2002 年度実績報告書

疾患モデル動物の体系的遺伝子発現解析による創薬標的分子の同定

研究課題

研究課題/領域番号	14370039
研究機関	国立成育医療センター(研究所)
研究代表者	辻本豪三国立成育医療センター研究所, 薬剤治療研究部, 部長 (80172013)
研究分担者	輿水崇鏡国立成育医療センター研究所, 薬剤治療研究部, 研究員塩島聡国立成育医療センター研究所, 薬剤治療研究部, 研究員篠浦ひとみ国立成育医療センター研究所, 薬剤治療研究部, 研究員山田将輝国立成育医療センター研究所, 薬剤治療研究部, 研究員
キーワード	機能ゲノム科学 / マイクロアレイ / cDNAチップ / 疾患モデル動物 / 標準化cDNAライブラリー / 病態遺伝子
研究概要	ゲノムテクノロジーの進展により、微量検体で発現変動している遺伝子群を解析する技術革新としてマイクロアレイ法やDNAチップが登場した。特に、その経済性、網羅性の点からマイクロアレイ法は現在飛躍的にその普及性をのばしており、その応用はヒト及びマウス等のゲノムプロジェクトより得られつつある遺伝子情報と相まって、今後遺伝子機能解析の最有力な技術となることが期待されている。一方、薬理学において疾患モデル動物として薬効評価に従来より用いられてきている動物モデルはラット、モルモット、ウサギが主であり、その遺伝子解析はヒト、マウスと比して著しく遅れており、遺伝子情報に乏しい。このような背景の疾患動物モデルにおける体系的かつ網羅的な遺伝子発現解析が可能となれば、従来より蓄積されているこれら疾患モデル動物を用いた膨大な生理、生化学薬理データを遺伝子変動と関連して有効に活用でき、そのもたらす効果は計り知れないものがある。このような背景のもと、本研究ではその遺伝子情報が未だ充実していない動物モデル(ラット、モルモット)の各臓器別標準化cDNAライブラリーを作成し、このライブラリーを高密度に配置した臓器別標準cDNAライブラリーDNAチップを用い、数種のモデル疾患(腎炎、腎結石、肺繊維症、高血圧、糖尿病など)における病態遺伝子発現の解析を目的とした。本年度はラット腎臓標準化cDNAマイクロアレイチップを用いてラット糸球体腎炎モデル、腎結石モデル等を対象として解析し、腎疾患の病態形成,病状の悪や治療のターゲットに関連する可能性の極めて高い遺伝子群を見出すことができた。本研究により作製したラット腎臓標準化cDNAマイクロアレイチップは、腎疾患の病状の悪化、あるいは改善に関する遺伝子を見出すために有用であると考えられる。疾患に関与する遺伝子が明らかになれば、それをターゲットとしたゲノム創薬がスタートできることから、本研究の意義は大きい。

