2003 Fiscal Year Annual Research Report

造血幹細胞におけるテロメラーゼ活性の制御機構の解析と制御分子の単離の試み

Research Project

Project/Area Number	14657245
Research Institution	Osaka University
Principal Investigator	金倉譲大阪大学, 医学系研究科, 教授 (20177489)
Co-Investigator(Kenkyū-buntansha)	柴山浩彦大阪大学, 医学系研究科, 助手 (60346202) 水木満佐央大阪大学, 医学系研究科, 講師 (80283761) 松村到大阪大学, 医学系研究科, 助教授 (00294083)
Keywords	テロメラーゼ / 造血幹細胞 / c-Myc / 自己複製 / 不死化 / Notch / HOXB4
Research Abstract	テロメアとは染色体末端部分に存在する(TTAGGG)nの繰り返し配列である。DNA合成の際に再生されず、細胞分裂を繰り返すと短縮していき、細胞分裂の回数を規定している。造血幹細胞は自己複製能と多分化能を有し、個体の生存期間、造血幹細胞集団を維持しつつ血液細胞を供給するが、この細胞集団は一般の正常体細胞とは異なり、テロメア配列を補充するテロメラーゼ活性を有している。本研究においては、造血幹細脚の自己複製・不死化機構とその際のテロメラーゼ活性の制御機構について解析を行い、以下の結果を見出した。 1.マウス骨髄より単離したLin^-Sca-1^+の造血幹細胞に活性型NotchやHOXB4を導入すると、自己複製がもたらされ、この際にはテロメラーゼ活性の上昇が認められた。 2.Notch、HOXB4による自己複製の際の各種細胞周期制御分子の発現変化を調べたところ、G1/S期に関わる細胞周期制御分子の中でcyclin D1やE2F1などと比較してc-mycの発現上昇が最も早期から認められた。また、両分子はルシフェラーゼアッセイにおいてC-mycプロモーターを活性化した。 3.マウス造血幹細胞に4-hydrxytamoxifen(4-HT)によってc-Mycの活性が誘導可能なc-Mycと変異型エストロゲン受容体(ERT)のキメラ分子c-Myc/ERTを導入したところ、c-Myc/ERTは造血幹細胞に自己複製をもたらし、テロメラーゼ活性を上昇させた。以上の結果から、Notch、HOXB4による造血幹細胞の自己複製、テロメラーゼ活性の制御において、c-Mycが中心的な役割を担うと考えられた(J.Biol.Chem.,in revision)。

Research Products

(41 results)

All Other

All Publications (41 results)

