Detection of the G17V RHOA mutation in angioimmunoblastic T-cell lymphoma and related lymphomas using quantitative allele-specific PCR

PLoS One. 2014 Oct 13;9(10):e109714. doi: 10.1371/journal.pone.0109714. eCollection 2014.

Abstract

Angioimmunoblastic T-cell lymphoma (AITL) and peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) are subtypes of T-cell lymphoma. Due to low tumor cell content and substantial reactive cell infiltration, these lymphomas are sometimes mistaken for other types of lymphomas or even non-neoplastic diseases. In addition, a significant proportion of PTCL-NOS cases reportedly exhibit features of AITL (AITL-like PTCL-NOS). Thus disagreement is common in distinguishing between AITL and PTCL-NOS. Using whole-exome and subsequent targeted sequencing, we recently identified G17V RHOA mutations in 60-70% of AITL and AITL-like PTCL-NOS cases but not in other hematologic cancers, including other T-cell malignancies. Here, we establish a sensitive detection method for the G17V RHOA mutation using a quantitative allele-specific polymerase chain reaction (qAS-PCR) assay. Mutated allele frequencies deduced from this approach were highly correlated with those determined by deep sequencing. This method could serve as a novel diagnostic tool for 60-70% of AITL and AITL-like PTCL-NOS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • DNA Mutational Analysis / methods*
  • Exome
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Lymphoma, T-Cell / classification
  • Lymphoma, T-Cell / diagnosis*
  • Lymphoma, T-Cell / genetics
  • Mutation, Missense*
  • Polymerase Chain Reaction / methods*
  • rhoA GTP-Binding Protein / genetics*

Substances

  • RHOA protein, human
  • rhoA GTP-Binding Protein

Grants and funding

This work was supported by Grants-in-Aid for Scientific Research (KAKENHI) (24390241, 23659482, 23118503, and 22130002 to S.C.; 25461407 to M.S.-Y.), and the Adaptable and Seamless Technology Transfer Program through target-driven R and D (A-STEP) to M.S.-Y. from the Ministry of Education, Culture, Sports, Science and Technology of Japan. This work was also supported by the Mochida Memorial Foundation for Medical and Pharmaceutical Research, and the Uehara Memorial Foundation to M.S.-Y. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.