研究成果

(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Sugawara T, Hirasawa A, Hashimoto K, Tsujimoto G.: "Differences in the subcellular localization of α 1-adrecoceptor subtypes can affect the subtype selectivity of drugs in a study with the fluorescent ligand BODIPY-FL prazosin"Life Sciences.. 70. 2113-2124 (2002)
[文献書誌] Sugimoto Y, Tsujimoto G, Nagao T, Kurose H, et al.: "β1-Selective agonist (-)-1-(3,4-Dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanal [(-)-RO363] differentially interacts with key amino acids responsible for β1-selective binding in resting and active states"J. Pharamacol. Exp. Ther.. 301. 51-58 (2002)
[文献書誌] Chalothorn D, McCune D.F, Tsujimoto G, Piascik M.T., et al.: "Differences in the cellular localization and agonist mediated internalizaion properties of the alpha1-adrenoceptor subtypes"Mol. Pharmacol.. 61(5). 1008-1016 (2002)
[文献書誌] Koshimizu T, Yamauchi J, Hirasawa A, Tanoue A, Tsujimoto G.: "Recent progress in α_1-adrenoceptor pharmacology"Biol. Pharm. Bull.. 25. 401-408 (2002)
[文献書誌] Tanoue A, Nasa Y, Shinoura H, Takeo S, Tsujimoto G, et al.: "The α_<1D>-adrenergic receptor directly regulates arterial blood pressure via vasoconstriction"J Clin. Invest.. 109. 765-775 (2002)
[文献書誌] Tanoue A, Koshimizu T, Tsuchiya M, Ishii K, Tsujimoto G, et al.: "Two novel transcripts for human endothelin B receptor produced by RNA editing/alternativesplicing from a single gene"J. Biol. Chem.. 277. 33205-33212 (2002)
[文献書誌] Tanoue A, Koshimizu T, Hosoda C, Oshikawa S, Tsujimoto G. et al.: "Role of the α1D-adrenergic receptor in the development of salt-induced hypertension"Hypertension. 40. 101-106 (2002)
[文献書誌] Yamauchi J, Hirasawa A, Sugawara Y, Tsujimoto G, Itoh H, et al.: "Role of Dbl's big sister in the anti-mitogenic pathway from alpha1B-"Biochem. Biophys. Res. Commun.. 296. 85-92 (2002)
[文献書誌] Katsuma S, Shiojima S, Hirasawa A, Yano J, Tsujimoto G, et al.: "Global analysis of differentially expressed genes during progression of calcium oxalate nephrolithiasis"Biochem Biophys Res Commun.. 296. 544-551 (2002)
[文献書誌] Yoshida N, Nishimaki Y, Tanoue A, Hirasawa A, Tsujimoto G, et al.: "SNP genotyping in the β2-adrenergic receptor by electronic microchip assay, DHPLC, and direct sequencing"J. Hum. Genet.. 47. 500-503 (2002)
[文献書誌] Yamauchi J, Miyamoto Y, Hirasawa A, Tsujimoto G, Itoh H, et al.: "Endothelin suppresses cell migration via the JNK signaling pathway in a manner dependent upon Src kinase, Rac1, and Cdc42"FEBS Lett.. 527(1-3). 284-288 (2002)
[文献書誌] Shinoura H, Tsujimoto G, Teranishi Y, Tsuru H.: "Antagonistic effects of antimuscarinic drugs on α1-adrenoceptors"Naunyn Schmiedebergs Arch Pharmacol.. 366(4). 368-371 (2002)
[文献書誌] Hashizume K, S.Katsuma S, Shiojima S, Hirasawa A, Tsujimoto G, Izaike Y, et al.: "Implantation and placental development in somatic cell clone recipient cows"Cloning and Stem Cells.. 4(3). 197-209 (2002)
[文献書誌] Heishi M, Kagaya S, Sugita Y, Tsujimoto G, Saito H, et al.: "High-density oligonucleotide array analysis of mRNA transcripts in peripheral blood cells of severe atopic dermatitis patients"Int. Arch. Allergy Immunol.. 129. 57-66 (2002)
[文献書誌] Takei Y, Assenberg M, Tsujimoto G, Laskey R.: "The MCM3 acetylase MCM3AP inhibits initiation, but not elongation of DNA replication via interaction with MCM3"J. Biol. Chem.. 277. 43121-43125 (2002)
[文献書誌] Nakajima T, Ebisawa M, Tsujimoto G, Matsuda H, Nagai H, Saito H, et al.: "Marked increase in CC chemokine gene expression in both human and mouse mast cell transcriptomes folllowing Fce resecptor I cross-linking"Blood. 100. 3861-3868 (2002)
[文献書誌] Imai Y, Tsujimoto G.Matsumoto K, Akasawa A, Saito H, Oshida T, et al.: "Cloning and characterization of the highly expressed ETEA gene from blood cells of atopic dermatitis patients"Biochem. Biophys. Res. Commun.. 297. 1282-1290 (2002)
[文献書誌] Shinoura H, Shibata K, Hirasawa A, Tanoue A, Hashimoto K, Tsujimoto G.: "Key amino acids for differential coupling of α1-adrenergic receptor subtypes to Gs"Biochem. Biophys. Res. Commun.. 299. 142-147 (2002)
[文献書誌] Koshimizu TA, Ueno S, Tanoue A, Yanagihara N, Stanko SS, Tsujimoto G: "Heteromultimerization modulates P2X receptor-functions through participating extracellular and C-terminal subdomains"J Biol Chem.. 277(49). 46891-46899 (2002)
[文献書誌] Matsumoto Y, Oshida T, Tanaka T, Taujimoto G, Hirasawa A, Saito H, et al.: "Identification of highly expressed genes in peripheral blood T cells from patients with atopic dermatitis"Int Arch Allergy Immunol.. 129(4). 327-340 (2002)
[文献書誌] Katsuma S, Tanoue A, Takagaki K, Ohgi T, Yano J, Tsujimoto G, et al.: "Signaling mechanisms in sphingosine 1 -phosphate-promoted mesangial cell proliferation"Genes to Cells.. 7(12). 1217-1230 (2002)
[文献書誌] Shibata K, Katsuma S, Koshimizu T, Shinoura H, Hirasawa A, Tsujimoto G.: "Alpha 1-Adrenergic receptor subtypes differentially control cell cycle of transfected CHO cells through a cAMP-dependent mechanism involving p27Kip 1"J Biol Chem.. 278(1). 672-678 (2003)
[文献書誌] Katsuma S, Tanoue A, Takagaki K, Ohgi T, Yano J, Tsujimoto G. et al.: "Transcriptional profiling of gene expression patterns during sphingosine 1-phosphate-induced mesangial cell proliferation"Biochem. Biophys. Res. Commun.. 300. 577-584 (2003)
[文献書誌] Ishiwata H, Katsuma S, Hirasawa A, Tsujimoto G, Hashizume K, et al.: "Characterization of gene expression profiles in early bovine pregnancy using a custom cDNA microarray"Mol Reprod Dev.. 65. 9-18 (2003)
[文献書誌] Yoshida N, Sugiyama M, Tanoue A, Hirasawa A. Tsujimoto G.: "Determination of a novel haplotype of β2-adrenergic receptor in the Japanese population by the combination of the electronic microchip assay using the Nano Chip system with allele-specific PCR"Drug Metabolism and Pharmacokinetics. 17(6). 532-539 (2003)
[文献書誌] Koshimizu TA, Tanoue A, Hirasawa A, Yamauchi J, Tsujimoto G.: "Recent advances in al-adrenoceptor pharmacology"Pharmacology & Therapeutics. 5535(in press). 1-10 (2003)
[文献書誌] Nakamura T, Shiojima S, Iwama T, Katsuma S, Tsujimoto G, et al.: "Temporal gene expression changes of adipogenesis in human mesenchymal stem cells"Biochem. Biophys. Res. Commun.. 303. 306-312 (2003)