[Publications] Tanaka H, et al.: "E2F-1 and c-Myc potentiate apoptosis through inhibition NF-κB that facilitates MnSOD-mediated ROS elimination"Molecular Cell. 9. 1017-1029 (2002)
[Publications] Ezoe S, et al.: "GATA-2/estrogen receptor chimera regulates cytokine-dependentgrowth of hematopoiesis through accumulation of p21WAF1 and p27Kip1 proteins"Blood. 100. 3512-3520 (2002)
[Publications] Ueda S, et al.: "Critical roles of c-kit tyrosine residues 567 and 719 in stem cell cellfactor-induced chemotaxis : contribution of src family kinase and PI3-kinase on calcium mobilization and cell migration"Blood. 100. 3342-3349 (2002)
[Publications] Sonoyama J, et al.: "Functional cooperation among Ras, STAT5 and PI3-K is required for fulloncogenic activities of BCR/ABL in K562 cells"J.Biol.Chem.. 277. 8076-8082 (2002)
[Publications] Nishimura Ji, et al.: "Long-term support of hematopoiesis by a single stem cell clone inpatients with paroxysmal nocturnal hemoglobinuria"Blood. 99. 2748-2751 (2002)
[Publications] Nojima J, et al.: "Acquired activated protein C resistance, associated with anti-protein Santibody as a strong risk factor for DVT in non-SLE patients"Thromb.Haemost.. 88. 716-722 (2002)
[Publications] Nojima J, et al.: "Acquired activated protein C resistance is associated with the co-existence of anti-prothrombin antibodies and lupus anticoagulant activity in patients with systemic lupus erythematosus"Br.J.Haematol.. 118. 577-583 (2002)
[Publications] Matsumura I, et al.: "Molecular control of megakaryopoiesis and thrombopoiesis"Int.J.Hematol.. 75. 473-483 (2002)
[Publications] Ueda S, et al.: "Constitutive activation of c-kit by the juxtamembrane but not thecatalytic domain mutations is inhibited selectively by tyrosine kinase inhibitors STI571 and AG1296"Int.J.Hematol.. 76. 427-435 (2002)
[Publications] Kabutomori O, et al.: "Movement of toxic granulation neutrophils and C-reactive protein ininflammatory processes"Am.J.Med.. 112. 595-596 (2002)
[Publications] Kabutomori O, et al.: "Induction of toxic granulation in neutrophils by granulocyte colony-stimulating factor"Eur.J.Haematol.. 69. 187-188 (2002)
[Publications] Yamanishi H, et al.: "Modification of fully automated total iron-binding capacity (TIBC) assayin serum and comparison with dimension TIBC method"Thromb.Haemost.. 88. 716-722 (2002)
[Publications] Yamanishi H, et al.: "Relation between iron content of serum ferritin and clinical status factorsextracted by factor analysis in patients with hypefferritinemia"Clin.Biochem.. 35. 523-529 (2002)
[Publications] Muller-Tidow C, et al.: "Mutations of growth factor receptor Flt3 in acute myeloid leukemia : transformation of myeloid cells by Ras-dependent and Ras-independentmechanisms"Dtsch.Med.Wochenschr.. 127. 2195-2200 (2002)
[Publications] Yang Q, et al.: "Hepatocyte nuclear factor-1a modulates pancreatic β-cell growth byregulating the expression of insulin-like growth factor-1 in INS-1 cells"Diabetes. 51. 1785-1792 (2002)
[Publications] Nanbu-Wakao, R, et al.: "Stimulation of 3T3-L1 adipogenesis by signal transducer and activator of transcription 5"Mol.Endocrinol.. 16. 1565-1576 (2002)
[Publications] Imai Y, et al.: "High prevalence of HCV infedtion in patients with B-cell non-Hodgkin's lymphoma : comparision with birth cohort- and sex-matched blood donors in a Japanese population"Hepatology. 35. 974-976 (2002)
[Publications] Kuratsune H, et al.: "Brain regions involved in fatigue sensation : reduced acetylcarnitine uptake into the brain"Neuroimage. 17. 1256-1265 (2002)
[Publications] Hashimoto K, et al.: "Necessity of tyrosine 719 and phosphatidylinositol 3'-kinase-mediated signal pathway inconstitutive activation and oncogenic potential of c-kit receptor tyrosine kinase with the D814V mutation"Blood. 101. 1094-1102 (2003)
