diseases. No research has been conducted to study the role of Th22 cells in the pathogenesis of renal cell carcinoma (RCC). We aimed to evaluate the prognostic value of circulating Th22, Th17, and Th1 cells in RCC patients. Thirty-two newly diagnosed RCC patients and thirty healthy controls were enlisted in the research. Their peripheral blood was collected, and the frequencies of circulating Th22, Th17, and Th1 cells were detected by flow cytometry. Plasma IL-22 concentrations were examined by an enzyme-linked immunosorbent assay (ELISA). Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to identify the mRNA expression levels of aromatic hydrocarbon receptor (AHR) and RAR-associated orphan receptor C (RORC) in peripheral blood mononuclear cells (PBMC). Compared with the healthy control group, the frequency of circulating Th22 and Th17 cells and concentrations of plasma IL-22 were significantly increased in RCC patients. However, there was no significant difference in the frequency of Th1 cells. A positive correlation between Th22 cells and plasma IL-22 levels was found in RCC patients. Also, there was a significant positive correlation between Th22 and Th17 cells in RCC patients. An up-regulated expression of AHR and RORC transcription factors were also observed in RCC patients. As tumor stage and grade progressed, the frequencies of Th22 and Th17 cells and the level of plasma IL-22 significantly increased. Meanwhile, there was a positive correlation between Th22 and Th17 cells and RCC tumor stage or grade. Furthermore, patients with high Th22 or Th17 cells frequency displayed a decreased trend in survival rate. Our research indicated that the increased circulating Th22 and Th17 cells and plasma IL-22 may be involved in the pathogenesis of RCC and may be involved in the occurrence and development of tumors. Th22 cells, plasma IL-22, and Th17 cells may be promising new clinical biomarkers and may be used as cellular targets for RCC therapeutic intervention.

Liver kinase B1 inhibits smooth muscle calcification via high mobility group box 1.

Vascular calcification is a common pathological feature of atherosclerosis, chronic kidney disease, vascular injury, and aging. Liver kinase B1 (LKB1) plays pivotal roles in cellular processes such as apoptosis, metabolism, and cell cycle regulation. In addition, growing evidence has indicated that LKB1 functions as a tumor suppressor gene. However, its role in vascular calcification has not been reported. LKB1flox/flox mice were hybridized with SM22-CreERT2 transgenic mice and adult mice received tamoxifen to obtain smooth muscle-specific LKB1-knockout (LKB1SMKO) mice. LKB1 expression was decreased under calcifying conditions, and LKB1 overexpression had a protective effect on vascular calcification. However, high mobility group box 1 (HMGB1) overexpression partially counteracted the promotion of vascular calcification induced by LKB1 overexpression. Mechanically, LKB1 could bind to HMGB1 to promote HMGB1 degradation. Furthermore, LKB1SMKO mice showed intensified vascular calcification, which was alleviated by treatment with the HMGB1 inhibitor glycyrrhizic acid. Based on our results, LKB1 may inhibit vascular calcification via inhibiting HMGB1 expression.

Metastatic renal cell carcinoma patients of T4 stage who are in status of N1 stage or older than 76 years cannot benefit from cytoreductive nephrectomy.

BACKGROUND: We aimed to identify which part of the patients with metastatic renal cell carcinoma (mRCC) is not suitable for cytoreductive nephrectomy (CN). METHODS: The data of mRCC patients was acquired from the Surveillance, Epidemiology, and End Results (SEER) database. Multivariate cox regression analysis and nomogram were performed for selecting factors independently associated with survival. Propensity score matching (PSM) was applied to reduce potential bias when comparing survival of mRCC patients treated by CN or non-surgery (NS). The survival analysis of subgroups was estimated by the Kaplan-Meier method and compared by log-rank testing. The summary of subgroup analysis was showed by forest plots. RESULTS: The records of 21,411 patients with mRCC were obtained from the SEER database. After screening, a total of 6532 patients were included for further analysis, of which 6043 underwent CN and 489 underwent NS. Age, T stage, N stage and tumor size were involved in subgroup analysis by PSM according to the result of multivariate cox regression analysis and clinical experience. Survival benefit was not found in T4 stage patients. Further analysis showed that T4&N1 and T4&age ≥ 76 yr subgroups could not obtain survival benefit from CN. CONCLUSION: CN should not be performed in T4 stage mRCC patients who were in status of N1 stage or older than 76 years, because surgery cannot take significant survival benefit for them.

