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Downregulated Expression of Genes Mapping on Chromosome 9 in Chronic Myeloid Leukemia Cases Bearing Genomic Deletions on Der(9)

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Downregulated Expression of Genes Mapping on Chromosome 9 in Chronic Myeloid Leukemia Cases Bearing Genomic Deletions on Der(9) Letters to the Editor 813 regimen, with survival of 16 versus 27% respectively, hazard 1Division of Haematology/Oncology, ratio 1.33 (95% CI 1.01–1.77; P ¼ 0.05).8 When interpreted Department of Paediatrics, The Hospital for Sick Children, Toronto, Ontario, Canada; in the context of our meta-analysis results, the trial results 2 questions the role of fludarabine, cytarabine and priming Department of Paediatrics, Program in Child Health Evaluative Sciences, The Hospital for Sick Children, G-CSF in patients with high-risk AML. Toronto, Ontario, Canada; There are several limitations to this meta-analysis. First, the 3Department of Medicine, University Health Network, 95% CI for all survival outcomes other than remission rates does Toronto, Ontario, Canada; not exclude the possibility of clinically meaningful effects. 4Department of Hematology-Oncology, Children’s Mercy However, given the point estimate of a 1% absolute risk Hospitals and Clinics, Kansas City, MO, USA and difference in overall survival, it is unclear whether CSF priming 5Department of Oncology, Children’s Hospital of would warrant prioritization for future study in AML trials, as a Philadelphia, Philadelphia, PA, USA definitive trial would likely require several thousand patients. E-mail: [email protected] Second, the number of studies did not permit meta-regression or further stratified analyses. In addition, there was substantial References heterogeneity in the types of chemotherapy administered concurrent with CSF priming. However, on balance, we did 1 Disis ML. Clinical use of subcutaneous G-CSF or GM-CSF in not find clinically important differences in the effect of CSF malignancy. Oncology (Williston Park) 2005; 19 (4 Suppl 2): 5–9. priming for the variables we examined. 2 Sung L, Nathan PC, Alibhai SM, Tomlinson GA, Beyene J. Meta- In conclusion, the results of this meta-analysis analysis: effect of prophylactic hematopoietic colony-stimulating demonstrate that CSF priming does not improve outcome factors on mortality and outcomes of infection. Ann Intern Med 2007; 147: 400–411. in AML and thus should not be used in routine clinical 3 Becker PS. Growth factor priming in therapy of acute myelogenous care. Although this meta-analysis did not definitively address leukemia. Curr Hematol Rep 2004; 3: 413–418. the question of CSF priming in standard risk patients, the overall 4 Lowenberg B, van Putten W, Theobald M, Gmur J, Verdonck L, results of the analysis do not lend strong support to study this Sonneveld P et al. Effect of priming with granulocyte colony- question further. stimulating factor on the outcome of chemotherapy for acute myeloid leukemia. N Engl J Med 2003; 349: 743–752. 5 Landis JR, Koch GG. The measurement of observer agreement for Acknowledgements categorical data. Biometrics 1977; 33: 159–174. 6 DerSimonian R, Laird N. Meta-analysis in clinical trials. Control Clin Trials 1986; 7: 177–188. LS is supported by a career development award with the Canadian 7 Thomas X, Raffoux E, Botton S, Pautas C, Arnaud P, de Revel T Child Health Clinician Scientist Training Program, a strategic et al. Effect of priming with granulocyte-macrophage colony- program with the Canadian Institutes of Health Research. SMHA is stimulating factor in younger adults with newly diagnosed acute a Research Scientist of the National Cancer Institute of Canada. myeloid leukemia: a trial by the Acute Leukemia French Association JB would like to acknowledge funding from the Canadian Institute (ALFA) Group. Leukemia 2007; 21: 453–461. 8 Milligan DW, Wheatley K, Littlewood T, Craig JI, Burnett AK. of Health Research. Fludarabine and cytosine are less effective than standard ADE chemotherapy in high-risk acute myeloid leukemia, and addition of 1,2 3 2 4 2 L Sung , SMH Alibhai , J Beyene , A Gamis , R Almeida , G-CSF and ATRA are not beneficial: results of the MRC AML-HR S Smith5 and R Aplenc5 randomized trial. Blood 2006; 107: 4614–4622. Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu) Downregulated expression of genes mapping on chromosome 9 in chronic myeloid leukemia cases bearing genomic deletions on der(9) Leukemia (2009) 23, 813–816; doi:10.1038/leu.2008.311; microdeletions could be associated with the loss of tumor published online 6 November 2008 suppressor genes or genes implicated in cell-cycle regulation, resulting in a proliferative advantage of the leukemic clone.4 This phenomenon may act by a gene dosage effect known as Genomic deletions flanking the breakpoint on der(9)t(9;22) haploinsufficiency: mutation or loss of a single allele may be occur in 10–18% of patients with chronic myeloid leukemia sufficient to elicit a cellular phenotype that leads to tumorigen- (CML). These microdeletions have a variable extension and esis without the inactivation of the second