Glyoxalase-I is a novel target against Bcr-Abl+ leukemic cells acquiring stem-like characteristics in a hypoxic environment

M Takeuchi; S Kimura; J Kuroda; E Ashihara; M Kawatani; H Osada; K Umezawa; E Yasui; M Imoto; T Tsuruo; A Yokota; R Tanaka; R Nagao; T Nakahata; Y Fujiyama; T Maekawa

doi:10.1038/cdd.2010.6

Glo1 inhibitors for neuropsychiatric and anti-epileptic drug development

Biochemical Society Transactions ◽

10.1042/bst20140027 ◽

2014 ◽

Vol 42 (2) ◽

pp. 461-467 ◽

Cited By ~ 10

Author(s):

Katherine M.J. McMurray ◽

Margaret G. Distler ◽

Preetpal S. Sidhu ◽

James M. Cook ◽

Leggy A. Arnold ◽

...

Keyword(s):

Therapeutic Potential ◽

Partial Agonist ◽

Neuropsychiatric Disorders ◽

Anxiety And Depression ◽

Glyoxalase I ◽

Pharmacological Treatments ◽

Delayed Onset ◽

Anti Epileptic Drug ◽

Novel Target ◽

Target Effects

Many current pharmacological treatments for neuropsychiatric disorders, such as anxiety and depression, are limited by a delayed onset of therapeutic effect, adverse side effects, abuse potential or lack of efficacy in many patients. These off-target effects highlight the need to identify novel mechanisms and targets for treatment. Recently, modulation of Glo1 (glyoxalase I) activity was shown to regulate anxiety-like behaviour and seizure-susceptibility in mice. These effects are likely to be mediated through the regulation of MG (methylglyoxal) by Glo1, as MG acts as a competitive partial agonist at GABAA (γ-aminobutyric acid A) receptors. Thus modulation of MG by Glo1 represents a novel target for treatment. In the present article, we evaluate the therapeutic potential of indirectly modulating MG concentrations through Glo1 inhibitors for the treatment of neuropsychiatric disorders.

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Glyoxalase-I: a novel target for pancreatic cancer?

Pancreatology ◽

10.1016/j.pan.2018.05.390 ◽

2018 ◽

Vol 18 (4) ◽

pp. S145

Author(s):

Marcus Hollenbach ◽

Jana Lorenz ◽

Ines Sommerer ◽

Sebastian Sonnenberg ◽

Joachim Mössner ◽

...

Keyword(s):

Pancreatic Cancer ◽

Glyoxalase I ◽

Novel Target

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Calcineurin/NFAT Pathway Is a Novel Target of Therapeutic Interest in Lymphoid Malignancies.

Blood ◽

10.1182/blood.v108.11.1827.1827 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1827-1827

Author(s):

Hind Medyouf ◽

Helene Alcade ◽

Caroline Berthier ◽

Anne Janin ◽

Hugues de Thé ◽

...

Keyword(s):

