hes1-protein--human and Precursor-T-Cell-Lymphoblastic-Leukemia-Lymphoma

Inhibition of NOTCH1 signaling with gamma-secretase inhibitors (GSIs) has been proposed as a molecularly targeted therapy in T-cell acute lymphoblastic leukemia (T-ALL). However, GSIs seem to have limited antileukemic activity in human T-ALL and are associated with severe gastrointestinal toxicity resulting from inhibition of NOTCH signaling in the gut. Inhibition of NOTCH1 signaling in glucocorticoid-resistant T-ALL restored glucocorticoid sensitivity and co-treatment with glucocorticoids inhibited GSI-induced gut toxicity. Thus, combination therapies with GSIs plus glucocorticoids may offer a new opportunity for the use of anti-NOTCH1 therapies in human T-ALL.

Topics: Adult; Amyloid Precursor Protein Secretases; Animals; Antineoplastic Combined Chemotherapy Protocols; Basic Helix-Loop-Helix Transcription Factors; Child; Drug Resistance, Neoplasm; Drug Synergism; Enzyme Inhibitors; Gastrointestinal Diseases; Gene Expression Regulation, Neoplastic; Glucocorticoids; Homeodomain Proteins; Humans; Intestinal Mucosa; Intestines; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Mice; Mice, Knockout; Neoplasm Proteins; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Protein Processing, Post-Translational; Receptor, Notch1; Transcription Factor HES-1

We have previously shown that elevated expression of Hairy enhancer of split 1 (Hes1) contributes to blast crisis transition in Bcr-Abl-positive chronic myelogenous leukemia. Here we investigate whether Hes1 is involved in the development of other myeloid neoplasms. Notably, Hes1 expression was elevated in only a few cases of 65 samples with different types of myeloid neoplasms. Interestingly, elevated expression of Hes1 was found in two of five samples of Fip1-like1 platelet-derived growth factor receptor-α (FIP1L1-PDGFA)-positive myeloid neoplasms associated with eosinophilia. Whereas FIP1L1-PDGFRα alone induced acute T-cell leukemia or myeloproliferative neoplasms in mouse bone marrow transplantation models, mice transplanted with bone marrow cells expressing both Hes1 and FIP1L1-PDGFRα developed acute leukemia characterized by an expansion of myeloid blasts and leukemic cells without eosinophilic granules. FIP1L1-PDGFRα conferred cytokine-independent growth to Hes1-transduced common myeloid progenitors, interleukin-3-dependent cells. Imatinib inhibited the growth of common myeloid progenitors expressing Hes1 with FIP1L1-PDGFRα, but not with imatinib-resistant FIP1L1-PDGFRα mutants harboring T674I or D842V. In contrast, ponatinib efficiently eradicated leukemic cells expressing Hes1 and the imatinib-resistant FLP1L1-PDGFRΑ mutant in vitro and in vivo. Thus, we have established mouse models of FIP1L1-PDGFRA-positive leukemia in myeloid blast crisis, which will help elucidate the pathogenesis of the disease and develop a new treatment for it.

Topics: Amino Acid Substitution; Animals; Antineoplastic Agents; Basic Helix-Loop-Helix Transcription Factors; Benzamides; Blast Crisis; Female; Gene Expression Regulation, Leukemic; Homeodomain Proteins; Humans; Imatinib Mesylate; Interleukin-3; Leukemia, Myeloid, Acute; Male; Mice; Mice, Inbred BALB C; mRNA Cleavage and Polyadenylation Factors; Mutation, Missense; Neoplasms, Experimental; Oncogene Proteins, Fusion; Piperazines; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Pyrimidines; Receptor, Platelet-Derived Growth Factor alpha; Transcription Factor HES-1

