生体侵襲時のPerforinによる細胞膜障害機構の解明:生体内分子間相互作用解析装置を用いた解析

• 研究課題/領域番号: 14571221
• 研究種目: 基盤研究(C)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 消化器外科学
• 研究機関: 岩手医科大学
• 研究代表者: 佐藤 信博 岩手医科大学, 医学部, 講師 (10244914)
• 研究分担者: 前沢 千早 岩手医科大学, 医学部, 講師 (10326647)
• 研究期間 (年度): 2002 – 2003
• 研究課題ステータス: 完了 (2003年度)
• 配分額: 3,500千円 (直接経費: 3,500千円); 2003年度: 800千円 (直接経費: 800千円); 2002年度: 2,700千円 (直接経費: 2,700千円)
• キーワード: perforin / granzyme B / 生体内分子間相互作用 / maspin / センサーチップ / 生体内分子 / パーフォリン / CTL / リン脂質膜 / リ脂質膜

研究概要

生体への侵襲刺激によって,細胞内の傷害顆粒が分泌される.傷害顆粒からは,パーフォリンが放出され,細胞膜に細孔を形成する.この細孔を通過してグランザイムBが細胞内に導入され,標的細胞をアポトーシスに誘導する.パーフォリン/グランザイムBによる一連の細胞障害機構は,細胞障害機構として注目され,敗血症性ARDSの肺胞上皮や血管内皮細胞の主要な傷害因子であることが明らかとなった.パーフォリンによる細胞膜傷害機構の精巧なin vitroモデルを開発することは,生体に加わる様々な侵襲(感染・担癌・手術侵襲)に対する生体反応の病態診断に役立つばかりでなく,過剰な生体反応に対する治療薬の開発にも役立つ.当該研究課題では生体内分子間相互作用解析装置を用いて,リン脂質膜構造を金箔膜の張られたセンサーチップ上に形成し,パーフォリンによる細胞膜傷害活性の定量的評価システムを開発し,臨床応用の可能性を検討する.
本研究課題では研究期間内に,以下の成果を得た.
(1)パーフォリンによる細胞膜傷害活性の定量的評価システムを開発し,論文を投稿中である.
(2)現在までに,特定のパーフォリンの阻害作用をもつ薬剤の特定には至っていないが,serine protease inhibitor (serpin)を有効な標的分子として解析を行った.未だ,機能的には不明な分子であるが,mammary serine protease inhibitor (maspin)がその候補として考えられ,現在dominat negativeを作製し検討中である.
(3)granzyme Bの定量システムについては現在も引き続き検討中である.
パーフォリンの細胞障害機構は依然不明の点もあるが,癌抑制遺伝子maspinがその阻害作用を持つ可能性が示唆された,現在詳細を検討中であるが,RNAiを用いた,急性期の炎症性疾患治療戦略の可能性が示唆された.