allele. involve sequences on chromosomes 9 and/or 22, located The aim of our study was therefore to assess whether the proximally to the ABL and distally to the BCR gene.1 The sequences loss on der(9) in CML patients was associated with a deletions on der(9) are associated with a poor prognosis of gene product decrease. To date, an expression study of genes interferon-a (IFN-a) therapy, whereas controversial data about mapping within the deleted regions on der(9) has never been their influence on the response to imatinib are available.2,3 The performed. We have reported earlier a detailed molecular molecular mechanisms responsible for this unfavorable prog- cytogenetic characterization of 60 (18%) out of 334 CML nosis are still unclear. The most probable hypothesis is that patients, using panels of bacterial artificial chromosome clones Leukemia Letters to the Editor 814 mapping next to the t(9;22) breakpoint.5 We showed that the controls the cellular lifetime together with localization of many extent of the genomic deletion on der(9) ranged from a few critical proteins. The loss of PPP2R4, ASB6, SH3GLB2 and hundred kilobases to many megabases, causing the loss of PKN3 was detected in 60, 57, 43 and 33% of cases with several genes with a known function. deletions, respectively. Noteworthily, the PPP2R4 gene is one of To provide more accurate information about the significance the regulatory subunits of the phosphatase 2A (PP2A) protein, a of the deletion status of these genes with respect to their serine/threonine phosphatase, which acts as a tumor suppressor functional biological meaning, we performed an expression in CML by antagonizing BCR-ABL. The inactivation of PP2A has analysis of 28 genes by quantitative real-time PCR in 30 CML been reported earlier to be associated with disease progression patients (see Supplementary Information for full description of in CML, as its activity in blast crisis patient cells is markedly Materials and methods). Expression data analysis, performed by inhibited.7 ASB6 contains a C-terminal SOCS box motif, typical the relative expression software tool,6 revealed a lower level of of negative regulators of the cytokine receptors. SH3GLB2 is all the 28 examined target gene transcripts in the group of involved in the activation of the apoptotic cell death mechanism deleted patients as compared with CML cases without deletions and in mediating the generation of signaling complexes. PKN3 is (Supplementary Table). However, the difference was statistically one of the target protein kinases for the small GTPase Rho and significant for six genes: protein kinase PKNbeta (PKN3, can be regulated by various signal transduction pathways that P ¼ 0.003), SH3-domain GRB2-like endophilin B2 (SH3GLB2, mediate cell growth and transformation. The subset of genes P ¼ 0.0018), protein phosphatase 2A regulatory subunit B0 mapping on chromosome 22 did not reveal any statistically (PPP2R4, P ¼ 0.007), ankyrin repeat and SOCS box-containing significant downregulation (Supplementary Table). Earlier papers 6 isoform (ASB6, P ¼ 0.002), ubiquitin-specific protease 20 took into consideration the potential role, in the pathogenesis of (USP20, P ¼ 0.010) and torsin family 1 member B (TOR1B, CML with der(9) deletion, of the tumor suppressor gene SWI/SNF- P ¼ 0.009) (Figure 1a, Supplementary Table). The expression related, matrix-associated actin (SMARCB1) gene, which is levels of the downregulated genes were 0.096, 0.222, 0.223, implicated in leukemogenesis through the remodeling of a local 0.198, 0.222 and 0.293 for PKN3, SH3GLB2, PPP2R4, ASB6, chromatin structure.1 In this study, we could not confirm a USP20 and TOR1B, respectively (Supplementary Table). All the significant downregulation of the SMARCB1 gene, excluding its six downregulated genes are located on chromosome 9 involvement in the poor prognosis of this subset of patients. It can sequences, centromeric to the ABL gene (Figure 1b) and are be noted that all six downregulated genes were located inside a implicated in crucial cellular pathways. It can be noted that the region starting at 1 Mb up to 2 Mb centromeric to ABL (Figure 1b). downregulated genes USP20 and TOR1B were lost in 67% of This observation could clarify earlier results showing a worse the 30 CML patients bearing der(9) deletions identified in our outcome for CML patients with large deletions.8 The down- study. Low levels of TOR1B transcripts could confer to the cell a regulation did not affect all these six genes at the same time, but greater vulnerability to oxidative stress, as this protein is at least one of them was significantly downregulated in 19 out of involved in conformational modeling of proteins and protection 21 patients (90%) bearing chromosome 9 sequences loss of from stress. USP20 possesses a deubiquitinating activity and 41 Mb. All the nine patients with a deletion of p1 Mb did not Figure 1 Expression analysis and chromosomal localization of the six downregulated genes. (a) A graphic representation of gene expression data obtained by relative expression software tool (REST) analysis.
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