[Publications] Kawasaki A, et al.: "Opposing effects of PML and PML/RAR α on STAT3 activity"Blood. 101. 3668-3673 (2003)
[Publications] Mizuki M, et al.: "Suppression of myeloid transcription factors and induction of STAT response genes by AML-specific Flt3 mutations"Blood. 101. 3164-3173 (2003)
[Publications] Kataoka Y, et al.: "Reciprocal inhibition between MyoD and STAT3 in the regulation of growth and differentiation of myoblasts"J.BIol.Chem.. 278. 44178-44187 (2003)
[Publications] Saeki Y, et al.: "Enhanced production of osteopontin in multiple myeloma : clinical and pathogenicimplications"Br.J.Haematol.. 123. 263-270 (2003)
[Publications] Zhang X, et al.: "Constitutively activated Rho GTPases regulate the growth and morphology of hairy cellleukemia (HCL) cells"Int.J.Hematology. 77. 263-273 (2003)
[Publications] Takakuwa T, et al.: "Establishment and characterization of unique cell lines derived from pyothorax-associated lymphoma which develops in long-standing pyothorax and is stronglyassociated with Epstein-Barr virus infection"Cancer Sci.. 94. 858-863 (2003)
[Publications] Matsumura I, et al.: "Molecular mechanisms of E2F1- and c-Myc-enhanced apoptosis"Rec.Res.Dev.in Mol.& Cell.Biol.. 4. 311-324 (2003)
[Publications] Matsumura I, et al.: "Roles for E2F1 and c-Myc in the regulation of cell growth and survival"Cell Cycle. 2. 333-338 (2003)
[Publications] Mizuki M, et al.: "Constitutive active receptor tyrosine kinase in hematological neoplasia"Rec.Res.Dev.in Mol.& Cell.Biol.. 4. 85-100 (2003)
[Publications] Mizuki M, et al.: "Oncogenic receptor tyrosine kinase in leukemia"Cellular and Molecular Biology. 49. 907-922 (2003)
[Publications] Matsumura I, et al.: "Cell cycle regulation in hematopoietic stem cells"Res.Adv.in Blood. 2. 39-49 (2003)
[Publications] Okawa Y, et al.: "Purification of N-terminally truncated histone H2A-monoubiquitin conjugates from leukemic cell nuclei : probable proteolytic products of ubiquitinated H2A"Int.J.Biochem.Cell Biol.. 35. 1588-1600 (2003)
[Publications] Yamanishi H, et al.: "Total iron-binding capacity calculated from serum transferrin concentration or serumiron concentration and unsaturated iron-binding capacity"Clin.Chem.. 49. 175-178 (2003)
[Publications] Yamanishi H, et al.: "Modification of the colorimetric assay for serum unsaturated iron-binding capacity"Clin.Chem.. 49. 1023-1025 (2003)
[Publications] Kimura S, et al.: "Enzymatic assay for determination of bicarbonate ion in plasma using urea amidolyase"Clin.Chim.Acta.. 328. 179-184 (2003)
[Publications] Kimura S, et al.: "New enzymatic assay for serum urea nitrogen using urea amidolyase"J.Clin.Lab.Anal.. 17. 52-56 (2003)
[Publications] Kabutomori O, et al.: "Marked decreases of total and immature reticulocytes in myelodysplastic syndromeamong patients with pancytopenia"Acta Haematol.. 109. 212-213 (2003)
[Publications] Kabutomori O, et al.: "Incorrect measurement of leukocyte counts in post-bone marrow transplantation (P-BMT) patients"Ann.Hematol.. 82. 529-530 (2003)
[Publications] Ueda Y, et al.: "New homogeneous HDL-cholesterol assay without the influence of high TG sample using the selective detergent to lipoproteins"J Clin Lab Anal.. 17. 201-208 (2003)
[Publications] Tanaka S, et al.: "Autoantibodies against muscarinic cholinergic receptor in chronic fatigue syndrome"Int.J.Mol.Med.. 12. 225-230 (2003)
[Publications] Shibayama H, et al.: "Identification of a cytokine-induced antiapoptotic molecule Anamorsin essential for definitive hematopoiesis"J.Exp.Med.. 199. 581-592 (2004)
[Publications] Ezoe S, et al.: "Roles for CDK inhibtors in hematopoietic system"Cell Cycle. (in press). (2004)