研究成果

• [文献書誌] Sato N, Koeda K, Ikeda K, Kimura Y, Aoki K, Iwaya T, Akiyama Y, Ishida K, Saito K, Endo S.: "Randomized study of the benefits of preoperative corticosteroid administration on the postoperative morbidity and cytokine response in patients undergoing surgery."Ann Surg.. 236(2). 184-190 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Sato N, Endo S, Kimura Y, Ikeda K, Aoki K, Iwaya T, Akiyama Y, Noda Y, Saito K.: "Influence of a human protease inhibitor on surgical stress induced immunosuppression."Dig Surg.. 19(4). 300-305 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Maesawa C, Iijima S, Sato N, Yoshinori N, Suzuki M, Tarusawa M, Ishida K, Tamura G, Saito K, Masuda T.: "Esophageal extraskeletal Ewing's sarcoma."Hum Pathol.. 33(1). 130-132 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lymphoid malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction."Int J Hematol.. 75(2). 166-173 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Iijima S, Maesawa C, Sato N, Ikeda K, Inaba T, Akiyama Y, Ishida K, Saito K, Masuda T.: "Gastrointestinal stromal tumour of the oesophagus : significance of immunohistochemical and genetic analyses of the c-kit gene."Gastroenterol Hepatol.. 14(4). 445-448 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lymphoproliferative disorder with features of granulomatous slack skin disease."Br J Dermatol.. 147(5). 998-1002 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, Suzuki M, Yashima A, Ogasawara S, Oikawa H, Sato N, Saito K, Masuda T.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance."Nucleic Acids Res.. 31(2). E4-E4 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Yashima A, Maesawa C, Uchiyama M, Tarusawa M, Satoh T, Satoh M, Enomoto S, Sugawara K, Numaoka H, Murai K, Utsugisawa T, Ishida Y, Masuda T.: "Quantitative assessment of contaminating tumor cells in autologous peripheral blood stem cells of B-cell non-Hodgkin lymphomas using immunoglobulin heavy chain gene allele-specific oligonucleotide real-time quantitative-polymerase chain reaction."Leuk Res.. 27(10). 925-934 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Nakatani K, Hagihara S, Sando S, Sakamoto S, Yamaguchi K, Maesawa C, Saito I.: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer : inhibition of the elongation of a telomeric repeat by telomerase."J Am Chem Soc.. 125(3). 662-666 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Ishida Y, Masuda T.: "Consensus primers for detecting monoclonal immunoglobulin heavy chain rearrangement in B-cell lymphomas."J Clin Pathol.. 56(10). 778-779 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Satoh T, Ishida Y, Masuda T.: "Development of immunoglobulin variable heavy-chain gene consensus probes with conjugated 3'-minor groove binder groups for monitoring minimal residual disease in childhood acute lymphoblastic leukemia."J Clin Pathol.. 56(12). 952-955 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T.: "Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells."Am J Pathol.. 163(5). 1911-1919 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Tarusawa M, Satoh S, Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Motoyoshi K, Masuda T.: "Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas."Cancer Sci.. 94(10). 877-885 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ogasawara S, Maesawa C, Yamamoto M, Akiyama Y, Wada K, Fujisawa K, Higuchi T, Tomisawa Y, Sato N, Endo S, Saito K, Masuda T.: "Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers."Oncogene.. 23(5). 1117-1124 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Wada K, Maesawa C, Akasaka T, Masuda T.: "Aberrant expression of the maspin gene associated with epigenetic modification in melanoma cells."J Invest Dermatol.. 122(3). 805-811 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Oyama M, Terashima M, Takagane A, Maesawa C.: "Prognostic significance of peritoneal minimal residual disease in detecting mRNA in gastric cancer using quantitative real-time reverse transcriotive-polymerase chain reaction of carcinoembryonic antigen."Br J Surg.. 91(4). 435-443 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Itabashi T, Maesawa C, Uchiyama M, Higuchi T, Masuda T.: "Quantitative detection of mutant alleles of the K-ras gene with minor groove binder-conjugated fluorogenic DNA probes."Int J Oncol.. 24(3). 687-696 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Satoh H, Nakamura M, Satoh M, Nakajima T, Izumoto H, Maesawa C, Kawazoe K, Masuda T, Hiramori K.: "Expression and localization of Tumor Necrosis Factor-alpha and its converting enzyme in human abdominal aortic aneurysm."Clin Sci (Lond).. 106(3). 301-306 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Akiyama Y, Maesawa C, Wada K, Fujisawa K, Itabashi T, Noda Y, Honda T, Sato N, Ishida K, Takagane A, Saito K, Masuda T.: "Human PinX1, a potent telomerase inhibitor, is not involved in human gastrointestinal tract carcinoma."Oncol Rep.. 11(4). 871-874 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Sato N, Koeda K, Ikeda K, Kimura Y, Aoki K, Iwaya R, Akiyama Y, Ishida K, Saito K, Endo S.: "Randomized study of the benefits of preoperative corticosteroid administration on the postoperative morbidity and cytokine response in patients undergoing surgery for esophageal cancer."Ann Surg.. 235(2). 184-190 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Sato N, Endo S, Kimura Y, Ikeda K, Aoki K, Iwaya T, Akiyama Y, Noda Y, Saito K.: "Influence of a human protease inhibitor on surgical stress induced immunosuppression."Dig Surg. 19(4). 300-305 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Maesawa C, Iijina S, Sato N, Yoshinori N, Suzuki M, Tanisawa M, Ishida K, Tamura G, Saito K, Masuda T.: "Esophageal extraskeletal Ewing's sarcoma."Hum Pathol.. 33(1). 130-132 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lymphoid malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction."Int J Hematol.. 75(2). 166-173 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Iijima S, Maesawa C, Sato N, Ikeda K, Inaba T, Akiyama Y, Ishida K, Saito K, Masuda T.: "Gastrointestinal stromal tumour of the oesophagus: significance of immunohistochemical and genetic analyses of the c-kit gene."Eur J Gastroenterol Hepatol.. 