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Rapid Identification of DUX4::IGH Fusion in Acute Lymphoblastic Leukemia
Kyoko Moritani1, Minenori Eguchi-Ishimae1, Mari Tezuka-Kagajo1
1Department of Pediatrics Ehime University Graduate School of Medicine Toon Japan.
Insights
DUX4 rearrangement in acute lymphoblastic leukemia (ALL) can be detected using multiplex PCR. This method identifies the DUX4::IGH fusion, a key indicator in a specific ALL subgroup.
Area of Science:
- Hematology
- Molecular Biology
- Oncology
Background:
- DUX4 rearrangement is observed in a subset of B-precursor acute lymphoblastic leukemia (ALL), associated with a favorable prognosis.
- Detecting DUX4 rearrangement is challenging due to the repetitive nature of its gene loci and fusion mechanisms, often requiring whole transcriptome analysis.
- Surface markers CD2 and CD371 can serve as hallmarks for DUX4-rearranged ALL.
Purpose of the Study:
- To establish a more accessible method for identifying DUX4 rearrangement in ALL.
- To investigate the utility of multiplex PCR in detecting the common DUX4::IGH fusion.
- To correlate the presence of DUX4::IGH fusion with specific immunophenotypic markers.
Main Methods:
- Multiplex polymerase chain reactions (PCRs) were designed using primers targeting the DUX4 and IGH gene loci.
- The PCR strategy focused on detecting the DUX4::IGH fusion, which accounts for over 90% of DUX4 fusions.
- Immunophenotyping for CD2 and CD371 expression was performed on ALL cases.
Main Results:
- DUX4::IGH fusion was identified in 3 out of 50 ALL cases using standard PCR.
- Variable insertion of DUX4 into the IGH locus and sequences of unknown origin at the fusion junction were observed, suggesting V(D)J recombination involvement.
- All DUX4::IGH positive cases expressed CD2 and CD371, reinforcing their role as reliable markers.
Conclusions:
- A simple multiplex PCR strategy can effectively identify DUX4::IGH fusion in ALL.
- The findings support the use of CD2 and CD371 as reliable surrogate markers for DUX4::IGH fusion.
- This simplified detection method aids in the diagnosis and characterization of this ALL subgroup.
Introduction:
DUX4 is rearranged and overexpressed in a subgroup of acute lymphoblastic leukemia (ALL) with B-precursor phenotype, with a favorable outcome. Even though characteristic gene expression signature as well as surface expression of CD2/CD371 could be a hallmark of DUX4-rearranged ALL, actual detection of DUX4 rearrangement is, however, largely dependent on whole transcriptome analysis due to the highly repetitive nature of DUX4 gene loci and insertion as the main mechanism of fusion gene formation.
Methods:
Polymerase chain reactions (PCRs) with several combinations of multiplex primers located on DUX4 and IGH gene loci were used for the detection of DUX4::IGH, which represents more than 90% of DUX4 fusion.
Results:
DUX4::IGH fusion was successfully detected in three out of 50 ALL cases analyzed by standard PCR, and these positive cases showed variable insertion of DUX4 into the IGH locus. In all patients, sequences of unknown origin were observed at the junction of DUX4 and IGH sequences, indicating the role of the V(D)J recombination mechanism in fusion gene formation. Although DUX4 is tandemly repeated at its locus, only a single copy of the DUX4 sequence was detected at the IGH locus in two of the three DUX4-rearranged ALL cases. As previously reported, both CD2 and CD371 were positive in all cases with DUX4::IGH fusion, suggesting that the combination of CD2 and CD371 could be a more reliable marker for detecting the presence of this fusion.
Conclusions:
Identification of DUX4::IGH fusion in ALL could be possible more easily by a simple multiplex PCR strategy.
