Signaling pathway-based stratification of clear cell renal cell carcinoma.

434 Background: There are marked differences in responses to therapy among patients with clear cell renal cell carcinoma (ccRCC), which makes the outcome difficult to predict. This study is aimed to define a new classification system that would elucidate distinctions between carcinomas in order to facilitate selection of the appropriate treatment. We mapped cell signaling pathways in individual renal cell carcinomas and identified different classes based on commonly shared phosphorylation-driven signaling networks. Methods: Laser capture microdissection and reverse-phase protein arrays were used to profile 75 key nodes in 16 primary clear cell renal cancers. These nodes represent many signaling pathways known to be important in tumorigenesis and progression. Results: Statistical analysis revealed significant differences (p <0.05) in signaling levels between two groups of samples, group A (4 samples) and group B (12 samples), for 27 of the 75 endpoints tested. In group A, high activation levels of EGFR, RET, and RASGFR1 converged to activate AKT/mTOR. Group B, showed high phosphorylation levels of ERK1/2 and STAT transcription factors and samples significantly partitioned in two clusters of 7 and 5 cases designated C and D. Group C showed elevated expression of a regulator of autophagy, LC3B; group D showed activation of Src and STAT transcription factors, suggesting the presence of cytokine-mediated cell survival pathways. A DNA copy number analysis was performed on the same samples and the results showed that group B represents some paradigmatic cases of ccRCC, with VHL loss-of-function mutations. Conclusions: The proteins identified appeared to be linked to pathways that are targeted by drugs typically used to treat clear cell renal cell carcinoma. Thus, this type of analysis could be useful for stratifying patients and selecting the best therapeutic approaches.

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Positive feedback regulation of lncRNA PVT1 and HIF2α contributes to clear cell renal cell carcinoma tumorigenesis and metastasis

Oncogene ◽

10.1038/s41388-021-01971-7 ◽

2021 ◽

Author(s):

Ming-xiao Zhang ◽

Li-zhen Zhang ◽

Liang-min Fu ◽

Hao-hua Yao ◽

Lei Tan ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Positive Feedback ◽

Renal Cell ◽

Clear Cell ◽

Biological Significance ◽

Specific Inhibitor ◽

Migration And Invasion ◽

Loss Of Function ◽

Cell Renal Cell Carcinoma

AbstractLong noncoding RNAs (lncRNAs) have been reported to exert important roles in tumors, including clear cell renal cell carcinoma (ccRCC). PVT1 is an important oncogenic lncRNA which has critical effects on onset and development of various cancers, however, the underlying mechanism of PVT1 functioning in ccRCC remains largely unknown. VHL deficiency-induced HIF2α accumulation is one of the major factors for ccRCC. Here, we identified the potential molecular mechanism of PVT1 in promoting ccRCC development by stabilizing HIF2α. PVT1 was significantly upregulated in ccRCC tissues and high PVT1 expression was associated with poor prognosis of ccRCC patients. Both gain-of-function and loss-of function experiments revealed that PVT1 enhanced ccRCC cells proliferation, migration, and invasion and induced tumor angiogenesis in vitro and in vivo. Mechanistically, PVT1 interacted with HIF2α protein and enhanced its stability by protecting it from ubiquitination-dependent degradation, thereby exerting its biological significance. Meanwhile, HIF2α bound to the enhancer of PVT1 to transactivate its expression. Furthermore, HIF2α specific inhibitor could repress PVT1 expression and its oncogenic functions. Therefore, our study demonstrates that the PVT1/ HIF2α positive feedback loop involves in tumorigenesis and progression of ccRCC, which may be exploited for anticancer therapy.

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Transcription Factors BARX1 and DLX4 Contribute to Progression of Clear Cell Renal Cell Carcinoma via Promoting Proliferation and Epithelial–Mesenchymal Transition

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.626328 ◽

2021 ◽

Vol 8 ◽

Author(s):

Guoliang Sun ◽

Yue Ge ◽

Yangjun Zhang ◽

Libin Yan ◽

Xiaoliang Wu ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Transcription Factors ◽

