Multiplex fusion gene testing in pediatric acute myeloid leukemia

Pediatr Int. 2018 Jan;60(1):47-51. doi: 10.1111/ped.13451.

Abstract

Background: Gene abnormalities, particularly chromosome rearrangements generating gene fusion, are associated with clinical characteristics and prognosis in pediatric acute myeloid leukemia (AML). Karyotyping is generally performed to enable risk stratification, but the results are not always consistent with those of reverse transcription-polymerase chain reaction (RT-PCR), and more accurate and rapid methods are required.

Methods: A total of 487 samples from de novo AML patients enrolled in the Japanese Pediatric Leukemia/Lymphoma Study Group (JPLSG) AML-05 study (n = 448), and from acute promyelocytic leukemia (APL) patients enrolled in the JPLSG AML-P05 study (n = 39) were available for this investigation. Multiplex quantitative RT-PCR was performed to detect eight important fusion genes: AML1(RUNX1)-ETO(RUNX1T1), CBFB-MYH11, MLL(KMT2A)-AF9(MLLT3), MLL-ELL, MLL-AF6(MLLT4), FUS(TLS)-ERG, NUP98-HOXA9, and PML-RARA.

Results: Fusion genes were detected in 207 (46.2%) of the 448 AML-05 patient samples. After exclusion of two samples with PML-RARA, no chromosomal abnormalities were identified on karyotyping in 19 of 205 patients (9.3%) positive for fusion genes on RT-PCR. Fusion genes were confirmed on fluorescence in situ hybridization (FISH) in 11 of these 19 patients. In contrast, fusion genes were detected in 37 of 39 patients (94.9%) from the AML-P05 study, and 33 of these results were consistent with the karyotyping. There were discrepancies in four patients (10.8%), three with normal karyotypes and one in whom karyotyping was not possible. All four of these patients were PML-RARA positive on FISH.

Conclusions: Multiplex quantitative RT-PCR-based fusion gene screening may be effective for diagnosis of pediatric AML.

Keywords: Japanese Pediatric Leukemia/Lymphoma Study Group; acute myeloid leukemia; diagnosis; fusion gene; multiplex polymerase chain reaction.

MeSH terms

  • Adolescent
  • Biomarkers, Tumor / genetics*
  • Child
  • Child, Preschool
  • Female
  • Genetic Testing / methods*
  • Humans
  • Infant
  • Karyotyping
  • Leukemia, Myeloid, Acute / diagnosis*
  • Leukemia, Myeloid, Acute / genetics
  • Male
  • Multiplex Polymerase Chain Reaction*
  • Oncogene Fusion*
  • Oncogene Proteins, Fusion / genetics*
  • Prognosis
  • Retrospective Studies
  • Reverse Transcriptase Polymerase Chain Reaction*

Substances

  • Biomarkers, Tumor
  • Oncogene Proteins, Fusion