Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma

Susan A J Vaziri, Emmanuel J. Tavares, Ali R. Golshayan, Brian I. Rini, Hakan Aydin, Ming Zhou, Linda Sercia, Laura Wood, Mahrukh K. Ganapathi, Ronald M. Bukowski, Ram Ganapathi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In sporadic clear cell renal cell carcinoma (CCRCC), the von Hippel Lindau (VHL) gene is inactivated by mutation or methylation in the majority of primary (P) tumors. Due to differing effects of wild-type (WT) and mutant (MT) VHL gene on downstream signaling pathways regulating angiogenesis, VHL gene status could impact clinical outcome. In CCRCC, comparative genomic hybridization analysis studies have reported genetic differences between paired P and metastatic (M) tumors. We thus sequenced the VHL gene in paired tumor specimens from 10 patients to determine a possible clonal relationship between the P tumor and M lesion(s) in patients with CCRCC. Using paraffin-embedded specimens, genomic DNA from microdissected samples (>80% tumor) of paired P tumor and M lesions from all 10 patients, as well as in normal tissue from 6 of these cases, was analyzed. The DNA was used for PCR-based amplification of each of the 3 exons of the VHL gene. Sequences derived from amplified samples were compared to the wild-type VHL gene sequence (GenBank Accession No. AF010238). Methylation status of the VHL gene was determined using VHL methylation-specific PCR primers after DNA bisulfite modification. In 4/10 (40%) patients the VHL gene status differed between the P tumor and the M lesion. As expected, when the VHL gene was mutated in both the P tumor and M lesion, the mutation was identical. Further, while the VHL genotype differed between the primary tumor in different kidneys or multiple metastatic lesions in the same patient, the VHL germline genotype in the normal adjacent tissue was always wild-type irrespective of the VHL gene status in the P tumor. These results demonstrate for the first time that the VHL gene status can be different between paired primary and metastatic tissue in patients with CCRCC.

Original languageEnglish (US)
Article numberArticle 51
JournalFrontiers in Oncology
Volume2 MAY
DOIs
StatePublished - Dec 1 2012

Fingerprint

Renal Cell Carcinoma
Genotype
Genes
Neoplasms
Methylation
Polymerase Chain Reaction
Mutation
DNA Primers
Comparative Genomic Hybridization
DNA
Nucleic Acid Databases
Paraffin
Exons
Kidney

Keywords

  • Genetic heterogeneity
  • Renal cancer
  • VHL genotype

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma. / Vaziri, Susan A J; Tavares, Emmanuel J.; Golshayan, Ali R.; Rini, Brian I.; Aydin, Hakan; Zhou, Ming; Sercia, Linda; Wood, Laura; Ganapathi, Mahrukh K.; Bukowski, Ronald M.; Ganapathi, Ram.

In: Frontiers in Oncology, Vol. 2 MAY, Article 51, 01.12.2012.

Research output: Contribution to journalArticle

Vaziri, SAJ, Tavares, EJ, Golshayan, AR, Rini, BI, Aydin, H, Zhou, M, Sercia, L, Wood, L, Ganapathi, MK, Bukowski, RM & Ganapathi, R 2012, 'Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma', Frontiers in Oncology, vol. 2 MAY, Article 51. https://doi.org/10.3389/fonc.2012.00051
Vaziri, Susan A J ; Tavares, Emmanuel J. ; Golshayan, Ali R. ; Rini, Brian I. ; Aydin, Hakan ; Zhou, Ming ; Sercia, Linda ; Wood, Laura ; Ganapathi, Mahrukh K. ; Bukowski, Ronald M. ; Ganapathi, Ram. / Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma. In: Frontiers in Oncology. 2012 ; Vol. 2 MAY.
@article{cedfc11a6b834a779ee6eb5aea30723b,
title = "Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma",
abstract = "In sporadic clear cell renal cell carcinoma (CCRCC), the von Hippel Lindau (VHL) gene is inactivated by mutation or methylation in the majority of primary (P) tumors. Due to differing effects of wild-type (WT) and mutant (MT) VHL gene on downstream signaling pathways regulating angiogenesis, VHL gene status could impact clinical outcome. In CCRCC, comparative genomic hybridization analysis studies have reported genetic differences between paired P and metastatic (M) tumors. We thus sequenced the VHL gene in paired tumor specimens from 10 patients to determine a possible clonal relationship between the P tumor and M lesion(s) in patients with CCRCC. Using paraffin-embedded specimens, genomic DNA from microdissected samples (>80{\%} tumor) of paired P tumor and M lesions from all 10 patients, as well as in normal tissue from 6 of these cases, was analyzed. The DNA was used for PCR-based amplification of each of the 3 exons of the VHL gene. Sequences derived from amplified samples were compared to the wild-type VHL gene sequence (GenBank Accession No. AF010238). Methylation status of the VHL gene was determined using VHL methylation-specific PCR primers after DNA bisulfite modification. In 4/10 (40{\%}) patients the VHL gene status differed between the P tumor and the M lesion. As expected, when the VHL gene was mutated in both the P tumor and M lesion, the mutation was identical. Further, while the VHL genotype differed between the primary tumor in different kidneys or multiple metastatic lesions in the same patient, the VHL germline genotype in the normal adjacent tissue was always wild-type irrespective of the VHL gene status in the P tumor. These results demonstrate for the first time that the VHL gene status can be different between paired primary and metastatic tissue in patients with CCRCC.",
keywords = "Genetic heterogeneity, Renal cancer, VHL genotype",
author = "Vaziri, {Susan A J} and Tavares, {Emmanuel J.} and Golshayan, {Ali R.} and Rini, {Brian I.} and Hakan Aydin and Ming Zhou and Linda Sercia and Laura Wood and Ganapathi, {Mahrukh K.} and Bukowski, {Ronald M.} and Ram Ganapathi",
year = "2012",
month = "12",
day = "1",
doi = "10.3389/fonc.2012.00051",
language = "English (US)",
volume = "2 MAY",
journal = "Frontiers in Oncology",
issn = "2234-943X",
publisher = "Frontiers Media S. A.",

