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Analysis of TGF-β signal transduction in patients with essential thrombocythemia

Research Project

Project/Area Number 11671007
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionSapporo Medical University

Principal Investigator

SAKAMAKI Sumio  SAPPORO MEDICAL UNIVERSITY, associate professor, 医学部, 助教授 (00196081)

Co-Investigator(Kenkyū-buntansha) NIITSU Yoshiro  Sapporo Medical University, professor, 医学部, 教授 (10045502)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
Keywordsmegakaryopoiesis / essential thrombocythemia / TGF-β / thrombopoietin / Smad / CFU-Meg / TaqMan PCR / gene transfection
Research Abstract

We recently reported that TGF-β is involved in negative feedback regulation of megakaryopoiesis in both normal volunteers (NV) and patients with Idiopathic Thrombocytopenic Purpura (Y.Hirayama et al. BLOOD,90,141,1998 ; S.Sakamaki et al. BLOOD,92,42,1999). In patients with Essential Thrombocythemia (ET), however, excess of megakaryopoiesis was observed despite the high concentration of TGF-β in BM, suggesting an escape mechanism for megakaryopoiesis from TGF-β action. To elucidate this mechanism, in this study, we analyzed TGF-β signal transduction pathways in megakaryocytes from ET and NV.Bone marrow mononuclear cells (BMMNC) (10^5 cells/dish) of ET and NV were cultured in α-MEM containing TGF-β (0, 0.1, 1.0 ng/mL), 100 ng/mL rh-TPO, 20% human fasting plasma and 0.8% methyl cellulose. After 14 days, CFU-Meg colony numbers were measured. RT-PCR of TGF-β receptor (type I, II) and Smad 2, 3, 4 and 5 were performed using total RNA prepared from CFU-Meg in liquid culture. TaqMan real-time RT-PCR were also employed to quantify the mRNA expression. The growth of CFU-Meg colonies derived from NV was completely suppressed by 1.0 ng/mL of TGF-β (mean inhibitory rate ; 92.7%) while those from ET patients (n=5) showed less suppression (mean inhibitory rate ; 24.1%) (p<0.05). Expressions of TGF-β receptor (type I, II) and Smad 2, 3 and 5 were equally detected in ET and NV.However, expression of Smad4 was significantly reduced in ET in comparison with those of NV (p<0.05). Transfection of Smad4 cDNA normalized the sensitivity of ET CFU-Meg to TGF-β. Thus, it was surmised that one of the mechanism for impaired sensitivity of CFU-Meg to TGF-β is the reduced expression of Smad4.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Sakamaki S, et al.: "Auto-antibodies against the specific epitope of human tropomyosin (s) detected by a peptide-based enzyme immunoassay in sera o patients with ulcerative colitis show antibody dependent cell-mediated cytotoxicity against HLA-DPwg transfected L-cells."Gut. 47. 236-241 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Matsunaga T,Sakamaki S, et al.: "GST-p gene transduced hematopoietic progenitor cell transplantation overcomes the bone maroow toxicity of cyclophosphamide in mice"Hum Gene Ther. 11(2). 1679-1689 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sato Y,Sakamaki S, et al.: "In vivo gene delivery to tumor cells by transferrin-streptavidin-DNA conjugate."FASEB J. 14. 2108-2118 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mezawa S,Sakamaki S, et al.: "A study of carboplatin coated tube for the unresectable cholangiocarcinoma."Hepatology,. 32(5). 916-923 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kohda K,Sakamaki S, et al.: "Long-term survival and late-onset complications of cancer patients treated with high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation."Int J Hematol. 73(2). 251-257 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Lu Y,Sakamaki S, et al.: "Prevention of lethal acute-graft-versus-host disease in mice by oral administration of T helper 1 inhibitor-TAK-603."Blood. 97. 1123-1130 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Matsunaga T, Sakamaki S, Ishigaki S, Kohda K, Takeda M, Kato J, Kuroda H, Hirayama Y, Kusakabe T, Akiyama T, Kuga T, Niitsu Y, Masaoka T, Sagawa T, Matsumoto Y.: "Use of serum in diagnosing and monitoring cytomegalovirus reactivation in bone marrow transplant recipients."Int J Hematol. 69. 105-111 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sakamaki S, Hirayama Y, Matsunaga T, Kuroda H, Kusakabe T, Akiyama T, Konuma Y, Sasaki K, Tsuji N, Okamoto T, Kobune M, Kogawa K, Kato J, Takimoto R, Koyama R, Niitsu Y.: "Transforming growth factor-β1 (TGF-β1) induces thrombopoietin from bone marrow stromal cells, which stimulates the expression of TGF-β receptor on magakaryocytes and, in turn renders them susceptible to suppression by TGF-β itself with high specificity."Blood. 