1999 Fiscal Year Annual Research Report

神経変性疾患と悪性腫瘍の分子医学

Research Project

Project/Area Number	10CE2006
Research Institution	Nagoya University
Principal Investigator	高橋雅英名古屋大学, 医学部, 教授 (40183446)
Co-Investigator(Kenkyū-buntansha)	古川鋼一名古屋大学, 医学部, 教授 (80211530) 祖父江元名古屋大学, 医学部, 教授 (20148315) 村松喬名古屋大学, 医学部, 教授 (00030891) 浜口道成名古屋大学, 医学部, 教授 (90135351)
Keywords	神経変性疾患 / 悪性腫瘍 / RET遺伝子 / ミッドカイン / 糖鎖合成酵素遺伝子 / Src遺伝子 / トランスジェニックマウス / ノックアウトマウス
Research Abstract	1.神経変性疾患、神経機能に関わる分子の同定と機能解析 (1)分子インデックス法を用いて、筋萎縮性側索硬化症(ALS)の脊髄前角細胞と正常の脊髄前角細胞において発現量の大きく変化している遺伝子群を検索した。その結果、84個の遺伝子フラグメントを得、うち2つについては胎児脳cDNAライブラリーより完全長のcDNAをクローニングし、塩基配列を決定した。(祖父江) (2)CAGリピート病の神経培養細胞モデルにおいて、Hsp40,Hsp70により神経細胞死、封入体形成が抑制できることを明らかにした。(祖父江) (3)ヒト神経芽細胞腫細胞を用いて神経栄養因子GDNFにより発現誘導される遺伝子群をDifferential Display法により解析した。現在までに16個の誘導遺伝子を同定し、そのうち10個が既知の遺伝子で6個が未知の遺伝子であった。未知の遺伝子については全長のcDNAをクローニングし、塩基配列を決定中である。(高橋) (4)神経細胞の表面蛋白でミッドカイン(MK)と強く結合し、受容体候補となる高分子量蛋白を同定し、大量精製後、部分アミノ酸配列を決定した。(村松) (5)MKにアポトーシス防止能があることを見出し、その活性にはPI3キナーゼが関与していることを明らかにした。またMKがコンドロイチン硫酸プロテオグリカンと強く結合することを明らかにした。(村松) (6)糖脂質糖鎖の神経機能における役割を明らかにする目的で、4種類の新規の糖脂質糖鎖合成酵素遺伝子cDNAをクローニングし、その遺伝子構造と発現パターンを明らかにした。(古川) 2.癌の増殖、浸潤、転移の分子機序とその制御 (1)WDリピート構造を有する新規のシグナル伝達因子MAWD遺伝子を同定し、この遺伝子を強制発現すると細胞が癌化すること、ヒト乳癌組織では癌組織特異的にMAWD蛋白質の発現が亢進していることを明らかにした。(浜口) (2)v-Src癌化細胞のマトリックスメタロプロテアーゼの分泌亢進と浸潤の活性化は、Ras-MEK1-MAPキナーゼ系のシグナル伝達を必要とすることを明らかにした。(浜口) (3)Differential Display法により多発性内分泌腫瘍症2A型、2B型変異を有するRET蛋白によって発現誘導される遺伝子を同定した。その中には既知の遺伝子が5個、未知の遺伝子が6個含まれていた。(高橋) (4)G2/M期を制御するプロテインキナーゼAuroraが細胞周期においてAPC-ユピキチン-プロテアソーム系によリ分解される機構を解明した。(吉川) 3.神経変性疾患、悪性腫瘍のモデル動物の作製と利用 (1)多発性内分泌腫瘍症 2A型変異RET遺伝子導入トランスジェニックマウスを作製した。このトランスジェニックマウスでは甲状腺髄様癌が100%発症し、in vitroでの実験結果と同様にRET蛋白の二量体形成が腫瘍化に重要であることが判明した。(高橋) (2)トリプレットリピート病の1つである球脊髄性筋萎縮症(SBMA)型の異常を有するアンドロゲンレセプター遺伝子を導入したトランスジェニックマウスを4系統確立し、マウス運動能力低下、神経細胞における核内封入体形成を認めた。(祖父江) (3)GM2/GD2合成酵素遺伝子、GD3合成酵素遺伝子ノックアウトマウスおよびGM2/GD2合成酵素遺伝子のトランスジェニックマウスの解析により、複合型ガングリオシドが神経系組織の維持と再生に必須であることが判明した。(古川) 4.効率的な遺伝子治療法の確立と利用 (1)Ring-finger構造を有するRFP遺伝子のプロモーター領域を解析し、RFPプロモーターがヒトの細胞にて非常に高い活性を有することを明らかにした。このプロモーターが遺伝子治療用の発現ベクターに応用可能かどうか現在検討中である。(高橋)

