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We previously reported that histone deacetylase (HDAC) activity is elevated, but is not correlated to the JAK-2 mutation status, in patients with myelofibrosis myeloid metaplasia (MMM) (Blood 107:319b 2005). Now we have studied more patients: totally, 17 with MMM, 19 with other myeloproliferative disorders (MPD), and 16 normal volunteers as controls. Significantly elevated HDAC levels again was shown in patients with MMM compared with other MPD patients and normal volunteer controls (p

ErbB2, c-Myc and p53 gene expressions are widely reported for the poor prognostic markers for medulloblastoma (MB) patients. Recently, Wnt pathway and Sonic Hedgehog (SHH) pathways are known to related to the oncogenesis of MB cells, and the PDGF receptor gene and some adhesion molecule gene expressions are also known to the markers of the dissemination or the metastasis of MB cells. For adult malignant brain tumors, the molecular targeted therapy of the antibody (trastuzumab, cetuximab), the tyrosine kinase inhibitor (imatinib, gefinitib) or other signal transduction inhibitor (cyclopamin) are clinically researched. We examined several important marker gene expressions of the molecular therapy in various children’s brain tumors and searched the possible molecular targeted therapy. [Materials and methods] Fifteen children’s brain tumor (five MB, seven glioma, three ependymoma) and four adult brain tumor (two glioma, two glioblastoma) were examind. The three MB cases were primary metaststic ones. The mRNA expressions of the marker genes (ErbB2, PDGF receptor, PCNA, SPARC, β-catenin, SUFU, c-Myc, p53, TrkC and so on) were examined by quantity polymerase chain (qPCR) reaction with fresh frozen tumor cells. For the normal control, we used the normal cerebellum total RNA samples of the Becton, Dikinson and Company. CYBR green coloring system was used for the qPCR and their primers were designed in the region between two exons. GAPDH gene expression was also examined as an internal control and all of the gene expression were normalized by GAPDH expression. [Results] ErbB2 gene expression in MB cells were various, but their expression were similar to clinical futures, such as, the higher expressed cases had some high risk factors. In glioma cells, ErbB2 gene expression were almost equal by lower levels. PDGF receptor gene expression were more than five to ten times elevated in all of the glioma cells, even if they were low grade malignancy glioma, and were higher than that in MB cells. PCNA was specially elevated in primary metastatic MB cells. [Conclusion] Our data shows that the molecular characteristics of the children’s brain tumors were thought to be different by cases, even if they had same histopathological characters. Some special gene tageting therapy, such as anti-ErbB2, PDGFR and/or PCNA therapy can be used for the various children’s brain tumors which are resistant for conventional therapies and are cured by adequate molecular therapy.

Background: The mutational status of immunoglobulin VH genes (IgVH) is an important prognostic marker in chronic lymphocytic leukemia (CLL), but its determination remains unadapted to routine practice. Several reports have showed that ZAP-70, whose expression can be detected by flow cytometry, can be considered as a reliable surrogate marker of IgVH mutational status. We recently conducted a gene expression profiling study of 18 cases of CLL, which pointed out 2 other genes which might also discriminate CLL groups: the lipoprotein lipase (LPL) and the disintegrin and metalloprotease 29 (ADAM29) genes which were overexpressed preferentially in unmutated (UM) and mutated (MT) cases respectively. Methods: We analyzed frozen cells obtained at diagnosis for 127 CLL patients (87 Binet stage A, 40 stage B or C). LPL, ADAM29 and house keeping GAPDH gene expression were measured by real time quantitative polymerase chain reaction in 111 cases, and ZAP-70 protein by flow cytometry in 107 cases, both analyses being performed in 93 cases. LPL and ADAM29 were also quantified in peripheral blood mononuclear cells (PBMC, n=4) and purified B cells (n=3) of healthy individuals. We correlated the results with the previously determined IgVH mutational status and clinical outcome. Results: With a cut-off value determined to be 1 for the LPL/GAPDH copy number ratio, LPL expression had a positive predictive value (PPV) of 68% for UM cases and a negative predictive value (NPV) of 85% for MT patients. Alternatively ADAM29 expression (ADAM29/GAPDH > 3) had a PPV of 77% for MT cases and of 86% for UM cases. Combining LPL and ADAM29 RNA quantifications by a simple 1:1 ratio (L/A ratio; threshold=1) provided slightly better results than those obtained with ZAP-70 (positivity threshold = 20%) for PPV of UM status (90% vs 76%) and similar results for NPV of MT status (90% vs 91%). Simultaneaous usage of L/A ratio and ZAP-70 expression allowed an almost perfect (73/74) assessment of the IgVH status in 80% (74/93) of patients with concordant results (L/A+, ZAP-70+ or L/A-, ZAP-70-). Serial measurements of L/A ratio and ZAP-70 expression showed that both parameters can change over time (median follow-up 38 months, range 6–159) in a fraction of patients (5/25 tested). ADAM29 was not detected while LPL was present at very low levels or absent in PMBC or purified B cells from healthy donors. The IgVH mutational status, ZAP-70 and L/A ratio were predictive of event-free survival for the whole cohort and for stage A patients. However only the L/A ratio was significantly associated with a shorter survival (p=0.03) for stage B/C patients. Conclusions: Combination of LPL and ADAM29 mRNA quantification provides a good surrogate marker of the IgVH mutational status in CLL patients. Used in association with ZAP-70, it represents an reliable alternative to the sequencing of IgVH genes. In addition, it might constitute a more powerful prognostic marker than IgVH mutational status or ZAP-70.