T Cell ◽

Transgenic Mice ◽

Mouse Models ◽

T Cell Development ◽

Signalling Pathway ◽

Leukemic Cells ◽

Lymphoid Malignancies ◽

Nfat Activation ◽

Novel Target ◽

And Function

Abstract Calcineurin (PP2B) is an ubiquitously expressed serine/threonine protein phosphatase dependent on calcium calmodulin. Calcineurin plays a pivotal role in many important biological processes, including the development and function of the immune system. Sustained signalling through calcium/calcineurin results in the activation of transcription factors of the NFAT family (Nuclear Factor of Activated T cells). Mouse genetic studies have demonstrated a strong epistatic relationship between calcineurin and NFAT activation and function in many developmental processes. In lymphoid cells, the calcineurin/NFAT signalling pathway is essential to specific aspects of T cell development and plays a central role in the activation of the immune response and in its homeostatic control. Despite their central role in T cell development, immune function and homeostasis, deregulation of calcineurin/NFAT pathway have not been shown so far to have a pro-oncogenic role in lymphomagenesis. Using our TEL-JAK2 transgenic mouse model of T-ALL, we investigated the activation status and importance of the calcineurin/NFAT signalling pathway in leukemogenesis. We found that the calcineurin/NFAT pathway is activated in leukemic cells of TEL-JAK2 transgenic mice. Western blot and EMSA experiments showed that all NFAT members expressed in the lymphoid lineage (NFAT1, 2 and 4) are found in an active hypophosphorylated form, they are located in the nucleus, and exhibit an increase in their DNA binding activity. In order to investigate whether constitutive calcineurin/NFAT activity was specific to TEL-JAK2 leukemia or whether it is a more general property of leukemic cells, we analyzed NFAT activation in leukemic cells of transgenic mice aberrantly expressing oncoproteins involved in lymphoma in human. This showed that calcineurin/NFATpathway is not only aberantly active in leukemic cells of TEL-JAK2 transgenic animals, but also in other mouse models of human leukemia. We also found that the activation of the calcineurin/NFAT pathway is only observed when cells are rapidly analyzed following their explantation and dissociation from diseased animals. Indeed, if cells are incubated in tissue culture for a short period of time (60–120min.), essentially stochiometric rephosphorylation, meaning inactivation, of NFAT is observed. This indicates that activation of the calcineurin/NFAT pathway depends upon signals generated in the in vivo tumor cell environment and is not -or not solely- under the control of the initiating oncogene. Using these mouse models we conducted pre-clinical studies that indicated that calcineurin/NFAT pathway is an attractive novel target of therapeutic interest in lymphoid malignancies.

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Glyoxalase-I Inhibition Eliminates Hypoxia-Adapted CML Cells Resistant to Imatinib, Dasatinib, and INNO-406

Blood ◽

10.1182/blood.v112.11.3185.3185 ◽

2008 ◽

Vol 112 (11) ◽

pp. 3185-3185

Author(s):

Miki Takeuchi ◽

Shinya Kimura ◽

Junya Kuroda ◽

Takashi Ushiki ◽

Makoto Kawatani ◽

...

Keyword(s):

Stem Cells ◽

Enzymatic Activity ◽

Cell Lines ◽

K562 Cells ◽

Adaptation To Hypoxia ◽

Glyoxalase I ◽

Hematopoietic Stem ◽

Hypoxic Conditions ◽

Hypoxic Environment ◽

Nog Mice

Abstract The development of ABL tyrosine kinase inhibitor (TKI), imatinib mesylate and second-generation ABL TKIs such as dasatinib and nilotinib have dramatically improved the efficacy of chronic myeloid leukemia (CML) treatment, but “total cell kill” may not be possible with TKIs alone because they do not appear to eliminate CML stem cells. Although the CML stem cell niche has not yet been clearly identified, it may be a hypoxic environment, like that of normal hematopoietic stem cells, which mainly exists in the low oxygen-consuming region in the bone marrow (BM). Notably, most cancer cells that adapt to hypoxia are resistant to a variety of cell-death stimuli. Thus, we hypothesized that CML cells, in particular CML stem cells, adapt to hypoxia for their persistence. To test this notion, we first examined the oxygenated status of CML cells in vivo by transplanting K562 cells into NOD/SCID/gcnull (NOG) mice. Six weeks later, pimonidazole, which specifically accumulates in tissues with <1.3% O2, was administered and the mice were sacrificed. The K562 cells had mainly engrafted into the epiphysis, which is presumed to be a particularly hypoxic environment, and these engrafted cells were positively stained by antibody to pimonidazole. This suggests that the engrafted leukemic cells survived in hypoxic conditions. To investigate the role that adaptation to hypoxia plays in CML-cell persistence, we generated two hypoxia-adapted cell lines that can survive in hypoxic conditions and identified their unique characteristics compared to their parental cells. For this, the CML-derived cell lines K562, KCL-22, BV173 and MYL were cultivated in 1.0% O2 for two weeks. Two subclones that survived were obtained from K562 and KCL22 and were denoted as K562/HA and KCL22/HA, respectively. Both cell lines proliferated continuously for months in 1.0% O2 conditions in RPMI1640 medium containing 10% FCS but lacking any additional nutrient supplies, although their growth was slower than that of their parental counterparts under 20% oxygen in the same media. We found that, compared to their parental counterparts, K562/HA and KCL22/HA had higher glyoxalase-I (Glo-I) expression and kinase activity. Glo-I is an enzyme that detoxifies methylglyoxal, a cytotoxic byproduct of glycolysis. Interestingly, BCR-ABL phosphorylation was suppressed in both hypoxia-adapted cell lines. However, the phosphorylation of AKT, STAT and ERK was unchanged in these hypoxia-adapted cells, which suggests the presence of alternative hypoxia-specific activation pathways. When NOG mice were inoculated with equivalent numbers of K562/HA and K562 cells, K562/HA engrafted more rapidly than K562 and K562/HA-engrafted mice died earlier than K562-engrafted mice. Thus, adaptation to hypoxia is crucial for the engraftment and proliferation of CML cells in the BM. Notably, compared to their parental counterparts, K562/HA and KCL22/HA cells were less sensitive to TKIs (imatinib, dasatinib, and INNO-406) and alkylating agents (daunorubicin and busulfan). However, they were more sensitive to Glo-I inhibitors such as S-p-bromobenzylglutathione cyclopentyl diester (BBGC), 2-crotonyloxymethyl- 4,5,6-trihydroxycylohex-2-enone (COTC), and methyl-gerfelin. Thus, Glo-I enzymatic activity plays an important role in CML-cell adaptation to hypoxic environments, which means Glo-1 may be an alternative molecular target for therapies aiming to eliminate those CML cells that adapt to hypoxia and thereby acquire resistance to conventional chemotherapies, including those based on TKIs. Indeed, when K562/HA-transplanted mice were treated with 100mg/kg/day BBGC for 8 days or left untreated, the treated mice survived longer than the untreated mice. In contrast, the survival of mice transplanted with the parental cell line K562 did not change whether they were treated with BBGC or left untreated. Thus, Glo-1 enzymatic activity plays an important role in CML-cell survival under hypoxic environmental conditions and Glo-I inhibitors could be promising agents for the elimination of CML (stem) cells that persist in the hypoxic BM milieu