Research Products
(25 results)

All Other

All Publications (25 results)

[Publications] Hideki Murakami: "Rho-dependent and-independent tyrosine phosphorylation of focal adhesion kinase, paxillin and p130Cas mediated by Ret kinase."Oncogene. 18. 1975-1982 (1999)
[Publications] Toshihide Iwashita: "Biological and biochemical properties of Ret with kinase domain mutations identified in multiple endocrine neoplasia type 2B and familial medullary thyroid carcinoma."Oncogene. 18. 3919-3922 (1999)
[Publications] Hideki Murakami: "Enhanced phosphatidylinositol 3-kinase activity and high phosphorylation state of its downstream signalling molecules mediated by Ret with the MEN 2B mutation."Biochem.Biophys.Res.Commun.. 262. 68-75 (1999)
[Publications] Kou Miyazaki: "Crirical amino acid substitutions in the Src SH3 domain that convert c-Src to be oncogenic."Biochem.Biophys.Res.Commun.. 263. 759-764 (1999)
[Publications] Yoshihiro Ishiguro: "The role of amino acids surrounding tyrosine 1062 in Ret in specific binding of the Shc phosphotyrosine binding domain."Endocrinology. 140. 3992-3998 (1999)
[Publications] Susumu Ohta: "Midkine is expressed during repair of bone fracture and promotes chondrogenesis."J.Bone Miner.Res.. 14. 1132-1144 (1999)
[Publications] Chunlin Ye: "Expression of midkine in the early stage of carcinogenesis in human colorectal cancer."Brit.J.Cancer. 79. 179-184 (1999)
[Publications] Satoshi Fukumoto: "Expression cloning of mouse cDNA of CMP-NeuAc : lactosylceramide α2, 3-sialyltransferase (GM3 synthase), an enzyme that initiates the synthesis of gangliosides. "J.Biol.Chem.. 274. 9271-9276 (1999)
[Publications] Nobuaki Maeda: "A receptor-like protein tyrosine phosphatase (PTPζ/RPTP β) binds a heparin binding growth factor midkine : Involvement of arginine 78 of midkine in the high affinity binding to PTP ζ."J.Biol.Chem.. 274. 12474-79 (1999)
[Publications] Jinmin Zhao: "Attenuation of the interleukin 2 signals in the spleen cells of complex ganglioside-lacking mice."J.Biol.Chem.. 274. 13744-47 (1999)
[Publications] Tetsuya Okajima: "Human homolog of Caenorhabditis elegans sqv-3 gene is galactosyl-transferase I involved in the biosynthesis of the glycosaminoglycan-protein linkage region of proteoglycans."J.Biol.Chem.. 274. 22915-18 (1999)
[Publications] Anwarul A.Akhand: "Nitric oxide controls Src kinase activity through a redox-based molecular modification."J.Biol.Chem.. 274. 25821-26 (1999)
[Publications] Tetsuya Okajima: "Molecular basis for the progeroid variant of Ehlers-Danlos syndrome : Identification and characterization of two mutations in galactosyltrans-ferase I gene."J.Biol.Chem.. 274. 28841-44 (1999)
[Publications] Fumiaki Tanaka: "Tissue-specific somatic mosaicism in spinal and bulbar muscular atrophy is dependent on CAG-repeat length and Androgen receptor-gene expression level."Am.J.Hum.Genet.. 65. 966-973 (1999)
[Publications] Kenichiro Misu: "Late onset familial amyloid polyneuropathy type I (transthyretin Met-30-associated familial amyloid polyneuropathy) unrelated to andemic focus in Japan.Clinicopathologial and genetic features"Brain. 122. 1951-1962 (1999)
[Publications] Toshiya Sato: "Transgenic mice harboring a full-length human mutant DRPLA gene exhibit age-dependent intergenerational and somatic instabilities of CAG repeats comparable with those in DRPLA patients."Hum.Mol.Genet.. 8. 99-106 (1999)
[Publications] Shingo Yasuda: "Triggering of neuronal cell death by accumulation of activated SEK1 on nuclear polyglutamine aggregations in PML bodies."Genes to Cells. 294. (1999)
[Publications] Aye Aye Thant: "Ras pathway is required for the activation of MMP-2 and for the invasion of src-transformed 3Y1."Oncogene. 18. 6555-6563 (1999)
[Publications] Takeshi Senga: "Clustered cysteine residues in the kinase domain of v-Src : critical role for protein stability, cell transformation and sensitivity to herbimycin A."Oncogene. 19. 273-279 (2000)
[Publications] Satoru Matsuda: "Molecular cloning and characterization of human MAWD, a novel protein containing WD-40 repeats frequently overexpressed in breast cancer."Cancer Res.. 60. 13-17 (2000)
[Publications] Masashi Kato: "Ultraviolet light induces redox reaction-mediated dimerization and superactivation of oncogenic ret tyrosine kinases."Mol.Biol.Cell. 11. 93-101 (2000)
[Publications] Kazuhiro Ishiguro: "Syndecan-4 deficiency impairs focal adhesion formation only under restricted conditions."J.Biol.Chem.. 275. 5249-5252 (2000)
[Publications] Yasushi Kobayashi: "Chaperones, Hsp70 and Hsp40, suppress aggregate formation and apoptosis in cultured neuronal cells expressing truncated androgen receptor protein with expanded polyglutamine."J.Biol.Chem.. (in press).
[Publications] Satoshi Fukumoto: "GD3 synthase gene expression in PC12 cells results in the continuous activation of TrkA and ERK1/2 and enhanced proliferation."J.Biol.Chem.. (in press).
[Publications] Mitsuru Horiba: "Neointima formation in a restenosis model is suppressed in midkine-deficient mice,"J.Clin.Invest.. (in press).

