Development of inflammatory cell-specific gene regulation system for gene therapeutic strategy

• Project/Area Number: 20790042
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: St.Marianna University School of Medicine
• Principal Investigator: ASAI Daisuke St.Marianna University School of Medicine, 医学部, 助教 (10423485)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2009: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2008: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: IkappaBkinase / NF-kappaB / 遺伝子キャリアー / ドラッグデリバリー / 遺伝子治療 / IkappaB kinase

Research Abstract

We proposed a novel strategy called D-RECS (drug or gene delivery system responding to cellular signals) to convert an intracellular signal to transgene expression. Here we apply this concept to inflammatory cells by using Ix-B kinase as a signal molecule that triggers the gene expression. Candidate cationic substrates of Ix-B kinase beta (IKK.(3) were synthesized and their reactivity was investigated. The results of gel shift assay showed that the polyplex was disintegrated and free DNA was released in the presence of IKKf3. The polyplex comprising CMV-GFP plasmid DNA and the polymer expressed the transgene in living cells exposed to a pro-inflammatory stimulus. Our concept of cell-specific gene expression was proved to work in inflammatory cells. This method may provide a unique strategy for gene therapy exclusively in inflammatory cells.

Research Products

• [Journal Article] Specific transgene expression in HIV-infected cells using protease-cleavable transcription regulator. (2010)
  • Author(s): Asai, D., Kuramoto, M., Shoji, Y., Kang, J.H., Kodama, B.K., Kawamura, K., Mori, T., Miyoshi, H., Niidome, T., Nakashima, H., Katayama, Y.
  • Journal Title: J Control. Release 141 Pages: 52-61
  • Peer Reviewed
• [Journal Article] Plasma protein kinase C (PKC)αas a biomarker for the diagnosis of cancers. (2009)
  • Author(s): Kang, J.H. *, Asai, D. *, Toita, R., Kitazaki, H., Katayama, Y. * (*: ontributed equally)
  • Journal Title: Carcinogenesis 30 Pages: 1927-1931
  • Peer Reviewed
• [Journal Article] Inflammatory cell-specific gene regulation system responding to Iκ-B kinase beta activation. (2009)
  • Author(s): Asai, D., Tsuchiya, A., Kang, J.H., Kawamura, K., Oishi, J., Mori, T., Niidome, T., Shoji, Y., Nakashima, H., Katayama, Y.
  • Journal Title: J. Gene Med. 11 Pages: 624-632
  • Peer Reviewed
• [Journal Article] Regulation of transgene expression in tumor cells by exploiting endogenous intracellular signals. (2009)
  • Author(s): Asai, D., Kang, J.H., Toita, R., Tsuchiya, A., Niidome, T., Nakashima, H., Katayama, Y.
  • Journal Title: Nanoscale Res. Lett. 4 Pages: 229-233
  • Peer Reviewed
• [Journal Article] Cellular signal-specific peptide substrate is essential for the gene delivery system responding to cellular signals. (2009)
  • Author(s): Kang, J.H., Toita, R., Tomiyama, T., Oishi, J., Asai, D., Mori, T., Niidome, T., Katayama, Y.
  • Journal Title: Bioorg. Med. Chem. Lett. 19 Pages: 6082-6086
  • Peer Reviewed
• [Journal Article] Molecular mechanism of caspase-3-induced gene expression of polyplexes formed from polycations grafted with cationic substrate peptides. (2009)
  • Author(s): Kawamura, K., Kuramoto, M., Mori, T., Toita, R., Oishi, J., Sato, Y., Kang, J.H., Asai, D., Niidome, T., Katayama, Y.
  • Journal Title: J. Biomater. Sci. Polym. Ed. 20 Pages: 967-980
  • Peer Reviewed
• [Journal Article] Inflaminatory cell-specific gene regulation system responding to Iκ-Bkinase beta activation (2009)
  • Author(s): Asai, D., et al.
  • Journal Title: J.Gene Med. Pages: 624-631
  • Peer Reviewed
• [Journal Article] Specific transgene expression in HIV-infected cells using protease-cleavable transcription regulator (2009)
  • Author(s): Asai, D., et al.
  • Journal Title: J.Control. Release (印刷中)
  • Peer Reviewed
• [Journal Article] Plasma protein kinase C(PKC)α as a biomarker for the diagnosis of cancers (2009)
  • Author(s): Kang, J.H., et al.
