hes1-protein--human and Leukemia--Myeloid--Acute

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

Sirtuin 3 (SIRT3) deacetylase is a key regulator for chemoresistance in acute myeloid leukemia (AML) cells due to its capability of modulating mitochondrial metabolism and reactive oxygen species (ROS). SIRT3 is de-SUMOylated by SUMO-specific peptidase 1 (SENP1), which enhances its deacetylase activity. Therefore, dysregulation of SIRT3 SUMOylation may lead to fortified chemoresistance in AML. Indeed, SIRT3 de-SUMOylation was induced by chemotherapeutic agents, which in turn, exacerbated resistance against chemotherapies in AML by activating SIRT3 via preventing its proteasome degradation. Furthermore, RNA-seq revealed that expression of a collection of genes was altered by SIRT3 de-SUMOylation including inhibition of transcription factor Hes Family BHLH Transcription Factor 1 (HES1), a downstream substrate of Notch1 signaling pathway, leading to increased fatty acids oxidation (FAO). Moreover, the SENP1 inhibitor momordin-Ic or HES1 overexpression synergized with cytarabine to eradicate AML cells in vitro and in xenograft mouse models. In summary, the current study revealed a novel role of SIRT3 SUMOylation in the regulation of chemoresistance in AML via HES1-dependent FAO and provided a rationale for SIRT3 SUMOylation and FAO targeted interventions to improve chemotherapies in AML.

Topics: Animals; Drug Resistance, Neoplasm; Fatty Acids; Humans; Leukemia, Myeloid, Acute; Mice; Sirtuin 3; Sumoylation; 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

Acute myeloid leukemia (AML) is a group of heterogeneous hematopoietic malignancies sustained by leukemic stem cells (LSCs) that can resist treatment. Previously, we found that low expression of Hes1 was a poor prognostic factor for AML. However, the activation status of Hes1 and its regulation in LSCs and leukemic progenitors (LPs) as well as normal hematopoietic stem cells (HSCs) in Hes1-low AML patients have not been elucidated. In this study, the expression of Hes1 in LSCs and LPs was analyzed in adult CD34(+) Hes1-low AML with normal karyotype and the upstream microRNA (miRNA) regulators were screened. Our results showed that the level of either Hes1 or p21 was lower in LSCs or LPs than in HSCs whereas the level of miR-9 was highest in LPs and lowest in HSCs. An inverse correlation was observed in the expression of Hes1 and miR-9. Furthermore, we validated miR-9 as one of the regulators of Hes1 by reporter gene analysis. Knockdown of miR-9 by lentivirus infection suppressed the proliferation of AML cells by the induction of G0 arrest and apoptosis in vitro. Moreover, knockdown of miR-9 resulted in decreased circulating leukemic cell counts in peripheral blood and bone marrow, attenuated splenomegaly, and prolonged survival in a xenotransplant mouse model. Our results indicate that the miR-9 plays an important role in supporting AML cell growth and survival by downregulation of Hes1 and that miR-9 has potential as a therapeutic target for treating AML.

Topics: Adult; Aged; Animals; Antigens, CD34; Apoptosis; Biomarkers, Tumor; Blotting, Western; Cell Cycle; Cell Proliferation; Down-Regulation; Female; Follow-Up Studies; Hematopoietic Stem Cells; Humans; Karyotype; Leukemia, Myeloid, Acute; Male; Mice; Mice, Inbred NOD; Mice, SCID; MicroRNAs; Middle Aged; Neoplasm Staging; Neoplastic Stem Cells; Prognosis; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription Factor HES-1; Tumor Cells, Cultured; Xenograft Model Antitumor Assays; Young Adult

Although aberrant Notch activation contributes to leukemogenesis in T cells, the role of Notch pathway in acute myeloid leukemia (AML) remains controversial. To address this issue, we compared the expression levels of its downstream effector HES1 and p21 in bone marrow mononuclear cells (BMNCs) from 30 newly diagnosed AML patients and three AML cell lines to normal BMNCs. The results showed that both of them were downregulated in AML cells. In vitro, induced activation of HES1 by retrovirus in AML cell lines consistently led to AML cell growth arrest and apoptosis induction, which was associated with enhanced p21 expression. Furthermore, overexpression of HES1 in primary AML cells inhibited growth of AML in a xenograft mice model. In conclusion, we demonstrated the tumor suppressor role of HES1 in AML.