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Targeting RIOK2 ATPase activity leads to decreased protein synthesis and cell death in acute myeloid leukemia

Blood ◽

10.1182/blood.2021012629 ◽

2021 ◽

Author(s):

Jan-Erik Messling ◽

Karl Agger ◽

Kasper L Andersen ◽

Kristina Kromer ◽

Hanna Maria Kuepper ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Protein Synthesis ◽

Myeloid Leukemia ◽

Leukemic Cell ◽

Leukemic Cells ◽

Proof Of Concept ◽

Molecule Inhibitor ◽

Novel Target ◽

Acute Myeloid

Novel therapies for the treatment of acute myeloid leukemia (AML) are urgently needed as current treatments do not cure the majority of AML patients. Here, we report on a domain-focused, kinome-wide CRISPR-Cas9 screen to identify protein kinase targets for the treatment of AML, which led to the identification of Rio-kinase 2 (RIOK2) as a potential novel target. We show that loss of RIOK2 leads to a decrease in protein synthesis and to ribosomal instability followed by apoptosis in leukemic cells, but not in fibroblasts. Moreover, we demonstrate that the ATPase function of RIOK2 is required for cell survival. By using a small molecule inhibitor, we show that pharmacological inhibition of RIOK2 similarly leads to loss of protein synthesis and apoptosis and affects leukemic cell growth in vivo. Our results provide proof-of-concept for targeting RIOK2 as a potential treatment for AML patients.

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Glyoxalase I: A novel target for neuroendocrine tumors?

Pancreatology ◽

10.1016/j.pan.2021.05.169 ◽

2021 ◽

Vol 21 ◽

pp. S64

Author(s):

M. Hollenbach ◽

M. Tatschner ◽

A. Hoffmeister ◽

P. Michl ◽

S. Krug

Keyword(s):

Neuroendocrine Tumors ◽

Glyoxalase I ◽

Novel Target

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Glyoxalase-I Is a Potential Therapeutic Target for Chronic Myelogenous Leukemia Cells That Have Acquired Drug Resistance through Adaptation to Hypoxia.

Blood ◽

10.1182/blood.v110.11.2931.2931 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2931-2931

Author(s):

Miki Takeuchi ◽

Junya Kuroda ◽

Shinya Kimura ◽

Eishi Ashihara ◽

Kazuo Umezawa ◽

...