2003 Fiscal Year Annual Research Report

造血幹細胞におけるテロメラーゼ活性の制御機構の解析と制御分子の単離の試み

Principal Investigator

金倉 譲 大阪大学, 医学系研究科, 教授 (20177489)

Research Products

[Publications] Tanaka H, et al.: "E2F-1 and c-Myc potentiate apoptosis through inhibition NF-κB that facilitates MnSOD-mediated ROS elimination"Molecular Cell. 9. 1017-1029 (2002)

[Publications] Ezoe S, et al.: "GATA-2/estrogen receptor chimera regulates cytokine-dependentgrowth of hematopoiesis through accumulation of p21WAF1 and p27Kip1 proteins"Blood. 100. 3512-3520 (2002)

[Publications] Ueda S, et al.: "Critical roles of c-kit tyrosine residues 567 and 719 in stem cell cellfactor-induced chemotaxis : contribution of src family kinase and PI3-kinase on calcium mobilization and cell migration"Blood. 100. 3342-3349 (2002)

[Publications] Sonoyama J, et al.: "Functional cooperation among Ras, STAT5 and PI3-K is required for fulloncogenic activities of BCR/ABL in K562 cells"J.Biol.Chem.. 277. 8076-8082 (2002)

[Publications] Nishimura Ji, et al.: "Long-term support of hematopoiesis by a single stem cell clone inpatients with paroxysmal nocturnal hemoglobinuria"Blood. 99. 2748-2751 (2002)

[Publications] Nojima J, et al.: "Acquired activated protein C resistance, associated with anti-protein Santibody as a strong risk factor for DVT in non-SLE patients"Thromb.Haemost.. 88. 716-722 (2002)

[Publications] Nojima J, et al.: "Acquired activated protein C resistance is associated with the co-existence of anti-prothrombin antibodies and lupus anticoagulant activity in patients with systemic lupus erythematosus"Br.J.Haematol.. 118. 577-583 (2002)

[Publications] Matsumura I, et al.: "Molecular control of megakaryopoiesis and thrombopoiesis"Int.J.Hematol.. 75. 473-483 (2002)

[Publications] Ueda S, et al.: "Constitutive activation of c-kit by the juxtamembrane but not thecatalytic domain mutations is inhibited selectively by tyrosine kinase inhibitors STI571 and AG1296"Int.J.Hematol.. 76. 427-435 (2002)

[Publications] Kabutomori O, et al.: "Movement of toxic granulation neutrophils and C-reactive protein ininflammatory processes"Am.J.Med.. 112. 595-596 (2002)

[Publications] Kabutomori O, et al.: "Induction of toxic granulation in neutrophils by granulocyte colony-stimulating factor"Eur.J.Haematol.. 69. 187-188 (2002)

[Publications] Yamanishi H, et al.: "Modification of fully automated total iron-binding capacity (TIBC) assayin serum and comparison with dimension TIBC method"Thromb.Haemost.. 88. 716-722 (2002)

[Publications] Yamanishi H, et al.: "Relation between iron content of serum ferritin and clinical status factorsextracted by factor analysis in patients with hypefferritinemia"Clin.Biochem.. 35. 523-529 (2002)

[Publications] Muller-Tidow C, et al.: "Mutations of growth factor receptor Flt3 in acute myeloid leukemia : transformation of myeloid cells by Ras-dependent and Ras-independentmechanisms"Dtsch.Med.Wochenschr.. 127. 2195-2200 (2002)

[Publications] Yang Q, et al.: "Hepatocyte nuclear factor-1a modulates pancreatic β-cell growth byregulating the expression of insulin-like growth factor-1 in INS-1 cells"Diabetes. 51. 1785-1792 (2002)

[Publications] Nanbu-Wakao, R, et al.: "Stimulation of 3T3-L1 adipogenesis by signal transducer and activator of transcription 5"Mol.Endocrinol.. 16. 1565-1576 (2002)

[Publications] Imai Y, et al.: "High prevalence of HCV infedtion in patients with B-cell non-Hodgkin's lymphoma : comparision with birth cohort- and sex-matched blood donors in a Japanese population"Hepatology. 35. 974-976 (2002)

[Publications] Kuratsune H, et al.: "Brain regions involved in fatigue sensation : reduced acetylcarnitine uptake into the brain"Neuroimage. 17. 1256-1265 (2002)

[Publications] Hashimoto K, et al.: "Necessity of tyrosine 719 and phosphatidylinositol 3'-kinase-mediated signal pathway inconstitutive activation and oncogenic potential of c-kit receptor tyrosine kinase with the D814V mutation"Blood. 101. 1094-1102 (2003)

[Publications] Kawasaki A, et al.: "Opposing effects of PML and PML/RAR α on STAT3 activity"Blood. 101. 3668-3673 (2003)

[Publications] Mizuki M, et al.: "Suppression of myeloid transcription factors and induction of STAT response genes by AML-specific Flt3 mutations"Blood. 101. 3164-3173 (2003)

