Nucleophosmin (NPM1) mutations in acute myeloid leukemia (AML) affect exon 12, but sporadically also exon 9 and 11, all causing changes at protein C-terminal end (loss of tryptophans and creation of a nuclear export signal-NES motif) that lead to aberrant cytoplasmic NPM1 (NPM1c+), detectable by immunohistochemistry. Combining immunohistochemistry and molecular analyses in 929 AML patients, we found non-exon 12 NPM1 mutations in 5/387 (1.3%) NPM1c+ cases. Besides mutations in exon 9 (n=1) and exon 11 (n=1), novel mutations in exon 5 were discovered (n=3). One more exon 5 mutation was identified in additional 141 AML patients selected for wild-type NPM1 exon 12. Furthermore, 3 NPM1 rearrangements (i.e. NPM1/RPP30, NPM1/SETBP1, NPM1/CCDC28A) were detected and characterized among 13,979 AML samples screened by cytogenetic/FISH and RNA sequencing. Functional studies demonstrated that in AML cases the new NPM1 proteins harboured an efficient extra NES, either newly created or already present in the fusion partner, ensuring its cytoplasmic accumulation. Our findings support NPM1 cytoplasmic relocation as critical for leukemogenesis and reinforce the role of immunohistochemistry in predicting any AML-associated NPM1 genetic lesions. Also, this study highlights the need for developing new specific assays for molecular diagnosis and monitoring of NPM1-mutated AML.
NOVEL NPM1 EXON 5 MUTATIONS AND GENE FUSIONS LEADING TO ABERRANT CYTOPLASMIC NUCLEOPHOSMIN IN AML / Martelli, Maria Paola; Rossi, Roberta; Venanzi, Alessandra; Meggendorfer, Manja; Perriello, Vincenzo Maria; Martino, Giovanni; Spinelli, Orietta; Ciurnelli, Raffaella; Varasano, Emanuela; Brunetti, Lorenzo; Ascani, Stefano; Quadalti, Corinne; Cardinali, Valeria; Mezzasoma, Federica; Gionfriddo, Ilaria; Milano, Francesca; Pacini, Roberta; Tabarrini, Alessia; Bigerna, Barbara; Albano, Francesco; Specchia, Giorgina; Vetro, Calogero; Di Raimondo, Francesco; Annibali, Ombretta; Avvisati, Giuseppe; Rambaldi, Alessandro; Falzetti, Franca; Tiacci, Enrico; Sportoletti, Paolo; Haferlach, Torsten; Haferlach, Claudia; Falini, Brunangelo. - In: BLOOD. - ISSN 0006-4971. - (2021). [10.1182/blood.2021012732]