1999 Fiscal Year Annual Research Report

神経変性疾患と悪性腫瘍の分子医学

Principal Investigator

高橋 雅英 名古屋大学, 医学部, 教授 (40183446)

Research Products

[Publications] Hideki Murakami: "Rho-dependent and-independent tyrosine phosphorylation of focal adhesion kinase, paxillin and p130Cas mediated by Ret kinase."Oncogene. 18. 1975-1982 (1999)

[Publications] Toshihide Iwashita: "Biological and biochemical properties of Ret with kinase domain mutations identified in multiple endocrine neoplasia type 2B and familial medullary thyroid carcinoma."Oncogene. 18. 3919-3922 (1999)

[Publications] Hideki Murakami: "Enhanced phosphatidylinositol 3-kinase activity and high phosphorylation state of its downstream signalling molecules mediated by Ret with the MEN 2B mutation."Biochem.Biophys.Res.Commun.. 262. 68-75 (1999)

[Publications] Kou Miyazaki: "Crirical amino acid substitutions in the Src SH3 domain that convert c-Src to be oncogenic."Biochem.Biophys.Res.Commun.. 263. 759-764 (1999)

[Publications] Yoshihiro Ishiguro: "The role of amino acids surrounding tyrosine 1062 in Ret in specific binding of the Shc phosphotyrosine binding domain."Endocrinology. 140. 3992-3998 (1999)

[Publications] Susumu Ohta: "Midkine is expressed during repair of bone fracture and promotes chondrogenesis."J.Bone Miner.Res.. 14. 1132-1144 (1999)

[Publications] Chunlin Ye: "Expression of midkine in the early stage of carcinogenesis in human colorectal cancer."Brit.J.Cancer. 79. 179-184 (1999)