Keyword(s):

Drug Resistance ◽

Cell Death ◽

Cell Lines ◽

Chronic Myelogenous Leukemia ◽

Myelogenous Leukemia ◽

Leukemic Cells ◽

Leukemia Cells ◽

Adaptation To Hypoxia ◽

Glyoxalase I ◽

Hypoxic Conditions

Abstract Growth under hypoxic conditions can lead to resistance to a variety of cell death stimuli in cancer cells. Bone marrow (BM) has less vasculature than other organs, and the rapid growth of leukemic cells in BM frequently outpaces the oxygen supply. Thus, adaptation to hypoxia may be critical to the survival and proliferation of leukemic cells. However, the role of hypoxia in the treatment of leukemia has received little attention. We hypothesized that leukemic cells acquire drug resistance by adapting to hypoxia in the BM, and that identifying the underlying molecular mechanisms for this adaptation would reveal new approaches to the development of new therapeutic targets in drug- resistant leukemia cells. To test this hypothesis, we cultured five chronic myelogenous leukemia (CML)-derived cell lines (BV173, K562, KCL22, KT-1, and MYL) continuously in 1.0% oxygen. Among them, hypoxia resistant (HR) subclones of K562 and KCL22 cells (K562/HR and KCL22/HR) were selected under 1.0% oxygen conditions, and these cells were able to grow for several months and to be suspended in RPMI1640 plus 10% FCS without any additional nutrients, although their growth rate was slower than parental cells in 20% oxygen. Both K562/HR and KCL22/HR cells produced less ATP (78% and 92%, respectively), perhaps due to impairment of the oxygen-dependent energy production pathway, and HR cells showed higher expression and enhanced kinetics of glyoxalase-I (Glo-I), a detoxification kinase of methylglyoxal (a cytotoxic byproduct of glycolysis), compared to parental cells. Glo-I has previously been implicated in drug resistance in cancer, including leukemia. Autophosphorylation of Bcr-Abl was reduced in HR cell lines, while the phosphorylation status of the downstream signalling molecules Akt and Stat5 was unchanged, which suggests that there are alternative, hypoxia-specific activators of these pathways. K562/HR and KCL22/HR cells were less sensitive to chemotherapeutic agents such as the Bcr-Abl tyrosine kinase (TK) inhibitors imatinib, dasatinib, nilotinib, and INNO-406, and the alkylating agents daunorucbicin and busulfan. In contrast, K562/HR and KCL22/HR cells were more sensitive to cell death induced by a Glo-I inhibitor, COTC, compared to parental cells. These results indicated that Glo-I plays an important role in survival under hypoxic conditions, and that Glo- I may be an alternative molecular target in CML cells that have acquired resistance to conventional chemotherapeutics through adaptating to hypoxia.

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Role of PKC isozymes in water transport in toad urinary bladder

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100085812 ◽

1991 ◽

Vol 49 ◽

pp. 302-303

Author(s):

A.J. Mia ◽

L.X. Oakford ◽

T. Yorio

Keyword(s):

Urinary Bladder ◽

Apical Membrane ◽

Membrane Surface ◽

Protein A ◽

Cell Types ◽

Leukemic Cells ◽

Toad Urinary Bladder ◽

Pkc Isozymes ◽

Antibody Labeling

Protein kinase C (PKC) isozymes, when activated, are translocated to particulate membrane fractions for transport to the apical membrane surface in a variety of cell types. Evidence of PKC translocation was demonstrated in human megakaryoblastic leukemic cells, and in cardiac myocytes and fibroblasts, using FTTC immunofluorescent antibody labeling techniques. Recently, we reported immunogold localizations of PKC subtypes I and II in toad urinary bladder epithelia, following 60 min stimulation with Mezerein (MZ), a PKC activator, or antidiuretic hormone (ADH). Localization of isozyme subtypes I and n was carried out in separate grids using specific monoclonal antibodies with subsequent labeling with 20nm protein A-gold probes. Each PKC subtype was found to be distributed singularly and in discrete isolated patches in the cytosol as well as in the apical membrane domains. To determine if the PKC isozymes co-localized within the cell, a double immunogold labeling technique using single grids was utilized.

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