  • Journal Title: Carcinogenesis (印刷中)
  • Peer Reviewed
• [Journal Article] Design of polymeric carriers for cancer-specific gene targeting: Utilization of abnormal protein kinase Ca activation in cancer cells. (2008)
  • Author(s): Kang, J.H., Asai, D., Kim, J.H., Mori, T., Toita, R., Tomiyama, T., Asami, Y., Oishi, J., Sato, Y., Niidome, T., Jun, B., Nakashima, H., Katayama, Y.
  • Journal Title: J. Am. Chem. Soc. 130 Pages: 14906-14907
  • Peer Reviewed
• [Journal Article] Direct measure of fluorescence intensity for efficient receptor-binding assay:Conjugates of ethinylcarboxyestradiol and 5(and 6)-carboxyfluorescein via a, w-diaminoalkanes as a tracer for estrogen receptor. (2008)
  • Author(s): Asai, D., Tokunaga, T., Kondo, K., Kawaguchi, T., Takayanagi, S., Shinmyozu, T., Nakai, M., Yakabe, Y., Shimohigashi, Y.
  • Journal Title: J. Biochem. 143 Pages: 781-792
  • Peer Reviewed
• [Journal Article] A short peptide is a protein kinase C (PKC) a-specific substrate. (2008)
  • Author(s): Kang, J.H. *, Asai, D. *, Yamada, S. *, Toita, R., Oishi, J., Mori, T., Niidome, T., Katayama, Y. (*: contributed equally)
  • Journal Title: Proteomics 8 Pages: 2006-2011
  • Peer Reviewed
• [Journal Article] Drug delivery system based on responses to an HIV infectious signal. (2008)
  • Author(s): Asai, D., Kodama, B.K., Shoji, Y., Nakashima, H., Kawamura, K., Oishi, J., Kuramoto, M., Niidome, T., Katayama, Y.
  • Journal Title: Med. Chem. 4 Pages: 386-391
  • Peer Reviewed
• [Journal Article] Disordered cell-specific gene regulation system for suicide gene therapy. (2008)
  • Author(s): Asai, D., Nakashima, H.
  • Journal Title: Jpn. J. Chemother 56 Pages: 538-542
  • NAID: 10022594885
  • Peer Reviewed
• [Journal Article] Correlation between phosphorylation ratios by MALDI-TOF MS analysis and enzyme kinetics. (2008)
  • Author(s): Kang, J.H., Kuramoto, M., Tsuchiya, A., Toita, R., Asai, D., Sato, T.Y., Mori, T., Niidome, T., Katayama, Y.
  • Journal Title: Eur J. Mass Spectrom. 14 Pages: 261-265
  • Peer Reviewed
• [Journal Article] Design of Polymeric Carriers for Cancer- specific Gene Targeting : Util ization of Abnormal Activation of Protein Kinase Cα in Cancer Cells (2008)
  • Author(s): Kang JH, et.al.
  • Journal Title: J. Am. Chem. Soc 130 Pages: 14906-14907
  • Peer Reviewed
• [Journal Article] Regulation of Transgene Expression in Tumor Cells by Exploiting Int racellular Signals (2008)
  • Author(s): Asai D, et.al.
  • Journal Title: Nanoscale Res. Lett 4 Pages: 229-233
  • Peer Reviewed
• [Journal Article] Disordered Cell-specific Gene Regulation System for Suicide Gene Therapy (2008)
  • Author(s): Asai D, et.al.
  • Journal Title: Jpn. J. Chemother 56 Pages: 538-542
  • NAID: 10022594885
  • Peer Reviewed
• [Journal Article] Correlation Between Phosphorylation Ratios by MALDI-TOF MS Analysis and Enzyme Kinetics (2008)
  • Author(s): Kang JH, et.al.
  • Journal Title: Eur J. Mass Spectrom 14 Pages: 261-265
  • Peer Reviewed
• [Journal Article] Hepatoma-targeted gene delivery using a tumor cell-specific gene regulation system combined with a human liver cell-specific bionanocapsule.
  • Author(s): Kang, J.H., Oishi, J., Kim, J.H., Ijuin, M., Toita, R., Jun, B., Asai, D., Mori, T., Niidome, T., Tanizawa, K., Kuroda, S., Katayama, Y.
  • Journal Title: Nanomedicine (in press)
  • Peer Reviewed
• [Journal Article] Progress in current non-viral carriers for gene delivery.
  • Author(s): Shoji-Moskowitz, Y, Asai, D., Kodama, K., Katayama, Y., Nakashima, H.
