The transforming proteins of acute promyelocytic leukaemias (APL) are fusions of the promyelocytic leukaemia (PML) and the promyelocytic leukaemia zinc-finger (PLZF) proteins with retinoic acid receptor-alpha (RARalpha). These proteins retain the RARalpha DNA- and retinoic acid (RA)-binding domains, and their ability to block haematopoietic differentiation depends on the RARalpha DNA-binding domain. Thus RA-target genes are downstream effectors. However, treatment with RA induces differentiation of leukaemic blast cells and disease remission in PML-RARalpha APLs, whereas PLZF-RARa APLs are resistant to RA. Transcriptional regulation by RARs involves modifications of chromatin by histone deacetylases, which are recruited to RA-target genes by nuclear co-repressors. Here we show that both PML-RARalpha and PLZF-RARalpha fusion proteins recruit the nuclear co-repressor (N-CoR)-histone deacetylase complex through the RARalpha CoR box. PLZF-RARalpha contains a second, RA-resistant binding site in the PLZF amino-terminal region. High doses of RA release histone deacetylase activity from PML-RARalpha, but not from PLZF-RARalpha. Mutation of the N-CoR binding site abolishes the ability of PML-RARalpha to block differentiation, whereas inhibition of histone deacetylase activity switches the transcriptional and biological effects of PLZF-RARalpha from being an inhibitor to an activator of the RA signalling pathway. Therefore, recruitment of histone deacetylase is crucial to the transforming potential of APL fusion proteins, and the different effects of RA on the stability of the PML-RARalpha and PLZF-RARalpha co-repressor complexes determines the differential response of APLs to RA.
Dasatinib and nilotinib are tyrosine kinase inhibitors (TKIs) developed to overcome imatinib resistance in Philadelphiapositive leukemias. To assess how BcrAbl kinase domain mutation status evolves during sequential therapy with these TKIs and which mutations may further develop and impair their efficacy, we monitored the mutation status of 95 imatinib-resistant patients before and during treatment with dasatinib and/or nilotinib as second or third TKI. We found that 83% of cases of relapse after an initial response are associated with emergence of newly acquired mutations. However, the spectra of mutants conferring resistance to dasatinib or nilotinib are small and nonoverlapping, except for
IntroductionResistance to the tyrosine kinase inhibitor (TKI) imatinib mesylate (IM) in chronic myeloid leukemia (CML) and Philadelphiapositive (Ph ϩ ) acute lymphoblastic leukemia (ALL) is often caused by selection of mutations in the Bcr-Abl kinase domain (KD), [1][2][3][4][5][6][7][8][9] altering residues that are directly or indirectly critical for IM binding. To overcome this problem, novel TKIs have been developed. Dasatinib and nilotinib [10][11][12][13] have been the first ones to enter clinical evaluation and to receive marketing approval in IMresistant patients. Preclinical experience with these agents showed that they are active against several IM-resistant Bcr-Abl mutants, with the exception of T315I. 10,11,14 However, both TKIs have been hypothesized to retain their own "Achilles heels." Recent studies have tried to profile mutations that will probably emerge under dasatinib and nilotinib, inducing random mutagenesis and then selecting for cell clones retaining viability when cultured in the presence of the inhibitors. [15][16][17][18] Results suggested that, besides T315I, other mutants might be critical. To assess how Bcr-Abl KD mutation status evolves under the selective pressure of sequential therapy with novel TKIs and which mutations among those predicted by in vitro studies may indeed develop in patients who relapse on dasatinib or nilotinib, we have monitored the mutation status of 95 IM-resistant patients before and during sequential treatment with one or both of these agents.
Methods
Patients and definitionsThis report focuses on 95 patients (Table 1) who were referred to our laboratory for mutation analysis at the time of IM failure and who received up to 2 subsequent TKIs (dasatinib and/or nilotinib). Thirty-eight patients had chronic-phase (CP) CML, 46 patients had advanced-phase CML (accelerated-phase [AP], n ϭ 11; myeloid blast crisis [BC], n ϭ 18; lymphoid BC, n ϭ 17), and 11 patients had Ph ϩ ALL. For CML patients, IM failure was defined according to European LeukemiaNet recommendations. 19 Similar criteria were applied to define IM failure in Ph ϩ ALL. All 95 patients received a second TKI (dasatinib, n ϭ 55; nilotinib, n ϭ 40). Twenty-six of 95 patients received a third TKI after relapse on second TKI (dasatinib, n ϭ 16; nilotinib, n ϭ 10). For the purpose of this analysis, response to dasat...
