1996 Fiscal Year Annual Research Report

KITレセプターの突然変異によるマスト細胞腫の発生機構

Research Project

Project/Area Number	07670245
Research Institution	Osaka University
Principal Investigator	辻村亨大阪大学, 医学部・附属病院, 助手 (20227408)
Co-Investigator(Kenkyū-buntansha)	北村幸彦大阪大学, 医学部, 教授 (70028520) 岡部勝大阪大学, 微生物研究所・感染動物実験施設, 助教授 (30089875)
Keywords	KITレセプターチロシンキナーゼ / 機能獲得性突然変異 / マスト細胞腫 / トランスジェニックマウス / レトロウイルスベクター
Research Abstract	c-kitレセプター・チロシンキナーゼ(KIT)には機能獲得性突然変異(キナーゼ領域のアルパラギン酸をバリン或いはチロシンへ置換する点突然変異)が見出されている。インターロイキン3(IL-3)に依存して増殖する培養マスト細胞株IC-2はマスト細胞としての多くの表現型を保つがKITを発現していない。IC-2細胞にアルパラギン酸をバリンへ置換した変異c-kit遺伝子(活性型c-kit遺伝子)をレトロウイルス発現系を用いて導入すると、導入細胞はIL-3やKITのリガンドであるstem cell factor(SCF)を含まない培地でも活発に増殖するようになりヌードマウスに移植すると腫瘍を形成した後白血病化した。また、正常マウスの骨髄細胞に活性型c-kit遺伝子を含むレトロウイルスを感染させると、感染骨髄細胞から造血因子がなくてもメチルセルロースを含む培地で顆粒球マクロファージコロニーばかりでなくマスト細胞コロニーや混合コロニーが出現し、感染骨髄細胞をマウスに移植すると白血病が発症した。主要組織適合抗原クラスIのプロモーターで発現制御される活性型c-kit遺伝子を導入したトランスジェニックマウスでは白血病やリンパ腫が発症した。更に、自律性に増殖する培養マスト細胞腫株FMA3ではKITにアルパラギン酸を他のアミノ酸へ置換する変異を認めなかったが傍細胞膜領域に7個のアミノ酸欠失を認め、この欠失のためにKITがSCFの結合なしに構成的に二量体を形成し活発化することがわかった。この欠失型KITをコードするc-kit遺伝子をレトロウイルス発現系を用いてIL-3依存性IC-2細胞へ導入すると、導入細胞は自律性に増殖するようになりヌードマウスに移植すると腫瘍を形成した後白血病化した。本研究により突然変異により構成的に活性化するKITは生物学的活性をもつことが明らかとなり、活性型KITはマスト細胞の腫瘍化に直接的に関与すると考えられる。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Hashimoto,K: "Transforming and differentiation-inducing potentials of constitutively activated c-kit mutant genes in the IC-2 murine interleukin-3-dependent mast cell line" Am J Pathol. 148・1. 189-200 (1996)
[Publications] Tsujimura,T: "Constitutive activation of c-kit in FMA3 murine mastocytoma cells caused by deletion of seven amino acids at the juxtamembrane domain" Blood. 87・1. 273-283 (1996)
[Publications] Moriyama,Y: "Role of aspartic acid 814 in the function and expression of c-kit receptor tyrosine kinase" J Biol Chem. 271・7. 3347-3350 (1996)
[Publications] Kitayama,H: "Neoplastic transformation of normal hematopoietic cells by constitutively activating mutations of c-kit receptor tyrosine kinase" Blood. 88・3. 995-1004 (1996)
[Publications] Tsujimura,T: "Involvement of transcription factor encoded by the mi locus in the expression of c-kit receptor tyrosine kinase in cultured mast cells of mice" Blood. 88・4. 1225-1233 (1996)
[Publications] Morii,E: "Regulation of mouse mast cell protease 6 gene expression by transcription factor encoded by the mi locus" Blood. 88・7. 2488-2494 (1996)
[Publications] Kanakura,Y: "The c-kit receptor tyrosine kinase : its roles in hematological malignancies" Cancer J. 9・2. 79-82 (1996)
[Publications] Tsujimura,T: "Role of c-kit receptor tyrosine kinase for development,survival and neoplastic transformation of mast cells" Pathol Int. 46・12. 933-938 (1996)
[Publications] Adachi,S: "Inhibition of attachment between cultured mast cells and fibroblasts by phorbol 12-myristate 13-acetate and stem cell factor" Exp Hematol. 23・1. 58-65 (1995)
[Publications] Kamada,S: "bcl-2 deficiency in mice leads to pleiotropic anbormalities : Accelerated lymphoid cell death in thymus and spleen,polycystic kidney,hair hypopigmentation,and distorted small intestine" Cancer Res. 55・2. 354-359 (1995)
[Publications] Kitayama,H: "Constitutive activating mutations of c-kit receptor tyrosine kinase cnfer factor-independent growth and tumorigenicity of factor-dependent hematopoietic cell lines" Blood. 85・3. 790-798 (1995)
[Publications] Tsujimura T: "Substitution of an aspartic acid results in constitutive activation of c-kit receptor tyrosine kinase in a rat tumor mast cell line RBL-2H3" Int Arch Allergy Immunol. 106・4. 377-385 (1995)
[Publications] Matsumura,I: "Growth response of acute myeloblastic leukemia cells to recombinant human thrombopoietin" Blood. 86・2. 703-709 (1995)
[Publications] Kanno,H: "Primary structure of murine red cell-type pyruvate kinase (PK) and molecular characterization of PK deficiency identified in the CBA strain" Blood. 86・8. 3205-3210 (1995)
[Publications] Morimoto,M: "Pyruvate kinase deficiency of mice associated with nonpherocytic hemolytic anemia and cure of the anemia by marrow transplantation without host irradiation" Blood. 86・11. 4323-4330 (1995)
[Publications] Kitamura,Y: "Regulation of development,survival and neoplastic growth of mast cells through c-kit receptor" Int Arch Allergy Immunol. 107・1〜3. 54-56 (1995)
[Publications] Kitamura,Y: "Hematopoietic Lineages : Regulation of cell production and development" marcel dekker,New York, in press
[Publications] Kitamura,Y: "Biological and Molecular Aspects of Mast Cell and Basophil Differentiation and Function" Reven Press,New York, 270 (1995)

