Next-generation sequencing of translocation renal cell carcinoma reveals novel RNA splicing partners and frequent mutations of chromatin-remodeling genes

Clin Cancer Res. 2014 Aug 1;20(15):4129-40. doi: 10.1158/1078-0432.CCR-13-3036. Epub 2014 Jun 4.

Abstract

Purpose: MITF/TFE translocation renal cell carcinoma (TRCC) is a rare subtype of kidney cancer. Its incidence and the genome-wide characterization of its genetic origin have not been fully elucidated.

Experimental design: We performed RNA and exome sequencing on an exploratory set of TRCC (n = 7), and validated our findings using The Cancer Genome Atlas (TCGA) clear-cell RCC (ccRCC) dataset (n = 460).

Results: Using the TCGA dataset, we identified seven TRCC (1.5%) cases and determined their genomic profile. We discovered three novel partners of MITF/TFE (LUC7L3, KHSRP, and KHDRBS2) that are involved in RNA splicing. TRCC displayed a unique gene expression signature as compared with other RCC types, and showed activation of MITF, the transforming growth factor β1 and the PI3K complex targets. Genes differentially spliced between TRCC and other RCC types were enriched for MITF and ID2 targets. Exome sequencing of TRCC revealed a distinct mutational spectrum as compared with ccRCC, with frequent mutations in chromatin-remodeling genes (six of eight cases, three of which were from the TCGA). In two cases, we identified mutations in INO80D, an ATP-dependent chromatin-remodeling gene, previously shown to control the amplitude of the S phase. Knockdown of INO80D decreased cell proliferation in a novel cell line bearing LUC7L3-TFE3 translocation.

Conclusions: This genome-wide study defines the incidence of TRCC within a ccRCC-directed project and expands the genomic spectrum of TRCC by identifying novel MITF/TFE partners involved in RNA splicing and frequent mutations in chromatin-remodeling genes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATPases Associated with Diverse Cellular Activities
  • Apoptosis
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism
  • Blotting, Western
  • Carcinoma, Renal Cell / genetics*
  • Carcinoma, Renal Cell / metabolism
  • Carcinoma, Renal Cell / pathology
  • Cell Cycle
  • Cell Proliferation
  • Chromatin Assembly and Disassembly / genetics*
  • DNA Helicases / genetics*
  • DNA-Binding Proteins
  • Gene Expression Regulation, Neoplastic
  • Genome-Wide Association Study
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Immunoenzyme Techniques
  • Kidney Neoplasms / genetics
  • Kidney Neoplasms / metabolism
  • Kidney Neoplasms / pathology
  • Mutation / genetics*
  • Mutation Rate
  • Nuclear Proteins
  • RNA Splicing / genetics*
  • RNA, Messenger / genetics
  • RNA-Binding Proteins / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Trans-Activators / genetics
  • Translocation, Genetic / genetics*
  • Tumor Cells, Cultured

Substances

  • Biomarkers, Tumor
  • DNA-Binding Proteins
  • KHDRBS2 protein, human
  • KHSRP protein, human
  • LUC7L3 protein, human
  • Nuclear Proteins
  • RNA, Messenger
  • RNA-Binding Proteins
  • Trans-Activators
  • ATPases Associated with Diverse Cellular Activities
  • DNA Helicases
  • INO80 protein, human

Supplementary concepts

  • Clear-cell metastatic renal cell carcinoma