Despite the recent identification of recurrent SETBP1 mutations in atypical chronic myeloid leukemia (aCML), a complete description of the somatic lesions responsible for the onset of this disorder is still lacking. To find additional somatic abnormalities in aCML, we performed whole-exome sequencing on 15 aCML cases. In 2 cases (13.3%), we identified somatic missense mutations in the ETNK1 gene. Targeted resequencing on 515 hematological clonal disorders revealed the presence of ETNK1 variants in 6 (8.8%) of 68 aCML and 2 (2.6%) of 77 chronic myelomonocytic leukemia samples. These mutations clustered in a small region of the kinase domain, encoding for H243Y and N244S (1/8 H243Y; 7/8 N244S). They were all heterozygous and present in the dominant clone. The intracellular phosphoethanolamine/phosphocholine ratio was, on average, 5.2-fold lower in ETNK1-mutated samples (P < .05). Similar results were obtained using myeloid TF1 cells transduced with ETNK1 wild type, ETNK1-N244S, and ETNK1-H243Y, where the intracellular phosphoethanolamine/phosphocholine ratio was significantly lower in ETNK1-N244S (0.76 ± 0.07) and ETNK1-H243Y (0.37 ± 0.02) than in ETNK1-WT (1.37 ± 0.32; P = .01 and P = .0008, respectively), suggesting that ETNK1 mutations may inhibit the catalytic activity of the enzyme. In summary, our study shows for the first time the evidence of recurrent somatic ETNK1 mutations in the context of myeloproliferative/myelodysplastic disorders. Key Points Whole-exome sequencing reveals the presence of recurrent somatic mutations of ETNK1 in patients with atypical chronic myeloid leukemia. ETNK1 mutations impair the catalytic activity of the enzyme, causing a decrease in the intracellular levels of phosphoethanolamine.

Imatinib currently represents the standard treatment in the early chronic phase of chronic myelogenous leukemia (CML), thanks to the high percentage of cytogenetic complete remission achieved, but it is yet unclear to what extent it can eradicate leukemia. Therefore, different vaccination strategies have been suggested, mainly based on the exploitment of the junctional peptides spanning the fusion region of the Bcr/Abl proteins. To identify new potential immunologic targets, 63 Philadelphia chromosome–positive patients and 6 BCR/ABL–positive cell lines were tested in nested reverse transcriptase PCR to detect the presence of BCR/ABL transcripts arising from the alternative splicing of the main BCR/ABL transcripts. We could detect BCR/ABL transcripts with junctions between BCR exon 1, 13, or 14 and ABL exon 4 in ∼80% of patients and 84% of cell lines, beside the main fusion transcripts. Translation products of these transcripts were characterized at their COOH terminus by a large amino acid portion derived from the out of frame (OOF) reading of ABL gene. These proteins were detected in BCR/ABL–positive cell lines by immunoprecipitation and immunohistochemistry. Finally, we determined whether OOF-specific CD8⁺ T cells could be found in the peripheral blood of CML patients and whether they could acquire effector function following in vitro sensitization with OOF-derived peptides predicted to bind to human leucocyte antigen (HLA)-A2 and HLA-A3 molecules. We detected the presence of OOF-specific CD8⁺ T cells in four of four patients studied, and in one case, these T cells exhibited specific cytotoxic activity against both peptide-pulsed targets and autologous primary CML cells. [Cancer Res 2007;67(11):5300–7]

Chlorhexidine (CHX) is widely used in oral care for its broad-spectrum antimicrobial activity but can cause significant side effects. Sodium DNA has emerged as a potential adjunct capable of modulating cellular responses. This study assessed whether sodium DNA enhances the antibacterial and antibiofilm activity of 0.20% and 0.12% CHX mouthwashes against Streptococcus mutans and Escherichia coli, and evaluated their effects on the viability and phagocytic activity of Dictyostelium discoideum, a model for mammalian phagocytes. All CHX-containing mouthwashes were bactericidal against S.mutans, regardless of sodium DNA, whereas CHX-only formulations were more effective against E.coli in time–kill assays. All formulations inhibited biofilm formation at 50–0.01%. In S. mutans, early biofilms were strongly inhibited (50–0.39%), whereas mature biofilms were less affected. In E. coli, sodium DNA enhanced inhibition of both biofilm formation (50–1.56%) and mature biofilms (50–3.12%). The 0.12% CHX–sodium DNA formulation most effectively modulated D.discoideum viability and phagocytic activity, and metabolomics showed that sodium DNA reduced CHX-induced metabolic stress. This study integrates antimicrobial, antibiofilm, cellular, and metabolomic analyses to assess CHX with sodium DNA. Sodium DNA reduces CHX-induced cytotoxicity and metabolic stress while maintaining antimicrobial activity, offering insights for optimizing oral hygiene formulations through combined microbial and host-cell evaluation.