1996 Fiscal Year Annual Research Report

KITレセプターの突然変異によるマスト細胞腫の発生機構

Principal Investigator

辻村 亨 大阪大学, 医学部・附属病院, 助手 (20227408)

Research Products

[Publications] Hashimoto,K: "Transforming and differentiation-inducing potentials of constitutively activated c-kit mutant genes in the IC-2 murine interleukin-3-dependent mast cell line" Am J Pathol. 148・1. 189-200 (1996)

[Publications] Tsujimura,T: "Constitutive activation of c-kit in FMA3 murine mastocytoma cells caused by deletion of seven amino acids at the juxtamembrane domain" Blood. 87・1. 273-283 (1996)

[Publications] Moriyama,Y: "Role of aspartic acid 814 in the function and expression of c-kit receptor tyrosine kinase" J Biol Chem. 271・7. 3347-3350 (1996)

[Publications] Kitayama,H: "Neoplastic transformation of normal hematopoietic cells by constitutively activating mutations of c-kit receptor tyrosine kinase" Blood. 88・3. 995-1004 (1996)

[Publications] Tsujimura,T: "Involvement of transcription factor encoded by the mi locus in the expression of c-kit receptor tyrosine kinase in cultured mast cells of mice" Blood. 88・4. 1225-1233 (1996)

[Publications] Morii,E: "Regulation of mouse mast cell protease 6 gene expression by transcription factor encoded by the mi locus" Blood. 88・7. 2488-2494 (1996)

[Publications] Kanakura,Y: "The c-kit receptor tyrosine kinase : its roles in hematological malignancies" Cancer J. 9・2. 79-82 (1996)

[Publications] Tsujimura,T: "Role of c-kit receptor tyrosine kinase for development,survival and neoplastic transformation of mast cells" Pathol Int. 46・12. 933-938 (1996)

[Publications] Adachi,S: "Inhibition of attachment between cultured mast cells and fibroblasts by phorbol 12-myristate 13-acetate and stem cell factor" Exp Hematol. 23・1. 58-65 (1995)

[Publications] Kamada,S: "bcl-2 deficiency in mice leads to pleiotropic anbormalities : Accelerated lymphoid cell death in thymus and spleen,polycystic kidney,hair hypopigmentation,and distorted small intestine" Cancer Res. 55・2. 354-359 (1995)

[Publications] Kitayama,H: "Constitutive activating mutations of c-kit receptor tyrosine kinase cnfer factor-independent growth and tumorigenicity of factor-dependent hematopoietic cell lines" Blood. 85・3. 790-798 (1995)

[Publications] Tsujimura T: "Substitution of an aspartic acid results in constitutive activation of c-kit receptor tyrosine kinase in a rat tumor mast cell line RBL-2H3" Int Arch Allergy Immunol. 106・4. 377-385 (1995)

[Publications] Matsumura,I: "Growth response of acute myeloblastic leukemia cells to recombinant human thrombopoietin" Blood. 86・2. 703-709 (1995)

[Publications] Kanno,H: "Primary structure of murine red cell-type pyruvate kinase (PK) and molecular characterization of PK deficiency identified in the CBA strain" Blood. 86・8. 3205-3210 (1995)

[Publications] Morimoto,M: "Pyruvate kinase deficiency of mice associated with nonpherocytic hemolytic anemia and cure of the anemia by marrow transplantation without host irradiation" Blood. 86・11. 4323-4330 (1995)

[Publications] Kitamura,Y: "Regulation of development,survival and neoplastic growth of mast cells through c-kit receptor" Int Arch Allergy Immunol. 107・1〜3. 54-56 (1995)

[Publications] Kitamura,Y: "Hematopoietic Lineages : Regulation of cell production and development" marcel dekker,New York, in press

[Publications] Kitamura,Y: "Biological and Molecular Aspects of Mast Cell and Basophil Differentiation and Function" Reven Press,New York, 270 (1995)

辻村亨大阪大学, 医学部・附属病院, 助手 (20227408)