Topics: Adolescent; Adult; Animals; Basic Helix-Loop-Helix Transcription Factors; Child; Down-Regulation; Female; Gene Expression Regulation, Leukemic; Heterografts; HL-60 Cells; Homeodomain Proteins; Humans; Leukemia, Myeloid, Acute; Male; Mice; Mice, Inbred NOD; Mice, SCID; Middle Aged; Neoplasm Transplantation; Receptors, Notch; Transcription Factor HES-1; Tumor Suppressor Proteins; U937 Cells

To elucidate the impact of Hes1 on the proliferation and apoptosis of acute myeloid leukemia (AML) cells.. The expression levels of Hes1 and p21 in AML patient samples and myeloid leukemia cell lines were analyzed by real-time PCR. Hes1 was up-regulated by retrovirus transfection in AML cell lines and the proliferation capacity were assayed by MTT, cell cycle by Hoechst/PY, apoptosis by AnnexinV.. The expression of Hes1 in primary AML cells and HL-60, U937, KG1a cell lines were 0.67 ± 0.24, 0.59 ± 0.43, 0.42 ± 0.03, and 0.32 ± 0.26, respectively, and p21 were 0.54 ± 0.01, 0.44 ± 0.12, 0.36 ± 0.12, and 0.59 ± 0.43, respectively. Hes1 expression levels after transduction in HL-60, U937, KG1a were 4.9 ± 0.2, 5.2 ± 0.4, 5.8 ± 0.5, respectively. Induced activation of Hes1 led to AML cells growth arrest and apoptosis, which was associated with an enhanced p21 expression. Besides, activated Hes1 led to AML cells growth inhibition in vivo.. Hes1 could mediate growth arrest and apoptosis in AML cells, which may be a novel target for AML.

Topics: Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Cell Cycle; Cell Line, Tumor; Homeodomain Proteins; Humans; Leukemia, Myeloid, Acute; Transcription Factor HES-1; Up-Regulation

Although aberrant Notch activation contributes to leukemogenesis in T cells, its role in acute myelogenous leukemia (AML) remains unclear. Here, we report that human AML samples have robust expression of Notch receptors; however, Notch receptor activation and expression of downstream Notch targets are remarkably low, suggesting that Notch is present but not constitutively activated in human AML. The functional role of these Notch receptors in AML is not known. Induced activation through any of the Notch receptors (Notch1-4), or through the Notch target Hairy/Enhancer of Split 1 (HES1), consistently leads to AML growth arrest and caspase-dependent apoptosis, which are associated with B cell lymphoma 2 (BCL2) loss and enhanced p53/p21 expression. These effects were dependent on the HES1 repressor domain and were rescued through reexpression of BCL2. Importantly, activated Notch1, Notch2, and HES1 all led to inhibited AML growth in vivo, and Notch inhibition via dnMAML enhanced proliferation in vivo, thus revealing the physiological inhibition of AML growth in vivo in response to Notch signaling. As a novel therapeutic approach, we used a Notch agonist peptide that led to significant apoptosis in AML patient samples. In conclusion, we report consistent Notch-mediated growth arrest and apoptosis in human AML, and propose the development of Notch agonists as a potential therapeutic approach in AML.

Topics: Adolescent; Animals; Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Cell Line, Tumor; Cell Proliferation; Cell Survival; Child; DNA-Binding Proteins; Gene Expression; Homeodomain Proteins; Humans; Infant; Leukemia, Myeloid, Acute; Mice; Mice, Inbred NOD; Mice, Knockout; Mice, SCID; Mutation; Proto-Oncogene Proteins c-bcl-2; Receptors, Notch; RNA, Messenger; RNA, Neoplasm; Signal Transduction; Transcription Factor HES-1; Transcription Factors; Tumor Suppressor Protein p53