Improved Identification of von Hippel-Lindau Gene Alterations in Clear Cell Renal Tumors

Purpose: To provide a comprehensive, thorough analysis of somatic mutation and promoter hypermethylation of the von Hippel-Lindau (VHL) gene in the cancer genome, unique to clear cell renal cancer (ccRCC). Identify relationships between the prevalence of VHL gene alterations and alteration subtypes with patient and tumor characteristics. Experimental Design: As part of a large kidney cancer case-control study conducted in Central Europe, we analyzed VHL mutations and promoter methylation in 205 well-characterized, histologically confirmed patient tumor biopsies using a combination of sensitive, high-throughput methods (endonuclease scanning and Sanger sequencing) and analysis of 11 CpG sites in the VHL promoter. Results: We identified mutations in 82.4% of cases, the highest VHL gene mutation prevalence reported to date. Analysis of 11 VHL promoter CpG sites revealed that 8.3% of tumors were hypermethylated and all were mutation negative. In total, 91% of ccRCCs exhibited alteration of the gene through genetic or epigenetic mechanisms. Analysis of patient and tumor characteristics revealed that certain mutation subtypes were significantly associated with Fuhrman nuclear grade, metastasis, node positivity, and self-reported family history of RCC. Conclusion: Detection of VHL gene alterations using these accurate, sensitive, and practical methods provides evidence that the vast majority of histologically confirmed ccRCC tumors possess genetic or epigenetic alteration of the VHL gene and support the hypothesis that VHL alteration is an early event in ccRCC carcinogenesis. These findings also indicate that VHL molecular subtypes can provide a sensitive marker of tumor heterogeneity among histologically similar ccRCC cases for etiologic, prognostic, and translational studies.

[1]  Y. Nakamura,et al.  Genetic alterations during colorectal-tumor development. , 1988, The New England journal of medicine.

[2]  A. Belldegrun,et al.  Interleukin 2 expanded tumor-infiltrating lymphocytes in human renal cell cancer: isolation, characterization, and antitumor activity. , 1988, Cancer research.

[3]  B. Zbar,et al.  Improved detection of allele loss in renal cell carcinomas after removal of leukocytes by immunologic selection. , 1989, Journal of the National Cancer Institute.

[4]  J. Gnarra,et al.  Identification of the von Hippel-Lindau disease tumor suppressor gene. , 1993, Science.

[5]  S. Shuman,et al.  Novel approach to molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase. , 1994, The Journal of biological chemistry.

[6]  J. Brooks,et al.  Mutations of the VHL tumour suppressor gene in renal carcinoma , 1994, Nature Genetics.

[7]  Germline and somatic mutations in the tyrosine kinase domain of the MET proto-oncogene in papillary renal carcinomas , 1997, Nature Genetics.

[8]  H. Ito,et al.  Mutational state of von Hippel-Lindau and adenomatous polyposis coli genes in renal tumors. , 1997, Oncology.

[9]  W. Linehan,et al.  Improved detection of germline mutations in the von Hippel‐Lindau disease tumor suppressor gene , 1998, Human mutation.

[10]  P. Schirmacher,et al.  VHL alterations in human clear cell renal cell carcinoma: association with advanced tumor stage and a novel hot spot mutation. , 2000, Cancer research.

[11]  C. Miller,et al.  Purification, cloning, and characterization of the CEL I nuclease. , 2000, Biochemistry.

[12]  Is the P25L a "real" VHL mutation? , 2001 .

[13]  B. Zbar,et al.  Random mutagenesis‐PCR to introduce alterations into defined DNA sequences for validation of SNP and mutation detection methods , 2001, Human mutation.

[14]  Maria Merino,et al.  Mutations in a novel gene lead to kidney tumors, lung wall defects, and benign tumors of the hair follicle in patients with the Birt-Hogg-Dubé syndrome. , 2002, Cancer cell.

[15]  M. Esumi,et al.  Biallelic inactivation of the von Hippel-Lindau tumor suppressor gene in sporadic renal cell carcinoma. , 2002, The Journal of urology.

[16]  W. Chow,et al.  Global increases in kidney cancer incidence, 1973–1992 , 2002, European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation.

[17]  Y. Nagashima,et al.  Somatic von Hippel–Lindau disease gene mutation in clear‐cell renal carcinomas associated with end‐stage renal disease/acquired cystic disease of the kidney , 2002, Genes, chromosomes & cancer.

[18]  A. Paetau,et al.  Germline mutations in FH predispose to dominantly inherited uterine fibroids, skin leiomyomata and papillary renal cell cancer , 2002, Nature Genetics.

[19]  Holger Moch,et al.  VHL mutations and their correlation with tumour cell proliferation, microvessel density, and patient prognosis in clear cell renal cell carcinoma , 2002, The Journal of pathology.

[20]  W Marston Linehan,et al.  Genetic Basis of Cancer of the Kidney , 2006 .

[21]  P. Choyke,et al.  Mutations in the fumarate hydratase gene cause hereditary leiomyomatosis and renal cell cancer in families in North America. , 2003, American journal of human genetics.

[22]  J. Durocher,et al.  Mutation detection using Surveyor nuclease. , 2004, BioTechniques.

[23]  W. Marston Linehan,et al.  Genetic Basis of Cancer of the Kidney , 2004, Clinical Cancer Research.

[24]  W. Kaelin,et al.  Role of VHL gene mutation in human cancer. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[25]  P. A. van den Brandt,et al.  Prevalence of von Hippel-Lindau gene mutations in sporadic renal cell carcinoma: results from the Netherlands cohort study , 2005, BMC Cancer.

[26]  Tom R. Gaunt,et al.  Genetic Variance Detection: technologies for pharmacogenomics , 2006 .

[27]  W Marston Linehan,et al.  The genetic basis of cancer of the kidney. , 2003, The Journal of urology.

[28]  S. Toyooka,et al.  Detection of EGFR Gene Mutation in Lung Cancer by Mutant-Enriched Polymerase Chain Reaction Assay , 2006, Clinical Cancer Research.

[29]  K. Chew,et al.  Clinical response to therapy targeted at vascular endothelial growth factor in metastatic renal cell carcinoma: impact of patient characteristics and Von Hippel‐Lindau gene status , 2006, BJU international.

[30]  C. Taylor,et al.  Genetic and epigenetic analysis of von Hippel-Lindau (VHL) gene alterations and relationship with clinical variables in sporadic renal cancer. , 2006, Cancer research.

[31]  Thomas E. Hutson.  Targeted Therapy for Renal Cell Carcinoma: A New Treatment Paradigm , 2007, Proceedings.

[32]  R. Figlin,et al.  Sunitinib versus interferon alfa in metastatic renal-cell carcinoma. , 2007, The New England journal of medicine.

[33]  Apurva A Desai,et al.  Sorafenib in advanced clear-cell renal-cell carcinoma. , 2007, The New England journal of medicine.