メサンギウム細胞特異発現遺伝子群の機能解析及びその病因論に関する研究

• 研究課題/領域番号: 11307016
• 研究種目: 基盤研究(A)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 腎臓内科学
• 研究機関: 東海大学
• 研究代表者: 黒川 清 東海大学, 医学部, 教授 (30167390)
• 研究分担者: 稲城 玲子 東海大学, 総合医学研究所, 講師 (50232509) ; 宮田 敏男 東海大学, 医学部, 助教授 (10222332)
• 研究期間 (年度): 1999 – 2000
• 研究課題ステータス: 完了 (2000年度)
• 配分額: 38,200千円 (直接経費: 38,200千円); 2000年度: 17,500千円 (直接経費: 17,500千円); 1999年度: 20,700千円 (直接経費: 20,700千円)
• キーワード: 3'-directed cDNA library法 / メサンギウム細胞 / スサンギウム細胞特異的遺伝子 / megsin / 進行性糸球体疾患 / 分子生物学 / メサンギウム細胞特異的遺伝子 / 3'-directed / cDNA / library法

研究概要

従来、細胞特異的遺伝子の同定にはsubtraction library法やdifferential hybridization法などが用いられてきたが、いずれも定量性に欠き、手技的な条件設定の困難さが残されている。そこで我々は、ヒトメサンギウム細胞について、より定量的で、なおかつ細胞特異的遺伝子を検出しやすいcDNAライブラリーの作製を考慮し、近年確立された3'-directed cDNA library法を起用した。
この独創的な分子生物学的アプローチによって、1)世界に先駆け、ヒトメサンギウム細胞に発現する遺伝子群の定量的解析を行い、細胞特性の一部を分子レベルで明らかにすることに成功し、その中から5つの未知のメサンギウム細胞特異的遺伝子(megsin,meg-1,-2,-3,-4と命名)の単離同定を行った。2)megsinはメサンギウム細胞に高発現し、既知のセリンプロテアーゼインヒビター(serpin)と高い相同性を有する未知の蛋白をコードする新規遺伝子で、メサンギウム増殖性IgA腎症でその発現が亢進する。3)meg-1,meg-2,meg-3,meg-4もmegsinと同様、ヒトのみならずラット、マウスなど種を越えてメサンギウム細胞に保存され、なおかつ高発現している新規遺伝子で、いずれも未知の蛋白を一つコードする。アミノ酸相同性や蛋白モチーフの検索からmeg-1はserine/threonin alkarine phosphatase4の調節ユニット,meg-3は高プロリンドメインを有する転写調節因子、meg-4はATP依存性metalloproteaseである可能性が示唆された。

研究成果

• [文献書誌] Suzuki D et. al: "Immunohistochemical evidence for an increased oxidative stress and carbonylmodification of proteins in diabetic glomerular lesions."J Am Soc Nephrol. 10(4). 822-832 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Miyata T et. al: "Apolipoprotein E2/E5 variants in lipoprotein glomerulopathy recurred in transplanted kidney."J Am Soc Nephrol. 10(7). 1590-1595 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Kawada N et. al: "Increased oxidative stress in mouse kidneys with unilateral ureteral obstruction."Kidney Int. 56(3). 1004-1013 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Suzuki D et. al: "Expression of megsin mRNA,a novel mesangium-predominant gene,in the renaltissues of various glomerular diseases."J Am Soc Nephrol. 10(12). 2606-2613 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Suzuki D, Miyata T, Saotome N, Horie K, Inagi R, Yasuda Y, Uchida K, Izuhara Y, Yagame M, Sakai H, Kurokawa K: "Immunohistochemical evidence for an increased oxidative stress and carbonyl modification of proteins in diabetic glomerular lesions."J Am Soc Nephrol. 10 (4). 822-832 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Miyata T, Sugiyama S, Nangaku M, Suzuki D, Uragami K, Inagi R, Kurokawa K: "Apolipoprotein E2/E5 variants in lipoprotein glomerulopathy recurred in transplanted kidney"J Am Soc Nephrol. 10 (7). 1590-1595 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Kawada N, Moriyama T, Ando A, Fukunaga M, Miyata T, Kurokawa K, Imai E, Hori M.: "Increased oxidative stress in mouse kidneys with unilateral ureteral obstruction."Kidney Int. 56 (3). 1004-1013 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Suzuki D, Miyata T, Nangaku M, Takano H, Saotome N, Toyoda M, Mori Y, Zhang S, Inagi R, Endoh M, Kurokawa K, Sakai H.: "Expression of megsin mRNA, a novel mesangium-predominant gene, in the renal tissues of various glomerular diseases"J Am Soc Nephrol. 10 (12). 2606-2613 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
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• [文献書誌] Jadoul M et. al: "Influence of hemodialysis membrane type on pentosidine plasma level, a marker of "carbonyl stress""Kidney Int. 55. 2487-2492 (1999)
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