A Population Study to Identify Candidates for Cytoreductive Nephrectomy in Patients with Metastatic Sarcomatoid Renal Cell Carcinoma from the Surveillance, Epidemiology, and End Results (SEER) Database.

BACKGROUND This population study aimed to identify suitable candidates for cytoreductive nephrectomy in patients with metastatic sarcomatoid renal cell carcinoma (RCC) from the US Surveillance, Epidemiology, and End Results (SEER) database. MATERIAL AND METHODS Demographic and clinical data from 1,229 patients with metastatic sarcomatoid RCC were retrieved from the SEER database. Patients were divided into the cytoreductive nephrectomy group (n=937) and the no surgery group (n=292). Multivariate Cox regression analysis identified factors associated with overall survival (OS) and propensity score matching identified factors that significantly impacted the OS. Survival of propensity score-matched subgroups of patients with metastatic sarcomatoid RCC treated by cytoreductive nephrectomy or no surgery was determined by the Kaplan-Meier method and compared by the log-rank test. RESULTS Of the 1,229 patients with metastatic sarcomatoid RCC retrieved from the SEER database, age, tumor size, T stage, and N stage were independent risk factors for patient survival. There were no significant differences in age, N stage, and tumor size between the cytoreductive nephrectomy-treated and non-surgically treated T stage cases following propensity score matching. OS benefits were found in cases with stage T1 (12 months increase), T2 (7.5 months increase), T3a (2 months increase), and T4 (3 months increase), but not in the T3b or T3c subgroups treated by cytoreductive nephrectomy, compared with patients with no surgical treatment. CONCLUSIONS Data from the SEER database showed that cytoreductive nephrectomy improved OS in patients with T1 and T2 metastatic sarcomatoid RCC with a significant long-term survival benefit of >6 months.

Propensity-score matched comparison of partial versus radical nephrectomy for T1N0M0 sarcomatoid renal cell carcinoma.

BACKGROUND: Sarcomatoid renal cell carcinoma is a rare variant of renal cell carcinoma associated with poor clinical outcomes. Currently no reliable evidence indicates whether partial nephrectomy can be an effective treatment for patients with T1N0M0 sarcomatoid renal cell carcinoma. The present study was conducted to compare the overall and cancer-specific survival with partial and radical nephrectomy for such cases. METHODS: Data of sarcomatoid renal cell carcinoma were retrieved from Surveillance, Epidemiology, and End Results database. Factors independently associated with the overall or cancer-specific survival were identified by Cox regression analysis. The overall and cancer-specific survival of propensity-score matched T1N0M0 sarcomatoid renal cell carcinoma patients treated by partial and radical nephrectomy were estimated by the Kaplan-Meier method. RESULTS: The records of 452 T1N0M0 sarcomatoid renal cell carcinoma cases were retrieved. One hundred and fifty-five of these patients underwent partial nephrectomy, while the remaining 297 underwent radical nephrectomy. The median follow-up period in partial-treated group was 38 months (ranging from 2 to 132 months), while that in RN-treated was 39 months (ranging from 0 to 150 months). Age, sex, marital status and tumor size were independent risk factors for the overall and cancer-specific survival. There were no significant differences among age, sex, marital status and tumor size in the PN- and RN-treated T1N0M0 cases according to propensity-score matching. The estimated median overall survival of partial-treated group was 132 months, while that for RN-treated cases was 100 months (P=0.11). The median cancer-specific survival was not reached in both groups (P=0.092). CONCLUSIONS: The overall and cancer-specific survival of T1N0M0 patients treated by partial nephrectomy was not inferior to that of patients treated by radical nephrectomy.