2002 年度 実績報告書

疾患モデル動物の体系的遺伝子発現解析による創薬標的分子の同定

研究代表者

辻本 豪三 国立成育医療センター研究所, 薬剤治療研究部, 部長 (80172013)

研究成果

[文献書誌] Sugawara T, Hirasawa A, Hashimoto K, Tsujimoto G.: "Differences in the subcellular localization of α 1-adrecoceptor subtypes can affect the subtype selectivity of drugs in a study with the fluorescent ligand BODIPY-FL prazosin"Life Sciences.. 70. 2113-2124 (2002)

[文献書誌] Chalothorn D, McCune D.F, Tsujimoto G, Piascik M.T., et al.: "Differences in the cellular localization and agonist mediated internalizaion properties of the alpha1-adrenoceptor subtypes"Mol. Pharmacol.. 61(5). 1008-1016 (2002)

[文献書誌] Koshimizu T, Yamauchi J, Hirasawa A, Tanoue A, Tsujimoto G.: "Recent progress in α_1-adrenoceptor pharmacology"Biol. Pharm. Bull.. 25. 401-408 (2002)

[文献書誌] Tanoue A, Nasa Y, Shinoura H, Takeo S, Tsujimoto G, et al.: "The α_<1D>-adrenergic receptor directly regulates arterial blood pressure via vasoconstriction"J Clin. Invest.. 109. 765-775 (2002)

[文献書誌] Tanoue A, Koshimizu T, Tsuchiya M, Ishii K, Tsujimoto G, et al.: "Two novel transcripts for human endothelin B receptor produced by RNA editing/alternativesplicing from a single gene"J. Biol. Chem.. 277. 33205-33212 (2002)

[文献書誌] Tanoue A, Koshimizu T, Hosoda C, Oshikawa S, Tsujimoto G. et al.: "Role of the α1D-adrenergic receptor in the development of salt-induced hypertension"Hypertension. 40. 101-106 (2002)

[文献書誌] Yamauchi J, Hirasawa A, Sugawara Y, Tsujimoto G, Itoh H, et al.: "Role of Dbl's big sister in the anti-mitogenic pathway from alpha1B-"Biochem. Biophys. Res. Commun.. 296. 85-92 (2002)

[文献書誌] Katsuma S, Shiojima S, Hirasawa A, Yano J, Tsujimoto G, et al.: "Global analysis of differentially expressed genes during progression of calcium oxalate nephrolithiasis"Biochem Biophys Res Commun.. 296. 544-551 (2002)

[文献書誌] Yoshida N, Nishimaki Y, Tanoue A, Hirasawa A, Tsujimoto G, et al.: "SNP genotyping in the β2-adrenergic receptor by electronic microchip assay, DHPLC, and direct sequencing"J. Hum. Genet.. 47. 500-503 (2002)

[文献書誌] Yamauchi J, Miyamoto Y, Hirasawa A, Tsujimoto G, Itoh H, et al.: "Endothelin suppresses cell migration via the JNK signaling pathway in a manner dependent upon Src kinase, Rac1, and Cdc42"FEBS Lett.. 527(1-3). 284-288 (2002)