  • Journal Title: Frontiers in Medicinal Chemistry (in press)
• [Journal Article] Molecular Mechanism of Caspase- 3- induced Gene Expression of Poly plexes Formed from Polycations Grafted with Cationic Substrate Peptides
  • Author(s): Kawamura K, et.al.
  • Journal Title: J. Biomater. Sci. Polym. Ed (印刷中)
  • Peer Reviewed
• [Presentation] 疾患細胞特異的な遺伝子発現制御法の開発-第2報 (2009)
  • Author(s): 浅井大輔、姜貞勲、三好洋、片山佳樹、中島秀喜
  • Organizer: 第65回神奈川県感染症医学会学術集会
  • Place of Presentation: 横浜
  • Year and Date: 2009-03-21
• [Presentation] 疾患細胞特異的な遺伝子発現制御法の開発-第2報 (2009)
  • Author(s): 浅井大輔, ほか
  • Organizer: 第65回神奈川県感染症医学会
  • Place of Presentation: 横浜
  • Year and Date: 2009-03-21
• [Presentation] Inflammatory signal-sensing polymer responding to IK-B kinase beta act ivat ion. (2009)
  • Author(s): Asai, D., Tsuchiya, A., Kang, J.H., Niidome, T., Nakashima, H., Ka tayama, Y.
  • Organizer: 8th International Symposium on Front iers in Biomedical Polymers (FBPS2009)
  • Place of Presentation: Mishima
• [Presentation] Specific transgene expression in virus-infected cells by a virus protease-cleavable transcription suppressor. (2009)
  • Author(s): Asai, D., Kuramo to, M., Kang, J.H., Niidome, T., Miyoshi, H., Nakashima, H., Katayama, Y.
  • Organizer: BIT Life Sciences' 2nd Annual Protein and Peptide Conference (PepCon-2009)
  • Place of Presentation: Seoul, South Korea
• [Presentation] Specific transgene expression in virus-infected cells by a virus protease-cleavable transcription suppressor (2009)
  • Author(s): Asai, D., et al.
  • Organizer: BIT Life Sciences' 2^<nd> Annual Protein and Peptide Conference (PepCon-2009)
  • Place of Presentation: South Korea
• [Presentation] Inflammatory signal-sensing polymer responding to Iκ-B kinase bet a achvation (2009)
  • Author(s): Asai, D., et al.
  • Organizer: 8^<th> International Symposium on Frontiers in Biomedical Polyxners (FBPS2009)
  • Place of Presentation: Mishima
• [Presentation] PKCα-responsive gene regulator for cancer cell-specific imaging and therapy (2009)
  • Author(s): Katayama, Y., et al.
  • Organizer: 8^<th> International Symposium on Frontiers in Biomedical Polyxners (FBPS2009)
  • Place of Presentation: Mishima
• [Presentation] HIVプロテアーゼ阻害能を示した低分子化合物 (2009)
  • Author(s): 金本大成, ほか
  • Organizer: 第57回日本化学療法学会総会
  • Place of Presentation: Tokyo
• [Presentation] Granulicatella elegans由来のアルギニン水解酵素によるリンパ球増殖抑制 (2008)
  • Author(s): 金本大成, ほか
  • Organizer: 第64回神奈川感染症医学会
  • Place of Presentation: 横浜
  • Year and Date: 2008-09-27
• [Presentation] 癌細胞特異的シグナル応答型遺伝子送達システム (2008)
  • Author(s): 姜貞勲, ほか
  • Organizer: 遺伝子・デリバリー研究会第8回シンポジウム
  • Place of Presentation: 大阪
  • Year and Date: 2008-05-09
• [Presentation] 抗ウィルス薬開発の新戦略-ウイルス感染細胞特異的な遺伝子発現制御法の創出 (2008)
  • Author(s): 浅井大輔、三好洋、中島秀喜
  • Organizer: 第56回日本ウイルス学会学術集会
  • Place of Presentation: 岡山
• [Presentation] Tumor intracellular signal-targeting by application of specific substrate peptide for protein kinasecc (2008)
  • Author(s): Asai, D., Knag, J.H., Toita, R., Tomiyama, T., Mori, T., Niidome, T., Nakashima, H., Katayama, Y.