94(6). 1961-1970 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sakamaki S, Takayanagi N, Yoshizaki N, Hayashi S, Takayama T, Kato J, Kogawa K, Yamauchi N, Tsuji N, Nobuoka A, Ayabe T, Kohgo Y, Niitsu Y.: "Auto-antibodies against the specific epitope of human tropomyosin (s) detected by a peptide-based enzyme immunoassay in sera of patients with ulcerative colitis show antibody dependent cell-mediated cytotoxicity anainst HLA-DPw9 transfected L-cells."Gut. 47. 236-241 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Matsunaga T, Sakamaki S, Kuga T, Kuroda H, Kusakabe T, Akiyama T, Konuma Y, Hirayama Y, Kobune M, Kato J, Sasaki K, Kogawa K, Koyama R, Niitsu Y.: "GST-π gene transduced hematopoietic progenitor cell transplantation overcomes the bone maroow toxcity of cyclophosphamide in mice."Hum Gene Ther. 11(12). 1679-1689 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sato Y, Yamauchi N, Takahashi M, Sasaki K, Fukaura J, Neda H, Jujii S, Hirayama M, Itoh Y, Koshita Y, Kogawa K, Kato J, Sakamaki S, Niitsu Y.: "In vivo gene delivery to tumor cells by transferrin-streptavidin-DNA conjugate."FASEB J. 14. 2108-2118 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Lu Y, Sakamaki S, Kuroda H, Kusakabe T, Konuma Y, Akiyama T, Fujimi A, Nishiie K, Matsunaga T, Hirayama Y, Kato J, Kon S, Kogawa K, Niitsu Y.: "Prevention of lethal acute-graft-versus-host disease in mice by oral administration of T helper 1 inhibitor-TAK-603."Blood. 97. 1123-1130 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sakamaki S, et al.: "Auto-antibodies against the specific apitope of human tropomyosin (s) detacted by a peptide-based enzyme imnunoassay in sera of pationts with ulcerative colitis show atibody dependent cell-mediated cytotoxicity against HLA-DPw9 transfected L-cells."Gut. 47. 236-241 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Matsunaga T,sakamaki S, et al.: "GST-p gene transduced hematopoietic progenitor cell transplantation overcomes the bone maroow toxicity of cyclophosphamide in mice"Hum Gene Ther. 11(12). 1679-1689 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sato Y, Sakamaki S, et al.: "In vivo gene delivery to tumor cells by transferrin-streptavidin-DNA conjugate."FASEB J. 14. 2108-2118 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mezawa S,Sakamaki S, et al.: "A study of carboplatin coated tube for the unresectable cholangiocarcinoma."Hepatology,. 32(5). 916-923 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kohda K,Sakamaki S, et al.: "Long-term survival and late-onset complications of cancer patients treated with high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation."Int J Hematol. 73(2). 251-257 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Lu Y,Sakamaki S, et al.: "Prevention of lethal acute-graft-versus-host disease in mice by oral administration of T helper 1 inhibitor-TAK-603."Blood. 97. 1123-1130 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Matsunaga T, Sakamaki S, et al.: "Use of PCR serum in diagnosing and monitoring cytomegalovirus reactivation in bone marrow transplant recipients."Int J Hematol.. 69. 105-111 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Teramoto J, Sakamaki S, et al.: "Systemic lupus erythematosus with a giant rectal ulcer and perforation"Internal Medicine.. 38. 643-649 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kogawa K, Sakamaki S, et al.: "Enhanced inhibition of experimental metastasis by the combination chemotherapy of Cu-Zn SOD and adriamycin.Clin Exp Metastasis."Clin Exp Metastasis.. 17. 239-244 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sakamaki S, Hirayama Y, et al.: "Transforming growth factor-β1(TGF-β1)induces thrombopoietin from bone marrow stromal cells, which stimulates the expression of TGF-β receptor on megakaryocytes and, in tum, renders them susceptible to suppression by TGF-β itself with high specificity."Blood.. 94. 1961-1970 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nobuoka A, Sakamaki S, et al.: "A case of malignant lymphoma producing autoantibody against platelet glycoprotein Ib."Int J Hematology.. 70. 200-206 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sakamaki S, Takayanagi N, et al.: "Auto-antibodies against the specific epitope of human tropomyosin(s) detected by a peptide-based enzyme immunoassay in sera of patients with ulcerative colitis show antibody dependent cell-mediated cytotoxicity against HLA-DAw9 transfected L-cells."Gut. (in press).

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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