Mathematical modelling of the role of Endo180 network in the development of metastatic bone disease in prostate cancer.

Metastatic bone disease (MBD) is a common complication of advanced cancer and recent research suggests that Endo180 expression is dysregulated through the TGFß-TGFßR-SMAD2/3 signalling pathway during the invasion of tumour cells in the development of MBD. We here provide a model for the dysregulation of the Endo180 network to demonstrate its vital contribution to bone destruction as well as tumour cell growth. The model consisted of a set of ordinary differential equations and reconstructed variations in the bone cells, resultant bone volume, and biochemical factors involved in the TGFß-TGFßR-SMAD2/3 signalling pathway over time. The model also investigated the underlying mechanism in which the change of TGFß affects the TGFß-TGFßR-SMAD2/3 signalling pathway and the resultant Endo180 expression in osteoblastic and tumour cells. The model links the appearance of tumour cells with the inhibition of TGFß binding to its receptors on osteoblastic cells, to affect TGFß-TGFßR-SMAD2/3 signalling and Endo180 expression. Temporal variation in bone cells, bone volume, and the biochemical factors involved in the TGFß-TGFßR-SMAD2/3 pathway as demonstrated in the model simulations agree with published experimental data. The model can be refined based on further discoveries but allows the influence of Endo180 network dysregulation on bone remodelling in MBD to be established. This model could aid in the development of Endo180 targeted therapies for MBD in the future.

Identification of microenvironment-related genes with prognostic value in clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney tumor. Previous studies have shown that the interaction between tumor cells and microenvironment has an important impact on prognosis. Immune and stromal cells are two vital components of the tumor microenvironment. Our study aimed to better understand and explore the genes involved in immune/stromal cells on prognosis. We used the Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data algorithm to calculate immune/stromal scores. According to the scores, we divided ccRCC patients from The Cancer Genome Atlas database into low and high groups and identified the genes which were differentially expressed and significantly associated with prognosis. The result of functional enrichment analysis and protein-protein interaction networks indicated that these genes mainly were involved in extracellular matrix and regulation of cellular activities. Then another independent cohort from the International Cancer Genome Consortium database was used to validate these genes. Finally, we acquired a list of microenvironment-related genes that can predict prognosis for ccRCC patients.

Downregulation of HDGF inhibits the tumorigenesis of bladder cancer cells by inactivating the PI3K-AKT signaling pathway.

BACKGROUND: Hepatoma-derived growth factor (HDGF) is a heparin-binding protein that has been observed to be abnormally expressed in numerous malignancies, but the definite role of HDGF in bladder cancer (BCa) has not been clarified. Here, we conduct the present study to evaluate correlations between HDGF and BCa. METHODS: Bioinformatics analysis was used to evaluate HDGF expression levels in BCa tissues. The effect of HDGF on cell proliferation, migration, invasion, cell cycle and apoptosis was analyzed utilizing CCK-8, clone formation, Transwell assays and flow cytometry, respectively. In addition, the xenograft tumor model was established. RESULTS: Based on bioinformatics analysis, we noticed that HDGF was highly expressed in BCa tissues and was positively correlated with poor prognosis in patients. Knockdown of HDGF markedly reduced tumorigenesis in BCa cells. Furthermore, the results of flow cytometry showed that HDGF deletion enhanced apoptosis in T24 and 253J cells and led to cell cycle arrest in G1 phase. In further studies, we found that tumor growth was inhibited in xenograft nude mouse models with HDGF deletion. The results of RNA-seq analysis revealed that the PI3K-AKT signaling pathway-related genes were obviously changed in HDGF-deficient 253J cells, and this result was further confirmed by Western blot analysis. CONCLUSION: In summary, we suggest that HDGF plays a substantial role in BCa and promotes tumor development and progression by regulating the PI3K-AKT signaling pathway, which provides a promising target for BCa treatment.