14(4). 445-448 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lyimphoproliferative disorder with features of granulomatous slack skin disease."Br J Dermatol.. 147(5). 998-1002 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, Suzuki M, Yashima A, Ogasawara S, Oikawa H, Sato N, Saito K, Masuda T.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance."Nucleic Acids Res.. 31(2). E4-E4 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Yashima A, Maesawa C, Uchiyama M, Tarusawa M, Satoh T, Satoh M, Enomoto S, Sugawara K, Numaoka H, Murai K, Utsugisawa T, Ishida Y, Masuda T.: "Quantitative assessment of contaminating tumor cells in autologous peripheral blood stem cells of B-cell non-Hodgkin lymphomas using immunoglobulin heavy chain gene allele-specific oligonucleotide real-time quantitative-polymerase chain reaction."Leuk Res.. 27(10). 925-934 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Nakatani K, Hagihara S, Sando S, Sakamoto S, Yamaguchi K, Maesawa C, Saito I.: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer inhibition of the elongation of a telomeric repeat by telomerase."J Am Chem Soc.. 125(3). 662-666 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Ishida Y Masuda T.: "Consensus primers for detecting monoclonal immunoglobulin heavy chain rearrangement in B-cell lymphomas."J Chin Pathol.. 56(10). 778-779 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Satoh T, Ishida Y, Masuda T.: "Development of immunoglobulin variable heavy-chain gene consensus probes with conjugated 3'-minor groove binder groups for monitoring minimal residual disease in childhood acute lymphoblastic leukemia."J Chin Pathol.. 56(12). 952-955 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T.: "Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells."Am J Pathol.. 163(5). 1911-1919 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Tarusawa N, Satoh S< Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Motoyoshi K, Masuda T.: "Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas."Cancer Sci.. 94(10). 877-885 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ogasawara S, Maesawa C, Yamamoto M, Akiyama Y, Wada K, Fujisawa K, Higuchi T, Tomisawa Y, Sato N, Endo S, Saito K, Masuda T.: "Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers."Oncogene.. 23(5). 1117-1124 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Wada K, Maesawa C, Akasaka T, Masuda T.: "Aberrant expression of the maspin gene associated with epigenetic modification in melanoma cells."J Invest Dermatol.. 112(3). 805-811 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Oyama M, Terashima M, Takagane A, Maesawa C.: "Prognostic significance of peritoneal minimal residual disease in detecting mRNA in gastric cancer using quantitative real-time reverse transcriotive-polymerase chain reaction of carcinoembryonic antigen."Br J Surg.. 91(4). 435-443 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Itabashi T, Maesawa C, Uchiyama M, Higuchi T, Masuda T.: "Quantitative detection of mutant alleles of the K-ras gene with minor groove binder-conjugated fluorogenic DNA probes."Int J Oncol.. 24(3). 687-696 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Satoh H, Nakamura M, Satoh M, Nakajima T, Izumoto H, Maesawa C, Kawazoc K, Masuda T, Hiramori K.: "Expression and localization of Tumor Necrosis Factor-alpha and its converting enzyme in human abdominal aortic aneurysm."Clin Sci (Lond).. 106(3). 301-306 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Akiyama Y, Maesawa C, Wada K, Fujisawa K, Itabashi T, Noda Y, Honda T, Sato N, Ishida K, Takagane A, Saito K, Masuda T.: "Human PinX1, a potent telomerase inhibitor, is not involved in human gastrointestinal tract carcinoma."Oncol Re. 11(4). 871-874 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Akiyama Y, Maesawa C, Wada K, Fujisawa K, Itabashi T, Noda Y, Honda T, Sato N, Ishida K, Takagane A, Saito K, Masuda T: "Human PinX1, a potent telomerase inhibitor, is not involved in human gastrointestinal tract carcinoma."Oncol Rep. 11・4. 871-874 (2004)
• [文献書誌] Ogasawara S, Maesawa C, Yamamoto M, Akiyama Y, Wada K, Fujisawa K, Higuchi T, Tomisawa Y. Soto N, E: "Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers."Oncogene. 23・5. 1117-1124 (2004)
• [文献書誌] Yamada Y, Endo S, Nakae H, Makabe H, Sato N, Wakabayashi G, Kitamura M, Inada K, Sato S: "Examination of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients with burns."Burns. 29・8. 799-802 (2003)
• [文献書誌] Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T: "Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells."Am J Pathol. 165・5. 1911-1919 (2003)
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Tarusawa M.Satoh S, Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Motoyoshi K, Masuda T: "Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas."Cancer Sci. 94・10. 875-885 (2003)
• [文献書誌] Iijima S, Maesawa C, Sato N, et al.: "Gastrointestinal stromal tumour of the oesphagus : significance of immunohistochemical and genetic analyses of the c-kit gene"Eur J Gastroterol Hepatol. 14. 445-448 (2002)
• [文献書誌] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lymphoid malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction"Int J Hematol. 75. 166-173 (2002)
• [文献書誌] Sato N, Koeda K, Ikeda K, Kimura Y., et al.: "Randomized study of the benefit of preoperative corticosteroid administration on the postoperative morbidity and cytokine response in patients undergoing surgery for esophageal cancer"Ann Surg. 236. 184-190 (2002)
• [文献書誌] Nakatani K, Hagihara S, Sando S, Sakamoto S, Yamaguchi K, Maesawa C, Saito I.: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer : inhibition of the elongation of a telomeric repeat by telomerase"J Am Chem Soc. 125. 662-666 (2003)
• [文献書誌] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, et al.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance"Nucleic Acids Res. 31. E4-4 (2003)
• [文献書誌] Sato N, Endo S, Kimura Y, Ikeda K, et al.: "Influence of a human protease inhibitor on surgical stress induced immunosuppression"Dig Surg. 19. 300-305 (2002)