Cell Carcinoma ◽

Renal Cell ◽

Clear Cell ◽

Epithelial Mesenchymal Transition ◽

Gene Set Enrichment Analysis ◽

Differentially Expressed ◽

Cell Renal Cell Carcinoma ◽

Mesenchymal Transition

Dysregulation of transcription factors contributes to the carcinogenesis and progression of cancers. However, their roles in clear cell renal cell carcinoma remain largely unknown. This study aimed to evaluate the clinical significance of TFs and investigate their potential molecular mechanisms in ccRCC. Data were accessed from the cancer genome atlas kidney clear cell carcinoma cohort. Bioinformatics algorithm was used in copy number alterations mutations, and differentially expressed TFs’ analysis. Univariate and multivariate Cox regression analyses were performed to identify clinically significant TFs and construct a six-TF prognostic panel. TFs’ expression was validated in human tissues. Gene set enrichment analysis (GSEA) was utilized to find enriched cancer hallmark pathways. Functional experiments were conducted to verify the cancer-promoting effect of BARX homeobox 1 (BARX1) and distal-less homeobox 4 (DLX4) in ccRCC, and Western blot was performed to explore their downstream pathways. As for results, many CNAs and mutations were identified in transcription factor genes. TFs were differentially expressed in ccRCC. An applicable predictive panel of six-TF genes was constructed to predict the overall survival for ccRCC patients, and its diagnostic efficiency was evaluated by the area under the curve (AUC). BARX1 and DLX4 were associated with poor prognosis, and they could promote the proliferation and migration of ccRCC. In conclusion, the six-TF panel can be used as a prognostic biomarker for ccRCC patients. BARX1 and DLX4 play oncogenic roles in ccRCC via promoting proliferation and epithelial–mesenchymal transition. They have the potential to be novel therapeutic targets for ccRCC.

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Role of Wnt Signaling Pathways in Clear Cell Renal Cell Carcinoma Pathogenesis in Relation to VHL and HIF Status

Clinical Oncology and Research ◽

10.31487/j.cor.2020.03.09 ◽

2020 ◽

pp. 1-7

Author(s):

Raviprakash T. Sitaram ◽

Börje Ljungberg ◽

Göran Roos ◽

Marene Landström ◽

Raviprakash T. Sitaram

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Signaling Pathways ◽

Renal Cell ◽

Clear Cell ◽

Cell Renal Cell Carcinoma ◽

Genetic Changes ◽

Genetic Abnormalities ◽

Tumor Types

Renal cell carcinoma (RCC) encompasses various tumor types characterized by a variety of genetic abnormalities. The genetic changes, like mutations, deletions, and epigenetic alterations, can affect the signaling components and signaling networks, causing the modification of tumor pathogenesis and prognosis of RCC. The most prevalent RCC, clear cell RCC (ccRCC), is asymptomatic in the early stages, refractory to chemotherapy and radiation therapy, and has a poorer prognosis compared with the papillary and chromophobe ccRCC types. Loss of the VHL gene and upregulation of oxygen sensors, hypoxiainducible factor alphas (HIF-α), which promote different growth factors, is a signature of sporadic ccRCC. The VHL-HIF-α and Wnt/β-catenin pathways are closely connected and contribute to the ontogeny of ccRCC. This review confines to ccRCC and the role of the Wnt/β-catenin signaling pathways and its crosstalk with VHL/HIF.

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VHL substrate transcription factor ZHX2 as an oncogenic driver in clear cell renal cell carcinoma

Science ◽

10.1126/science.aap8411 ◽

2018 ◽

Vol 361 (6399) ◽

pp. 290-295 ◽

Cited By ~ 32

Author(s):

Jing Zhang ◽

Tao Wu ◽

Jeremy Simon ◽

Mamoru Takada ◽

Ryoichi Saito ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Renal Cell ◽

Clear Cell ◽

Nuclear Factor Κb ◽

Loss Of Function ◽

Cell Renal Cell Carcinoma ◽

A Genome

Inactivation of the von Hippel-Lindau (VHL) E3 ubiquitin ligase protein is a hallmark of clear cell renal cell carcinoma (ccRCC). Identifying how pathways affected by VHL loss contribute to ccRCC remains challenging. We used a genome-wide in vitro expression strategy to identify proteins that bind VHL when hydroxylated. Zinc fingers and homeoboxes 2 (ZHX2) was found as a VHL target, and its hydroxylation allowed VHL to regulate its protein stability. Tumor cells from ccRCC patients with VHL loss-of-function mutations usually had increased abundance and nuclear localization of ZHX2. Functionally, depletion of ZHX2 inhibited VHL-deficient ccRCC cell growth in vitro and in vivo. Mechanistically, integrated chromatin immunoprecipitation sequencing and microarray analysis showed that ZHX2 promoted nuclear factor κB activation. These studies reveal ZHX2 as a potential therapeutic target for ccRCC.