Elevated Serum Sialic Acid Levels Predict Prostate Cancer As Well As Bone Metastases.

Objective: To evaluate the value of serum sialic acid (SA) in diagnosis of benign prostatic hyperplasia (BPH), prostate cancer (PCa), and bone metastases in PCa patients. Materials and Methods: Data from 540 patients who were newly diagnosed with PCa or BPH between November 2014 and March 2018 were retrospectively collected and analyzed. Pretreatment SA levels were compared across various groups, then, associations between SA levels and clinic parameters of patients were analyzed as well. Univariate and multivariate logistic regression analyses were further used to identify independent associations. Results: The mean SA levels in patients with PCa were significantly higher than with BPH (p = 0.013). Furthermore, PCa patients with bone metastases showed elevated SA levels compared with PCa without bone metastases (p < 0.001). A multivariate logistic regression model showed that: SA level > 52.35 mg/dL was identified to be independently associated with the diagnosis of PCa (HR = 1.645, p = 0.036), and SA level > 59 mg/dL was identified to be independent association with the presence of bone metastases in PCa patients (HR = 6.421, p = 0.012). Conclusions: Elevated SA level is an independent predictor of prostate cancer as well as its bone metastases. Therefore, SA level may be a promising diagnostic and prognostic biomarker for prostate cancer and bone metastases.

Serum Cystatin C Level Is Not a Promising Biomarker for Predicting Clinicopathological Characteristics of Bladder Urothelial Tumors.

The role of cystatin C (Cys-C) in tumorigenesis and progression of bladder urothelial tumors (BUT) is still indefinite. We retrospectively collected the clinical information from the records of 425 BUT patients. Pretreatment serum Cys-C levels were compared across the various groups. Then we subgroup the patients with GFR ≥ 90 mg/min/1.73 m2, to exclude the effects of lower renal function on cystatin C. No statistically significant differences in the levels of serum Cys-C were found among the tumor characteristics (all P > 0.05). In conclusion, circulating Cys-C was not a reliable predictor for clinicopathological characteristics of BUT patients.

Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9.

Hepatoma-derived growth factor (HDGF) is commonly over-expressed and plays critical roles in the development and progression in a variety of cancers. It has previously been shown that HDGF is overregulated in prostate cancer cells compared to normal prostate cells, which is correlated with cellular migration and invasion of prostate cancer. Here, the molecular mechanisms of HDGF in prostate cancer is investigated. It is shown that HDGF knockdown reduces prostate cancer cellular migration and invasion in both androgen-sensitive LNCaP cells and androgen-insensitive DU145 and PC3 cells. Furthermore, Western blot analysis reveals that HDGF knockdown inhibits epithelial-mesenchymal transition (EMT) of prostate cancer cells by upregulation of protein E-cadherin and downregulation of proteins N-cadherin, Vimentin, Snail and Slug. In addition, mechanistic studies reveal that proteins MMP2 and MMP9 are down-regulated. In conclusion, our data suggested that HDGF knockdown inhibits cellular migration and invasion in vitro of prostate cancer via modulating epithelial-mesenchymal transition (EMT) signaling pathway, as well as MMP2 and MMP9 signaling pathway. These results supported that HDGF is a relevant protein in the progression of prostate cancer and may serve as a potentially therapeutic target for prostate cancer as well as its downstream targets.

Combined thermosensitive in situ gel with AMD3100 in sutureless technique improves the survival and function of kidney transplants in mice.