To analyze and investigate the expression levels of HES1, C-MYC and NF-kB in peripheral blood of patients with T cell acute lymphoblastic leukemia (T-ALL) and their significance.. Sixty patients with T-ALL and 60 patients with acute myelogenous leukemia (AML) diagnosed in our hospital from June 2012 to March 2015 were enrolled in T-ALL group and AML group, respectively. Another 30 healthy people were enrolled in the control group. Peripheral blood was collected to detect the expression levels of HES1, C-MYC and NF-kB by RT-PCR. The general data and the expression of HES1, C-MYC and NF-kB in peripheral blood were compared among the patients with different type of leukemia, cytogenetical types and different prognosis.. There was no significant difference in baseline data, such as age and sex among the 3 groups (P＞0.05). The Hb level, WBC and Plt count, BM blast cell ratio in T-ALL and AML groups all were significantly higher than those in control group (P＜0.01), but there were no statistical difference in above-mentioned indicators between T-ALL and AML groups (P＞0.05). The expression levels of HES1, C-MYC and NF-kB in peripheral blood among 3 groups were significantly differenct (P＜0.01), the expressions levels of HES1, C-MYC and NF-kB in T-ALL and AML groups were significantly higher than those in control were significantly group (P＜0.01), moreover, the expression levels of above-mentional indicators in T-ALL groups were significantly higher than than those in AML group (P＜0.01). The expression levels of HES1, C-MYC and NF-kB iin T-ALL patients with poor prognosis were significantly higher than those in T-ALL patients with favorable prognosis (P＜0.01); the expression levels of HES1, C-MYC and NF-kB in peripheral blood of patients with different theraptic efficacy were follow: complete remission group＜partial remission group＜no remission group (P＜0.01).. The HES1, C-MYC and NF-kB are highly expressed in peripheral blood of the patients with T-ALL, moreover, the expression levels maybe different, because of the cytogenetic, and theraptic efficacy.. 急性T淋巴细胞白血病患者外周血HES1、C-MYC和NF-kB表达水平及其意义.. 分析并探讨急性T淋巴细胞白血病（T-ALL）患者外周血HES1，C-MYC和NF-kB表达水平及其意义.. 2012年6月- 2015年3月在本院确诊为T-ALL和急性髓系白血病（AML）患者各60例，分别设为T-ALL组和AML组，另30例健康体检人群作为对照组。采集3组外周血，应用RT-PCR法检测HES1、C-MYC、NF-kB表达水平，对比3组一般资料及不同白血病型、细胞遗传学分型、预后患者外周血HES1、C-MYC、NF-kB的表达差异.. 3组年龄、性别等基线资料比较无差异（P＞0.05）。T-ALL组、AML组血红蛋白（Hb）、白细胞（WBC）、血小板（Plt）、BM细胞（BM blast cell）均显著高于正常对照组（P＜0.01），但T-ALL组与AML组之间差异无统计学意义（P＞0.05）。3组外周血HES1、C-MYC、NF-kB表达水平差异有显著统计学意义（P＜0.01），且在T-ALL组和AML组显著高于对照组（P＜0.01），在T-ALL组中HES1、C-MYC、NF-kB表达水平显著高于AML组（P＜0.01）。T-ALL组中预后不良患者HES1、C-MYC、NF-kB表达水平均显著高于预后良好的T-ALL患者（P＜0.01），完全缓解组HES1、C-MYC、NF-kB表达水平显著高于部分缓解组及未缓解组（P＜0.01）.. HES1、C-MYC、NF-kB在T-ALL患者外周血中呈高表达状态，其表达水平可因细胞遗传学和疗效而有所差异.

Topics: Humans; Leukemia, Myeloid, Acute; NF-kappa B; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Remission Induction; T-Lymphocytes; Transcription Factor HES-1

Notch proteins determine cell fate decisions in the development of diverse tissues. Notch has been initially found in T-ALL but its role has been also studied in myelopoiesis and myeloid leukemias. Studies in different model systems have led to a widespread controversy as to whether Notch promotes or blocks myeloid differentiation. In this work, we evaluated the influence of Notch activation on leukemic cell differentiation along the monocytic and myelocytic pathway induced by phorbol 12-myristate 13-acetate (PMA) or all-trans retinoic acid (ATRA). We observed that differentiation of the human myeloblastic cell line HL-60 can be retarded or blocked by Delta/Notch interaction. ATRA induces complete remission in patients with acute promyelocytic leukemia, but it cannot completely eliminate the leukemic clone and to be effective it should be combined with chemotherapy. Our findings suggest that Notch signaling may contribute to the incomplete elimination of the leukemic cells after PMA or ATRA treatment and the blockage of Notch pathway may be beneficial in the treatment of myeloid leukemia. © 2014 International Society for Advancement of Cytometry.

Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; Cell Differentiation; Cell Line, Tumor; Cell Lineage; Cell Proliferation; Granulocyte Precursor Cells; HL-60 Cells; Homeodomain Proteins; Humans; Jurkat Cells; Leukemia, Promyelocytic, Acute; Mice; Myeloid Cells; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Receptor, Notch1; RNA, Messenger; Signal Transduction; Tetradecanoylphorbol Acetate; Transcription Factor HES-1; Tretinoin; U937 Cells

Oncogenic activation of NOTCH1 signaling plays a central role in the pathogenesis of T-cell acute lymphoblastic leukemia, with mutations on this signaling pathway affecting more than 60% of patients at diagnosis. However, the transcriptional regulatory circuitries driving T-cell transformation downstream of NOTCH1 remain incompletely understood. Here we identify Hairy and Enhancer of Split 1 (HES1), a transcriptional repressor controlled by NOTCH1, as a critical mediator of NOTCH1-induced leukemogenesis strictly required for tumor cell survival. Mechanistically, we demonstrate that HES1 directly downregulates the expression of BBC3, the gene encoding the PUMA BH3-only proapoptotic factor in T-cell acute lymphoblastic leukemia. Finally, we identify perhexiline, a small-molecule inhibitor of mitochondrial carnitine palmitoyltransferase-1, as a HES1-signature antagonist drug with robust antileukemic activity against NOTCH1-induced leukemias in vitro and in vivo.

Topics: Animals; Antineoplastic Agents; Basic Helix-Loop-Helix Transcription Factors; Cell Survival; Cells, Cultured; Gene Expression Regulation, Leukemic; Gene Silencing; Gene Targeting; HEK293 Cells; Homeodomain Proteins; Humans; Jurkat Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Microarray Analysis; Molecular Targeted Therapy; Perhexiline; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Receptor, Notch1; Transcription Factor HES-1

In children, T-cell acute lymphoblastic leukemia (T-ALL) has inferior prognosis compared with B-cell precursor ALL. In order to improve survival, individualized treatment strategies and thus risk stratification algorithms are warranted, ideally already at the time of diagnosis.. We analyzed the frequency and prognostic implication of mutations in NOTCH1 and FBXW7 in 79 cases of Swedish childhood T-ALL treated according to the Nordic Society of Pediatric Hematology and Oncology (NOPHO) ALL-1992 and ALL-2000 protocols. In a subgroup of patients, we also investigated the functional relevance of NOTCH1 mutations measured as expression of the HES1, MYB, and MYC genes.. Forty-seven of the cases (59%) displayed mutations in NOTCH1 and/or FBXW7. There was no difference in overall (P = 0.14) or event-free survival (EFS) (P = 0.10) in patients with T-ALL with mutation(s) in NOTCH1/FBXW7 compared with patients with T-ALL without mutations in any of these genes. T-ALL carrying NOTCH1 mutations had increased HES1 and MYB mRNA expression (HES1 9.2 ± 1.9 (mean ± SEM), MYB 8.7 ± 0.8 (mean ± SEM)) compared to T-ALL with wild-type NOTCH1 (HES1 1.8 ± 0.7, MYB 5.1 ± 1.2, P = 0.02 and 0.008, respectively). In cases of T-ALL with high HES1 expression, improved overall (P = 0.02) and EFS (P = 0.028) was seen.. Increased NOTCH activity, reflected by increased HES1 expression, is associated with improved outcome in pediatric T-ALL, but its role as a diagnostic tool or a therapeutic target in future clinical treatment protocols remains to be elucidated.