  • Organizer: 17' Meeting of Methods in Protein Structure Analysis
  • Place of Presentation: Sapporo
• [Presentation] HIV感染シグナルに応答する遺伝子発現制御材料の開発 (2008)
  • Author(s): 浅井大輔、倉本政則、森健、新留琢郎、東海林洋子、片山佳樹、中島秀喜
  • Organizer: 第24回日本DDS学会学術集会
  • Place of Presentation: 東京
• [Presentation] 細菌由来のアルギニン水解酵素によるリンパ球増殖抑制 (2008)
  • Author(s): 金本大成, ほか
  • Organizer: 第82回日本感染症学会総会
  • Place of Presentation: 松江
• [Presentation] 人工遺伝子転写制御システムを用いる細胞内シグナルイメージング (2008)
  • Author(s): 片山佳樹, ほか
  • Organizer: 日本ケミカルバイオロジー研究会第3回年会
  • Place of Presentation: 東京
• [Presentation] 癌細胞を標的とした細胞内シグナル応答型遺伝子送達システム (2008)
  • Author(s): 姜貞勲, ほか
  • Organizer: 第17回化学とマイクロ・ナノシステム研究会
  • Place of Presentation: 福岡
• [Presentation] 細胞内シグナルを利用する標的疾患細胞特異的遺伝子送達 (2008)
  • Author(s): 片山佳樹, ほか
  • Organizer: 日本薬剤学会第23回年会
  • Place of Presentation: 札幌
• [Presentation] ウイルス感染細胞選択的な遺伝子発現制御システムの構築 (2008)
  • Author(s): 倉本政則, ほか
  • Organizer: 第57回高分子学会
  • Place of Presentation: 横浜
• [Presentation] A novel gene carrier responding to virus protease (2008)
  • Author(s): Kuramoto, M., et.al.
  • Organizer: 35^<th> Annual Meeting & Expression of the Controlled Release Society
  • Place of Presentation: New York, USA
• [Presentation] Tumor intracellular signal- targeting by application of speci ficsubstrate peptide for protein kinase Cα (2008)
  • Author(s): Asai, D., et.al.
  • Organizer: 17^<th> Meeting of Methods in Protein Structure Analysis
  • Place of Presentation: Sapporo
• [Presentation] 新規クロマチン機能モデル-遺伝子転写を制御する高分子 (2008)
  • Author(s): 片山佳樹, ほか
  • Organizer: 第57回高分子討論会
  • Place of Presentation: 大阪
• [Presentation] がん細胞内環境を認識して遺伝子発現制御を行う高分子-ペプチドコンジュゲート (2008)
  • Author(s): 姜貞勲, ほか
  • Organizer: 第57回高分子討論会
  • Place of Presentation: 大阪
• [Presentation] 抗ウイルス薬開発の新戦略- ウイルス感染細胞特異的な遺伝子発現制御法の創出 (2008)
  • Author(s): 浅井大輔, ほか
  • Organizer: 第56回日本ウイルス学会
  • Place of Presentation: 岡山
• [Presentation] 細胞内シグナル応答型遺伝子キャリアーのソノポレーション法による高効率な細胞導入 (2008)
  • Author(s): 土谷享, ほか
  • Organizer: 日本バイオマテリアル学会シンポジウム2008
  • Place of Presentation: 東京
• [Presentation] 細胞内シグナル応答型遺伝子制御型キャリヤーの設計 (2008)
  • Author(s): 片山佳樹, ほか
  • Organizer: 日本バイオマテリアル学会シンポジウム2008
  • Place of Presentation: 東京
• [Book] 国際科学技術財團研究助成研究内容解説(疾患細胞選択的な遺伝子発現制御技術の創出と治療への応用(細胞と対話する技術の創出-薬物を病変細胞でだけ働かせる新しい考え方)) (2009)
  • Author(s): 浅井大輔
• [Remarks] 研究室ホームページ(研究業績等)
  • URL: http://www.marianna-u.ac.jp/microbiology/office/003347.html
• [Remarks] 国際科学技術財団研究助成
  • URL: http://www.japanprize.jp/subsidy_h20.html
• [Patent(Industrial Property Rights)] Rho-キナーゼの新規基質ペプチド (2009)
  • Inventor(s): 片山佳樹、姜貞勲、浅井大輔
  • Industrial Property Rights Holder: 片山佳樹、姜貞勲、浅井大輔
  • Industrial Property Number: 2009-211147
  • Filing Date: 2009
  • Acquisition Date: 2009
• [Patent(Industrial Property Rights)] 蛍光リガンド及び化学物質のアンドロゲン受容体への結合強度測定方法 (2008)
  • Inventor(s): 下東康幸、中井誠、浅井大輔、矢可部芳州
  • Industrial Property Rights Holder: 下東康幸、中井誠、浅井大輔、矢可部芳州
  • Filing Date: 2008
  • Acquisition Date: 2008