The mouse is an optimal animal model for kidney transplantation. Recent reports suggest that application of poloxamer 407, a thermosensitive in situ gel, during the sutureless technique significantly increases animal survival, compared to traditional methods. However, further improvement of this technology is greatly needed but remains unexplored. Here, we detected significant inflammation at the region of ureter anastomosis, after kidney transplantation using poloxamer 407. Since chemokines play a pivotal role during inflammation, we implanted an Alzet osmotic pump that gradually releases AMD3100 (a specific inhibitor of the binding of stromal cell-derived factor 1 (SDF-1) to its receptor, CXCR4) at the site of ureter anastomosis in mice that had undergone kidney transplantation. We found that AMD3100 significantly reduced local inflammation, significantly improved animal survival after kidney transplantation, and significantly improved kidney function. Together, these data suggest that inhibition of chemokine signaling at the site of ureter anastomosis may substantially improve animal survival after kidney transplantation through suppression of suturing-related inflammation.

Synergistic antitumor responses by combined GITR activation and sunitinib in metastatic renal cell carcinoma.

Sunitinib, a multitargeted tyrosine kinase inhibitor, is the frontline therapy for renal and gastrointestinal cancers. In view of its well-documented proapoptotic and immunoadjuvant properties, we speculate that combination of Sunitinib and immunotherapy would provide a synergistic antitumor effect. Here, we report that a remarkably synergistic antitumor responses elicited by the combined treatment of Sunitinib and an agonistic antibody against glucocorticoid-induced TNFR related protein (GITR) in a model of metastatic renal cell carcinoma. Sunitinib significantly increased the infiltration, activation, and proliferation and/or cytotoxicity of CD8(+) T cells and NK cells in liver metastatic foci when combined with the anti (α)-GITR agonist, which was associated with treatment-induced prominent upregulation of Th1-biased immune genes in the livers from mice receiving combined therapy versus single treatment. Sunitinib/α-GITR treatment also markedly promoted the maturation, activation and cytokine production of liver-resident macrophages and DCs compared with that achieved by α-GITR or Sunitinib treatment alone in mice. Cell depletion experiments demonstrated that CD8(+) T cells, NK cells and macrophage infiltrating liver metastatic foci all contribute to the antitumor effect induced by combined treatment. Furthermore, mechanistic investigation revealed that Sunitinib treatment reprograms tumor-associated macrophages toward classically activated or "M1" polarization upon GITR stimulation and consequently mounts an antitumor CD8(+) T and NK cell response via inhibiting STAT3 activity. Thus, our findings provide a proof of concept that Sunitinib can synergize with α-GITR treatment to remodel the tumor immune microenvironment to trigger regressions of an established metastatic cancer.

miR-96 suppresses renal cell carcinoma invasion via downregulation of Ezrin expression.

BACKGROUND: The present study examined the role of microRNA (miR)-96 in renal cell carcinoma (RCC) invasion. METHODS: The expression of miR-96 was detected by quantitative reverse transcription-polymerase chain reaction in human RCC cell lines with high (Caki-1) and low (786-O) metastatic potential. Invasive ability and Ezrin expression were assessed in Caki-1 and 786-O cells transfected with a miR-96 mimic or inhibitor using wound healing assays, Transwell assays and western blotting. Expression of miR-96 and Ezrin was also examined in primary RCC samples from 17 patients with metastatic disease and 46 patients who maintained remission during a follow-up period of 37 months. RESULTS: miR-96 expression was significantly lower in Caki-1compared to786-O cells. The invasive ability of Caki-1 and 786-O cells increased following transfection of cells with miR-96 inhibitor, whereas it decreased following transfection with miR-96 mimic. Ezrin levels were negatively correlated with miR-96 in RCC, and inhibition of Ezrin expression suppressed the miR-96-induced change in invasive ability. The negative correlation between miR-96 and metastasis/Ezrin expression was also observed in human RCC specimens. CONCLUSIONS: These results suggest that miR-96 suppresses RCC invasion by modulating Ezrin expression.