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Mir-144-3p Promotes Cell Proliferation, Metastasis, Sunitinib Resistance in Clear Cell Renal Cell Carcinoma by Downregulating ARID1A

Cellular Physiology and Biochemistry ◽

10.1159/000484395 ◽

2017 ◽

Vol 43 (6) ◽

pp. 2420-2433 ◽

Cited By ~ 35

Author(s):

Wen Xiao ◽

Ning Lou ◽

Hailong Ruan ◽

Lin Bao ◽

Zhiyong Xiong ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Proliferation ◽

Cell Carcinoma ◽

Renal Cell ◽

Clear Cell ◽

Xenograft Model ◽

Luciferase Reporter ◽

Loss Of Function ◽

Cell Renal Cell Carcinoma

Background/Aims: We previously performed microRNA (miRNA) microarray to identify effective indicators of clear cell renal cell carcinoma (ccRCC) tissue samples and preoperative/postoperative plasma in which we identified miR-144-3p as an oncomiRNA. However, the molecular mechanism of miR-144-3p remains unclear. This study aims to explore the roles of miR-144-3p in the invasion, migration and Sunitinib-resistance in ccRCC and to elucidate the underlying mechanisms. Methods: Gain and loss of function approaches were used to investigate the cell proliferation, cycle distribution, clonogenicity, migration, invasion, chemosensitivity of miR-144-3p in vitro. The xenograft model was used to assess the effects of miR-144-3p overexpression on tumorigenesis. Bioinformatics analysis and dual-luciferase reporter assay were used to indentify AT-rich interactive domain 1A (ARID1A) as a direct target gene of miR-144-3p. Quantitative RT-PCR, Western blotting, and immunohistochemical (IHC) staining were used to explore ARID1A expression level of the mRNA and protein. Results: We found that miR-144-3p overexpression enhanced cell proliferation, clonogenicity, migration, invasion, and chemoresistance in ccRCC cells. Notably, the oncotumor activities of miR-144-3p were mediated by repressing the expression of ARID1A. The downregulation of ARIDIA could promote the function of miR-144-3p in cell proliferation, metastasis and chemoresistance. Consistently, ARID1A mRNA and protein levels were decreased in ccRCC and in nude mice, and they negatively correlated with miR-144-3p. Conclusion: Higher miR-144-3p may enhance malignancy and resistance to Sunitinib in ccRCC by targeting ARID1A, the observations may uncover novel strategies of ccRCC treatment.

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Spectrum of VHL mutations in clear cell renal cell carcinoma

Advances in molecular oncology ◽

10.17650/2313-805x-2020-7-3-48-57 ◽

2020 ◽

Vol 7 (3) ◽

pp. 48-57

Author(s):

N. N. Mazurenko ◽

I. V. Tsyganova ◽

V. V. Strelnikov ◽

A. V. Balbutsky ◽

T. F. Malivanova ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Renal Cell ◽

Clear Cell ◽

Silent Mutation ◽

Missense Mutations ◽

Loss Of Function ◽

Cell Renal Cell Carcinoma ◽

Vhl Protein ◽

Next Generation Sequencing Ngs

The VHL gene alterations are the early and characteristic feature of clear cell renal cell carcinoma (ccRCC). We have examined VHL mutations in sporadic 98 ccRCC cases to evaluate their localization in relation to functionally important motifs of the VHL protein. The DNA samples were obtained from snap-frozen carcinoma biopsies and used for Sanger sequencing, while 62 ccRCC DNA cases were studied by next generation sequencing (NGS) analysis in parallel. In 73 (74.4 %) оf 98 ccRCC cases the somatic non-silent VHL mutations were identified. Loss of function VHL mutations (nonsilent, frameshifts or in splicing sites) were detected in 40 (40.8 %) ccRCC, while missense mutations – in 35 (35.7 %) ccRCC. In total 76 mutations important for VHL functioning were detected in 72 (73 %) ccRCC samples, of them 15 mutations (deletion / insertion in-frame or frameshifts) were identified for the first time. Four ccRCC cases contained two mutations each. Most of missense mutations disturb the sites of VHL interactions with HIF, РКС or kinesin. The pathogenicity of p.P154P silent mutation and intronic mutations near mRNA VHL splicing sites was discussed. The obtained results are important for understanding the role of VHL mutations in ccRCC progression and prognosis.