Topics: Adolescent; Basic Helix-Loop-Helix Transcription Factors; Cell Cycle Proteins; Child; Child, Preschool; F-Box Proteins; F-Box-WD Repeat-Containing Protein 7; Female; Genes, myb; Homeodomain Proteins; Humans; Infant; Male; Mutation; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Prognosis; Receptor, Notch1; Transcription Factor HES-1; Ubiquitin-Protein Ligases

Topics: Anti-Inflammatory Agents; Basic Helix-Loop-Helix Transcription Factors; Blotting, Western; Chromatin Immunoprecipitation; Dexamethasone; Drug Resistance, Neoplasm; Homeodomain Proteins; Humans; Mutation; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Real-Time Polymerase Chain Reaction; Receptor, Notch1; Receptors, Glucocorticoid; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; Transcription Factor HES-1; Tumor Cells, Cultured

Notch signaling plays both oncogenic and tumor suppressor roles, depending on cell type. In contrast to T-cell acute lymphoblastic leukemia (ALL), where Notch activation promotes leukemogenesis, induction of Notch signaling in B-cell ALL (B-ALL) leads to growth arrest and apoptosis. The Notch target Hairy/Enhancer of Split1 (HES1) is sufficient to reproduce this tumor suppressor phenotype in B-ALL; however, the mechanism is not yet known. We report that HES1 regulates proapoptotic signals by the novel interacting protein Poly ADP-Ribose Polymerase1 (PARP1) in a cell type-specific manner. Interaction of HES1 with PARP1 inhibits HES1 function, induces PARP1 activation, and results in PARP1 cleavage in B-ALL. HES1-induced PARP1 activation leads to self-ADP ribosylation of PARP1, consumption of nicotinamide adenine dinucleotide(+), diminished adenosine triphosphate levels, and translocation of apoptosis-inducing factor from mitochondria to the nucleus, resulting in apoptosis in B-ALL but not T-cell ALL. Importantly, induction of Notch signaling by the Notch agonist peptide Delta/Serrate/Lag-2 can reproduce these events and leads to B-ALL apoptosis. The novel interaction of HES1 and PARP1 in B-ALL modulates the function of the HES1 transcriptional complex and signals through PARP1 to induce apoptosis. This mechanism shows a cell type-specific proapoptotic pathway that may lead to Notch agonist-based cancer therapeutics.

Topics: Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Cell Line, Tumor; Cell Separation; Child; Chromatin Immunoprecipitation; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Genes, Tumor Suppressor; Homeodomain Proteins; Humans; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Receptors, Notch; RNA, Messenger; Signal Transduction; Transcription Factor HES-1; Transfection

We explored the impact of mutations in the NOTCH1, FBW7 and PTEN genes on prognosis and downstream signaling in a well-defined cohort of 47 patients with pediatric T-cell acute lymphoblastic leukemia (T-ALL). In T-ALL lymphoblasts, we identified high-frequency mutations in NOTCH1 (n=16), FBW7 (n=5) and PTEN (n=26). NOTCH1 mutations resulted in 1.3- to 3.3-fold increased transactivation of an HES1 reporter construct over wild-type NOTCH1; mutant FBW7 resulted in further augmentation of reporter gene activity. NOTCH1 and FBW7 mutations were accompanied by increased median transcripts for NOTCH1 target genes (HES1, DELTEX1 and cMYC). However, none of these mutations were associated with treatment outcome. Elevated HES1, DELTEX1 and cMYC transcripts were associated with significant increases in transcript levels of several chemotherapy relevant genes, including MDR1, ABCC5, reduced folate carrier, asparagine synthetase, thiopurine methyltransferase, BCL2 and dihydrofolate reductase. PTEN transcripts positively correlated with HES1 and cMYC transcript levels. Our results suggest that (1) multiple factors should be considered with attempting to identify molecular-based prognostic factors for pediatric T-ALL, and (2) depending on the NOTCH1 signaling status, modifications in the types or dosing of standard chemotherapy drugs for T-ALL, or combinations of agents capable of targeting NOTCH1, AKT and/or mTOR with standard chemotherapy agents may be warranted.

Topics: Adolescent; Antineoplastic Combined Chemotherapy Protocols; Basic Helix-Loop-Helix Transcription Factors; Cell Cycle Proteins; Child; Child, Preschool; Cohort Studies; DNA Mutational Analysis; DNA, Neoplasm; F-Box Proteins; F-Box-WD Repeat-Containing Protein 7; Female; Gene Expression Regulation, Leukemic; Genes, Reporter; Homeodomain Proteins; Humans; Infant; Male; Mutation; Neoplasm Proteins; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma; Prognosis; PTEN Phosphohydrolase; Receptor, Notch1; Signal Transduction; Transcription Factor HES-1; Treatment Outcome; Ubiquitin-Protein Ligases; Young Adult