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

2009 Fiscal Year Final Research Report

• 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
• 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

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] 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
  • 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] 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)
• [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
  • Year and Date: 20090520-20090523
• [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
  • Year and Date: 20090402-20090404
• [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] 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
  • Year and Date: 20081026-20081028
• [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
  • Year and Date: 20080826-20080829
• [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
  • Year and Date: 20080629-20080630
• [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
  • Total Pages: 9-30
• [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: 特許・特願2009-211147
  • Filing Date: 20090900
  • Acquisition Date: 20090900
• [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
  • Industrial Property Number: 特許・特許第4180882号
  • Filing Date: 20080900
  • Acquisition Date: 20080900