}

TY - JOUR

T1 - Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma

AU - Vaziri, Susan A J

AU - Tavares, Emmanuel J.

AU - Golshayan, Ali R.

AU - Rini, Brian I.

AU - Aydin, Hakan

AU - Zhou, Ming

AU - Sercia, Linda

AU - Wood, Laura

AU - Ganapathi, Mahrukh K.

AU - Bukowski, Ronald M.

AU - Ganapathi, Ram

PY - 2012/12/1

Y1 - 2012/12/1

N2 - In sporadic clear cell renal cell carcinoma (CCRCC), the von Hippel Lindau (VHL) gene is inactivated by mutation or methylation in the majority of primary (P) tumors. Due to differing effects of wild-type (WT) and mutant (MT) VHL gene on downstream signaling pathways regulating angiogenesis, VHL gene status could impact clinical outcome. In CCRCC, comparative genomic hybridization analysis studies have reported genetic differences between paired P and metastatic (M) tumors. We thus sequenced the VHL gene in paired tumor specimens from 10 patients to determine a possible clonal relationship between the P tumor and M lesion(s) in patients with CCRCC. Using paraffin-embedded specimens, genomic DNA from microdissected samples (>80% tumor) of paired P tumor and M lesions from all 10 patients, as well as in normal tissue from 6 of these cases, was analyzed. The DNA was used for PCR-based amplification of each of the 3 exons of the VHL gene. Sequences derived from amplified samples were compared to the wild-type VHL gene sequence (GenBank Accession No. AF010238). Methylation status of the VHL gene was determined using VHL methylation-specific PCR primers after DNA bisulfite modification. In 4/10 (40%) patients the VHL gene status differed between the P tumor and the M lesion. As expected, when the VHL gene was mutated in both the P tumor and M lesion, the mutation was identical. Further, while the VHL genotype differed between the primary tumor in different kidneys or multiple metastatic lesions in the same patient, the VHL germline genotype in the normal adjacent tissue was always wild-type irrespective of the VHL gene status in the P tumor. These results demonstrate for the first time that the VHL gene status can be different between paired primary and metastatic tissue in patients with CCRCC.

AB - In sporadic clear cell renal cell carcinoma (CCRCC), the von Hippel Lindau (VHL) gene is inactivated by mutation or methylation in the majority of primary (P) tumors. Due to differing effects of wild-type (WT) and mutant (MT) VHL gene on downstream signaling pathways regulating angiogenesis, VHL gene status could impact clinical outcome. In CCRCC, comparative genomic hybridization analysis studies have reported genetic differences between paired P and metastatic (M) tumors. We thus sequenced the VHL gene in paired tumor specimens from 10 patients to determine a possible clonal relationship between the P tumor and M lesion(s) in patients with CCRCC. Using paraffin-embedded specimens, genomic DNA from microdissected samples (>80% tumor) of paired P tumor and M lesions from all 10 patients, as well as in normal tissue from 6 of these cases, was analyzed. The DNA was used for PCR-based amplification of each of the 3 exons of the VHL gene. Sequences derived from amplified samples were compared to the wild-type VHL gene sequence (GenBank Accession No. AF010238). Methylation status of the VHL gene was determined using VHL methylation-specific PCR primers after DNA bisulfite modification. In 4/10 (40%) patients the VHL gene status differed between the P tumor and the M lesion. As expected, when the VHL gene was mutated in both the P tumor and M lesion, the mutation was identical. Further, while the VHL genotype differed between the primary tumor in different kidneys or multiple metastatic lesions in the same patient, the VHL germline genotype in the normal adjacent tissue was always wild-type irrespective of the VHL gene status in the P tumor. These results demonstrate for the first time that the VHL gene status can be different between paired primary and metastatic tissue in patients with CCRCC.

KW - Genetic heterogeneity

KW - Renal cancer

KW - VHL genotype

UR - http://www.scopus.com/inward/record.url?scp=84872620354&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84872620354&partnerID=8YFLogxK

U2 - 10.3389/fonc.2012.00051

DO - 10.3389/fonc.2012.00051

M3 - Article

VL - 2 MAY

JO - Frontiers in Oncology

JF - Frontiers in Oncology

SN - 2234-943X

M1 - Article 51

ER -