[Publications] Satoshi Fukumoto: "Expression cloning of mouse cDNA of CMP-NeuAc : lactosylceramide α2, 3-sialyltransferase (GM3 synthase), an enzyme that initiates the synthesis of gangliosides. "J.Biol.Chem.. 274. 9271-9276 (1999)

[Publications] Nobuaki Maeda: "A receptor-like protein tyrosine phosphatase (PTPζ/RPTP β) binds a heparin binding growth factor midkine : Involvement of arginine 78 of midkine in the high affinity binding to PTP ζ."J.Biol.Chem.. 274. 12474-79 (1999)

[Publications] Jinmin Zhao: "Attenuation of the interleukin 2 signals in the spleen cells of complex ganglioside-lacking mice."J.Biol.Chem.. 274. 13744-47 (1999)

[Publications] Tetsuya Okajima: "Human homolog of Caenorhabditis elegans sqv-3 gene is galactosyl-transferase I involved in the biosynthesis of the glycosaminoglycan-protein linkage region of proteoglycans."J.Biol.Chem.. 274. 22915-18 (1999)

[Publications] Anwarul A.Akhand: "Nitric oxide controls Src kinase activity through a redox-based molecular modification."J.Biol.Chem.. 274. 25821-26 (1999)

[Publications] Tetsuya Okajima: "Molecular basis for the progeroid variant of Ehlers-Danlos syndrome : Identification and characterization of two mutations in galactosyltrans-ferase I gene."J.Biol.Chem.. 274. 28841-44 (1999)

[Publications] Fumiaki Tanaka: "Tissue-specific somatic mosaicism in spinal and bulbar muscular atrophy is dependent on CAG-repeat length and Androgen receptor-gene expression level."Am.J.Hum.Genet.. 65. 966-973 (1999)

[Publications] Kenichiro Misu: "Late onset familial amyloid polyneuropathy type I (transthyretin Met-30-associated familial amyloid polyneuropathy) unrelated to andemic focus in Japan.Clinicopathologial and genetic features"Brain. 122. 1951-1962 (1999)

[Publications] Toshiya Sato: "Transgenic mice harboring a full-length human mutant DRPLA gene exhibit age-dependent intergenerational and somatic instabilities of CAG repeats comparable with those in DRPLA patients."Hum.Mol.Genet.. 8. 99-106 (1999)

[Publications] Shingo Yasuda: "Triggering of neuronal cell death by accumulation of activated SEK1 on nuclear polyglutamine aggregations in PML bodies."Genes to Cells. 294. (1999)

[Publications] Aye Aye Thant: "Ras pathway is required for the activation of MMP-2 and for the invasion of src-transformed 3Y1."Oncogene. 18. 6555-6563 (1999)

[Publications] Takeshi Senga: "Clustered cysteine residues in the kinase domain of v-Src : critical role for protein stability, cell transformation and sensitivity to herbimycin A."Oncogene. 19. 273-279 (2000)

[Publications] Satoru Matsuda: "Molecular cloning and characterization of human MAWD, a novel protein containing WD-40 repeats frequently overexpressed in breast cancer."Cancer Res.. 60. 13-17 (2000)

[Publications] Masashi Kato: "Ultraviolet light induces redox reaction-mediated dimerization and superactivation of oncogenic ret tyrosine kinases."Mol.Biol.Cell. 11. 93-101 (2000)

[Publications] Kazuhiro Ishiguro: "Syndecan-4 deficiency impairs focal adhesion formation only under restricted conditions."J.Biol.Chem.. 275. 5249-5252 (2000)

[Publications] Yasushi Kobayashi: "Chaperones, Hsp70 and Hsp40, suppress aggregate formation and apoptosis in cultured neuronal cells expressing truncated androgen receptor protein with expanded polyglutamine."J.Biol.Chem.. (in press).

[Publications] Satoshi Fukumoto: "GD3 synthase gene expression in PC12 cells results in the continuous activation of TrkA and ERK1/2 and enhanced proliferation."J.Biol.Chem.. (in press).

[Publications] Mitsuru Horiba: "Neointima formation in a restenosis model is suppressed in midkine-deficient mice,"J.Clin.Invest.. (in press).

高橋雅英名古屋大学, 医学部, 教授 (40183446)