[Publications] Kataoka Y, et al.: "Reciprocal inhibition between MyoD and STAT3 in the regulation of growth and differentiation of myoblasts"J.BIol.Chem.. 278. 44178-44187 (2003)

[Publications] Saeki Y, et al.: "Enhanced production of osteopontin in multiple myeloma : clinical and pathogenicimplications"Br.J.Haematol.. 123. 263-270 (2003)

[Publications] Zhang X, et al.: "Constitutively activated Rho GTPases regulate the growth and morphology of hairy cellleukemia (HCL) cells"Int.J.Hematology. 77. 263-273 (2003)

[Publications] Takakuwa T, et al.: "Establishment and characterization of unique cell lines derived from pyothorax-associated lymphoma which develops in long-standing pyothorax and is stronglyassociated with Epstein-Barr virus infection"Cancer Sci.. 94. 858-863 (2003)

[Publications] Matsumura I, et al.: "Molecular mechanisms of E2F1- and c-Myc-enhanced apoptosis"Rec.Res.Dev.in Mol.& Cell.Biol.. 4. 311-324 (2003)

[Publications] Matsumura I, et al.: "Roles for E2F1 and c-Myc in the regulation of cell growth and survival"Cell Cycle. 2. 333-338 (2003)

[Publications] Mizuki M, et al.: "Constitutive active receptor tyrosine kinase in hematological neoplasia"Rec.Res.Dev.in Mol.& Cell.Biol.. 4. 85-100 (2003)

[Publications] Mizuki M, et al.: "Oncogenic receptor tyrosine kinase in leukemia"Cellular and Molecular Biology. 49. 907-922 (2003)

[Publications] Matsumura I, et al.: "Cell cycle regulation in hematopoietic stem cells"Res.Adv.in Blood. 2. 39-49 (2003)

[Publications] Okawa Y, et al.: "Purification of N-terminally truncated histone H2A-monoubiquitin conjugates from leukemic cell nuclei : probable proteolytic products of ubiquitinated H2A"Int.J.Biochem.Cell Biol.. 35. 1588-1600 (2003)

[Publications] Yamanishi H, et al.: "Total iron-binding capacity calculated from serum transferrin concentration or serumiron concentration and unsaturated iron-binding capacity"Clin.Chem.. 49. 175-178 (2003)

[Publications] Yamanishi H, et al.: "Modification of the colorimetric assay for serum unsaturated iron-binding capacity"Clin.Chem.. 49. 1023-1025 (2003)

[Publications] Kimura S, et al.: "Enzymatic assay for determination of bicarbonate ion in plasma using urea amidolyase"Clin.Chim.Acta.. 328. 179-184 (2003)

[Publications] Kimura S, et al.: "New enzymatic assay for serum urea nitrogen using urea amidolyase"J.Clin.Lab.Anal.. 17. 52-56 (2003)

[Publications] Kabutomori O, et al.: "Marked decreases of total and immature reticulocytes in myelodysplastic syndromeamong patients with pancytopenia"Acta Haematol.. 109. 212-213 (2003)

[Publications] Kabutomori O, et al.: "Incorrect measurement of leukocyte counts in post-bone marrow transplantation (P-BMT) patients"Ann.Hematol.. 82. 529-530 (2003)

[Publications] Ueda Y, et al.: "New homogeneous HDL-cholesterol assay without the influence of high TG sample using the selective detergent to lipoproteins"J Clin Lab Anal.. 17. 201-208 (2003)

[Publications] Tanaka S, et al.: "Autoantibodies against muscarinic cholinergic receptor in chronic fatigue syndrome"Int.J.Mol.Med.. 12. 225-230 (2003)

[Publications] Shibayama H, et al.: "Identification of a cytokine-induced antiapoptotic molecule Anamorsin essential for definitive hematopoiesis"J.Exp.Med.. 199. 581-592 (2004)

[Publications] Ezoe S, et al.: "Roles for CDK inhibtors in hematopoietic system"Cell Cycle. (in press). (2004)

金倉譲大阪大学, 医学系研究科, 教授 (20177489)