An adult patient affected by b 0 -thalassemia major underwent allogeneic bone marrow transplant (BMT) from a matched related donor. Forty days after transplant, allogeneic engraftment failure and autologous b 0 -thalassemic bone marrow recovery were documented. Red blood cell transfusions were required until 118 days post-transplant. Thereafter, the haemoglobin (Hb) levels stabilized over 11.8 gr/dl throughout the ongoing 34-month follow-up, abolishing the need for transfusion support. The Hb electrophoresis showed 100% Hb Fetal (HbF). This unexplained case suggests full HbF production may occur in an adult patient with b 0 -thalassemia major.Homozygous b-thalassemia is a severe disorder caused by inheritance of two b-thalassemia alleles. The gene mutations produce absent or insufficient synthesis of b-globin chains leading to excess a-globin chain accumulation and precipitation in early erythroid cells [1]. Concurrent genetic factors (hereditary persistence of fetal hemoglobin(HbF), a-gene mutations, and db-thalassemia determinants) are known to modify the globin chain imbalance [1,2].To date, the only cure for thalassemia major is stem cell transplantation [3]. Here, we report an atypical case of sustained and full HbF production after graft failure in an adult b 0 -thalassemic patient.On September 2005, an 18-year-old patient affected by b 0 -thalassemia major was referred to our center for bone marrow transplant (BMT) from an HLA identical sibling. The thalassemia Major diagnosis was evident by 2 years of age when the blood transfusion therapy was started and regularly administered every 2-3 weeks. After splenectomy, performed when the patient was 6 years old, the transfusion requirement was reduced to one red blood cell unit every 2-3 months. Such behavior was maintained during the following years without any transfusion-free period. The iron-chelation therapy was never administered.At the time of presentation to our Institution, the patient presented typical thalassemic facies and hepatomegaly (over 4 cm below the ribs). The complete blood counts showed: low Hb level (9.7 g/dl), low red blood cell count (3.59 3 10 12 /l), a mean corpuscular volume of 82 fl, a mild increase of reticulocyte count (4.07%), increased white cell count (3.901 3 10 12 /l), abundant circulating nucleated red cells (80% of total nucleated cells), and an elevated platelet count (803 3 10 9 /l). The bone marrow exhibited marked hypercellularity. The Hb profile showed predominantly HbF (73.6%), and the genetic analysis detected a double mutation in the b-globin locus: the homozygous mutation IVS-I-1 of the b-globin gene and the homozygous mutation for T variant at position 2158 upstream of the Gg-globin gene. High-performance liquid chromatography (HPLC) analyses detected the lack of b-chain synthesis (b 0%) and 2.47% of the a/non a imbalance.Chronic hepatitis C was documented by a HCV-RNA test. Liver biopsy showed mild fibrosis (1/6 degree) and marked iron overload ([3/4] histological score). The liver iron concentration was marked ...
To evaluate the normalization of lymphocyte subsets several years after autologous peripheral blood stem cell transplantation (aPBSCT) and to detect any differences based on the underlying lymphoproliferative diseases, we analyzed the immunological recovery of 149 patients with Non Hodgkin's Lymphoma (NHL), Hodgkin's Disease (HD), Multiple Myeloma (MM). Lymphocyte recovery was assessed before the transplant, on days 15, 30, 60, 90, 120 and on years 1, 2, 4, 6. Analysis of a total of 709 lymphocytes, including total lymphocyte count, CD3 +, CD4 +, CD8 +, CD4 +/CD8 + ratio, CD19 +, CD3 + HLA-DR +, CD16 + 56 +, was performed. The normalization of total lymphocyte counts was achieved between days 14 to 22 following PBSCT. CD3 + cells count showed a normalization after 2 years in the HD and NHL groups and after 4 years in MM group. CD4 + subset achieved normalization during the sixth year in the 3 groups. The CD8 + and CD19 + lymphocytes subsets achieved normal values in the 3 groups at day 60 and at day 120 respectively. CD16 + 56 + and CD3 +/HLA-DR + lymphocytes showed median values above the normal range starting from day 30. Immunological recovery was similar in all 3 groups. Moreover, the recovery of all subsets evaluated was similarly demonstrated within 6 years after aPBSCT.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.