ABSTRACT
The 8;21 translocation is a common chromosomal abnormality in acute myeloid leukemia (AML). We recently identified a naturally occurring leukemogenic splice variant, AML1-ETO9a (acute myeloid leukemia-1 transcription factor and the eight-twenty-one corepressor-9a), of t(8;21). To understand the leukemic potential of AML1-ETO9a, we performed microarray analysis with the murine multipotential hematopoietic FDCP-mix A4 cell line. We identified changes in expression of various genes including CD44. CD44 is a type I transmembrane protein and functions as the major cellular adhesion molecule for hyaluronic acid, a component of the extracellular matrix. CD44 is expressed in most human cell types and is implicated in myeloid leukemia pathogenesis. We show that the presence of AML1-ETO9a significantly increased the expression of CD44 at both RNA and protein levels. Furthermore, the CD44 promoter is bound by AML1-ETO9a and AML1-ETO at the chromatin level. In addition, in the AML1-ETO9a leukemia mouse model CD44 is regulated in a cell context-dependent manner. Thus, our observations suggest that AML1-ETO and its splice variant AML1-ETO9a are able to regulate the expression of the CD44 gene, linking the 8;21 translocation to the regulation of a cell adhesion molecule that is involved in the growth and maintenance of the AML blast/stem cells.
Subject(s)
Chromosomes, Human, Pair 21 , Chromosomes, Human, Pair 8 , Hyaluronan Receptors/genetics , Leukemia, Myeloid/genetics , Translocation, Genetic , Acute Disease , Alternative Splicing , Animals , Cell Differentiation , Cell Division , Cell Survival , Core Binding Factor Alpha 2 Subunit/genetics , Gene Expression Regulation, Leukemic , Humans , Hyaluronan Receptors/metabolism , K562 Cells , Leukemia, Myeloid/pathology , Mice , Neoplastic Stem Cells/cytology , Neoplastic Stem Cells/physiology , Oligonucleotide Array Sequence Analysis , Oncogene Proteins, Fusion/genetics , RUNX1 Translocation Partner 1 ProteinABSTRACT
Macrophage mannose receptor (MMR) recognizes the pattern of carbohydrates exposed on microorganisms and mediates endocytosis in macrophages. We have synthesized glycoconjugate cationic polymers carrying 3,6-branched alpha-D-mannoside, a trimannose conjugate (TMC) with a high affinity for mannose-specific lectins. Culture with 10 microM TMC for 6 hours induced adhesion and aggregation in NKM-1, a human myelomonocytic leukemia cell line. TMC also stimulated the accumulation of fluorescein isothiocyanate (FITC)-dextran (FITC-DX). This accumulation seemed to be mediated by endocytosis via MMR because mannan, which specifically binds to MMR, inhibited FITC-DX accumulation. Expression of CD14, adhesion molecules, and costimulatory molecules was induced for 24 hours in NKM-1 and in fresh leukemia blasts from 4 patients with acute myeloid leukemia (AML) M4 and M5 subtypes (French-American-British classification). To clarify the binding mechanism, we compared mannose conjugates and a monomer of mannose regarding their effects on endocytosis and enhancement of CD14 and CD86 expression. A polymer of monomannose clusters with a lower affinity for lectins slightly stimulated exdocytosis, whereas a monomer of trimannose had no effect. These findings suggest that concatenation between MMR and TMC may play an important role in the activation of monocytic leukemia cells. TMC may become a good candidate to target MMRs of leukemia cells.
Subject(s)
Adjuvants, Immunologic/metabolism , Endocytosis/drug effects , Lectins, C-Type , Leukemia, Myeloid/pathology , Mannose-Binding Lectins , Mannose/pharmacology , Trisaccharides/pharmacology , Acute Disease , Antigens, CD/drug effects , Antigens, CD/metabolism , B7-2 Antigen , Bone Marrow Cells/drug effects , Bone Marrow Cells/pathology , Cell Adhesion/drug effects , Dextrans/pharmacokinetics , Fluorescein-5-isothiocyanate/analogs & derivatives , Fluorescein-5-isothiocyanate/pharmacokinetics , Humans , Immunophenotyping , Leukemia, Myeloid/immunology , Lipopolysaccharide Receptors/drug effects , Lipopolysaccharide Receptors/metabolism , Macrophages/chemistry , Mannose/analogs & derivatives , Mannose/chemical synthesis , Mannose Receptor , Membrane Glycoproteins/drug effects , Membrane Glycoproteins/metabolism , Molecular Structure , Receptors, Cell Surface/metabolism , Receptors, Cell Surface/physiology , Trisaccharides/chemical synthesis , Trisaccharides/metabolism , Tumor Cells, Cultured/drug effectsABSTRACT
Effects of prolonged noradrenaline infusion on the density of cardiac beta-adrenoceptors, phosphodiesterase (PDE) and adenylate cyclase (AC) activities, and the ability of NKH477, 6-(3-dimethylaminopropionyl) forskolin hydrochloride, to increase tension development and heart rate were studied in rat cardiac preparations. Noradrenaline infusion (400 micrograms/kg/hr, s.c.) for 7 days significantly decreased cardiac beta-adrenoceptor density (Bmax), whereas the binding affinity (Kd) of the ligand was unchanged. The basal cardiac PDE activity was increased in treated rats, whereas there was no difference in the basal cardiac AC activity between treated and untreated rats. Significant decreases in basal developed tension and heart rate were observed in the left and right atrial muscles from treated rats, respectively. The positive inotropic and chronotropic potencies of NKH477 were unaffected by noradrenaline infusion, whereas the positive inotropic potencies of isoproterenol and 3-isobutyl-1-methylxanthine were significantly reduced. Thus, NKH477 appears to be superior to beta-adrenoceptor agnosits or PDE inhibitors as a cardiotonic drug in the treatment of heart failure accompanied by beta-adrenoceptor downregulation.