[文献書誌] Shinoura H, Tsujimoto G, Teranishi Y, Tsuru H.: "Antagonistic effects of antimuscarinic drugs on α1-adrenoceptors"Naunyn Schmiedebergs Arch Pharmacol.. 366(4). 368-371 (2002)

[文献書誌] Hashizume K, S.Katsuma S, Shiojima S, Hirasawa A, Tsujimoto G, Izaike Y, et al.: "Implantation and placental development in somatic cell clone recipient cows"Cloning and Stem Cells.. 4(3). 197-209 (2002)

[文献書誌] Heishi M, Kagaya S, Sugita Y, Tsujimoto G, Saito H, et al.: "High-density oligonucleotide array analysis of mRNA transcripts in peripheral blood cells of severe atopic dermatitis patients"Int. Arch. Allergy Immunol.. 129. 57-66 (2002)

[文献書誌] Takei Y, Assenberg M, Tsujimoto G, Laskey R.: "The MCM3 acetylase MCM3AP inhibits initiation, but not elongation of DNA replication via interaction with MCM3"J. Biol. Chem.. 277. 43121-43125 (2002)

[文献書誌] Nakajima T, Ebisawa M, Tsujimoto G, Matsuda H, Nagai H, Saito H, et al.: "Marked increase in CC chemokine gene expression in both human and mouse mast cell transcriptomes folllowing Fce resecptor I cross-linking"Blood. 100. 3861-3868 (2002)

[文献書誌] Imai Y, Tsujimoto G.Matsumoto K, Akasawa A, Saito H, Oshida T, et al.: "Cloning and characterization of the highly expressed ETEA gene from blood cells of atopic dermatitis patients"Biochem. Biophys. Res. Commun.. 297. 1282-1290 (2002)

[文献書誌] Shinoura H, Shibata K, Hirasawa A, Tanoue A, Hashimoto K, Tsujimoto G.: "Key amino acids for differential coupling of α1-adrenergic receptor subtypes to Gs"Biochem. Biophys. Res. Commun.. 299. 142-147 (2002)

[文献書誌] Koshimizu TA, Ueno S, Tanoue A, Yanagihara N, Stanko SS, Tsujimoto G: "Heteromultimerization modulates P2X receptor-functions through participating extracellular and C-terminal subdomains"J Biol Chem.. 277(49). 46891-46899 (2002)

[文献書誌] Matsumoto Y, Oshida T, Tanaka T, Taujimoto G, Hirasawa A, Saito H, et al.: "Identification of highly expressed genes in peripheral blood T cells from patients with atopic dermatitis"Int Arch Allergy Immunol.. 129(4). 327-340 (2002)

[文献書誌] Katsuma S, Tanoue A, Takagaki K, Ohgi T, Yano J, Tsujimoto G, et al.: "Signaling mechanisms in sphingosine 1 -phosphate-promoted mesangial cell proliferation"Genes to Cells.. 7(12). 1217-1230 (2002)

[文献書誌] Shibata K, Katsuma S, Koshimizu T, Shinoura H, Hirasawa A, Tsujimoto G.: "Alpha 1-Adrenergic receptor subtypes differentially control cell cycle of transfected CHO cells through a cAMP-dependent mechanism involving p27Kip 1"J Biol Chem.. 278(1). 672-678 (2003)

[文献書誌] Katsuma S, Tanoue A, Takagaki K, Ohgi T, Yano J, Tsujimoto G. et al.: "Transcriptional profiling of gene expression patterns during sphingosine 1-phosphate-induced mesangial cell proliferation"Biochem. Biophys. Res. Commun.. 300. 577-584 (2003)

[文献書誌] Ishiwata H, Katsuma S, Hirasawa A, Tsujimoto G, Hashizume K, et al.: "Characterization of gene expression profiles in early bovine pregnancy using a custom cDNA microarray"Mol Reprod Dev.. 65. 9-18 (2003)

[文献書誌] Koshimizu TA, Tanoue A, Hirasawa A, Yamauchi J, Tsujimoto G.: "Recent advances in al-adrenoceptor pharmacology"Pharmacology & Therapeutics. 5535(in press). 1-10 (2003)

[文献書誌] Nakamura T, Shiojima S, Iwama T, Katsuma S, Tsujimoto G, et al.: "Temporal gene expression changes of adipogenesis in human mesenchymal stem cells"Biochem. Biophys. Res. Commun.. 303. 306-312 (2003)

2002 年度実績報告書

辻本豪三国立成育医療センター研究所, 薬剤治療研究部, 部長 (80172013)