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MicroRNAs in clear cell renal cell carcinoma: biological functions and applications

Journal of Kidney Cancer and VHL ◽

10.15586/jkcvhl.2015.40 ◽

2015 ◽

Vol 2 (4) ◽

pp. 140-152 ◽

Cited By ~ 9

Author(s):

Gianluca Aguiari

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Renal Cell ◽

Clear Cell ◽

Causative Role ◽

Loss Of Function ◽

Cell Renal Cell Carcinoma ◽

Small Noncoding Rnas ◽

Potential Applications

MicroRNAs (miRs) are small noncoding RNAs that govern many biological processes. They frequently acquire a gain or a loss of function in cancer and hence play a causative role in the development and progression of neoplasms. They could be used as biomarkers to improve our knowledge on diagnosis, prognosis and drug resistance, and to attempt therapeutic approaches in several types of cancer including clear cell renal cell carcinoma (ccRCC). ccRCC is the most predominant subtype of RCC that accounts for about 90% of all renal cancers. Since ccRCC is generally asymptomatic until very late, it is difficult to diagnose early. Moreover, in the absence of preventive treatments for metastatic ccRCC after surgical resection of the primary cancer, predictive prognostic biomarkers are needed in order to achieve appropriate therapies. Herein the role of miRs in the biology of ccRCC and the potential applications of these molecules are discussed. Moreover, future applications in the diagnostic and prognostic field, as well as their impact on drug response and therapeutic targets are also explored. Their use in clinical practice as molecular biomarkers alone, or in combination with other biological markers, could accelerate progress help design personalized therapies, limit side effects, and improve quality of life of ccRCC patients.

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Comparison of stem cell signaling pathways in long-term responders (LR) versus primary refractory (PR) patients with metastatic clear-cell renal cell carcinoma (ccRCC) under sunitinib (SU) treatment (SULONG study).

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.15_suppl.4578 ◽

2014 ◽

Vol 32 (15_suppl) ◽

pp. 4578-4578

Author(s):

Javier Puente ◽

Nuria Laínez ◽

María José Méndez-Vidal ◽

Emilio Esteban ◽

Daniel E. Castellano ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Stem Cell ◽

Cell Signaling ◽

Cell Carcinoma ◽

Signaling Pathways ◽

Renal Cell ◽

Clear Cell ◽

Cell Renal Cell Carcinoma ◽

Cell Signaling Pathways

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Five-year survival analysis of interferon-a plus sorafenib versus sorafenib monotherapy as first-line treatment for metastatic clear cell renal cell carcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2015.33.7_suppl.486 ◽

2015 ◽

Vol 33 (7_suppl) ◽

pp. 486-486

Author(s):

Ye Ding-Wei ◽

Zhang Hai-Lang

Keyword(s):

Renal Cell Carcinoma ◽

Renal Cell ◽

Clear Cell ◽

Line Treatment ◽

First Line ◽

Cell Renal Cell Carcinoma ◽

Group A ◽

Group B ◽

First Line Treatment

486 Background: We compared the short- and long-term efficacy and safety of interferon-a plus sorafenib with sorafenib monotherapy as first-line treatment in metastatic clear cell renal cell carcinoma (mRCC) patients. Methods: mRCC patients who had not received systemic treatment were administered either interferon-a (300 MIU IM every other day) plus sorafenib (400 mg bid n=24, Group A) or sorafenib monotherapy (400 mg bid; n=24, Group B). Objective responses (OR; RECIST criterion ver 1.0) and differences in OR, progression-free survival (PFS), overall survival (OS) and toxicity were compared. Results: Sixty eight percent males were present in both the groups with comparable baseline demographic characteristics. After a median follow-up of 68 months, the OR was comparable (p=0.726) in Group A vs B; complete remission (1 vs 0 cases), partial remission (6 vs 7 cases), stable disease (14 vs 15 cases), and progressive disease (3 vs 3 cases). No significant difference was observed in the median PFS (p=0.965) and median OS (p=0.223) between both groups [9.4 months (95% CI: 5.8-17.4), Group A vs 14.0 months (95% CI: 9.9-18.0), Group B] and [32.9 months (95% CI: 8.2-87.1), Group A vs 20.4 months (95% CI: 16.2-24.6), Group B], respectively. The 5-year survival rate was higher in Group A vs B (46% vs 25%). Toxicity symptoms like fever (13 vs 3 cases), fatigue (15 vs 9 cases) and neutropenia (6 vs 1 cases) were more pronounced in Group A vs B. The incidence of hand and foot skin reactions, alopecia, rash, hypertension, liver dysfunction, hypophosphatemia, anemia, and other toxicities was similar in both groups. There were 14 (58.3%) and 8 (33.3%) cases of dosage reduction or suspension in Groups A and B, respectively. Conclusions: Short-term effect of interferon-a plus sorafenib as first-line treatment for mRCC was comparable to sorafenib monotherapy in terms of OR and PFS. Although higher toxicity was reported for interferon-a plus sorafenib, the combination holds promise for improving long-term OS.

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