MLL/GRAF fusion in an infant acute monocytic leukemia (AML M5b) with a cytogenetically cryptic ins(5;11)(q31;q23q23)

Abstract More than 30 fusions involving the MLL gene at 11q23 have been reported in acute myeloid leukemia (AML). Some of these chimeras are rather common, such as MLL/MLLT3 ( AF9 ), but many are quite rare, with some, for example, MLL/GRAF , described only in a single case. The MLL/GRAF fusion, in which the reciprocal hybrid was not expressed, suggesting that the former transcript was the leukemogenic one, was detected in a juvenile myelomonocytic leukemia with a t(5;11)(q31;q23). Here, we report a second case—an infant acute monocytic leukemia (AML M5b)—with an MLL/GRAF fusion. By conventional G‐banding, the karyotype was normal. However, Southern blot and fluorescence in situ hybridization analyses revealed that MLL was rearranged and that the 5′ part of the MLL gene was inserted into 5q in the vicinity of 5q31, which harbors GRAF . Reverse‐transcriptase polymerase chain reaction (PCR) showed that exon 9 of MLL was fused in‐frame with exon 19 of GRAF . Extralong genomic PCR with subsequent sequence analysis demonstrated that the breakpoints occurred in intron 9 of MLL , nine base pairs (bp) downstream from exon 9, and in intron 18 of GRAF , 117 bp downstream from exon 18. A 6‐bp insertion (ACACTC) of unknown origin was present at the junction. The putative MLL/GRAF fusion protein would retain the AT‐hook DNA‐binding domain, the DNA methyl transferase motif, the transcription repression domain of MLL, and the SH3 domain of GRAF. As expected, the reciprocal GRAF/MLL was neither expressed nor generated at the genomic level as a consequence of the ins(5;11)(q31;q23q23). On the basis of the now‐reported two cases with MLL/GRAF , we conclude that this transcript—but not the reciprocal one—characterizes a rare genetic subgroup of infant AML. © 2004 Wiley‐Liss, Inc.