Protease-mediated regulation of host immune system and application as therapeutic targets

2009 Fiscal Year Final Research Report

• Project/Area Number: 19209058
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Kyushu University
• Principal Investigator: YAMAMOTO Kenji Kyushu University, 大学院・薬学研究院, 特任教授 (40091326)
• Co-Investigator(Kenkyū-buntansha): TSUKUBA Takayuki 長崎大学, 大学院・医歯薬学総合研究科, 教授 (30264055) ; TAKII Ryosuke 山口大学, 大学院・医学研究科, 助教 (00419558) ; KADOWAKI Tomoko 長崎大学, 大学院・医歯薬学総合研究科, 助教 (70336080) ; KAWAKUBO Tomoyo 九州大学, 大学院・薬学研究院, 特任助教 (70507813)
• Project Period (FY): 2007 – 2009
• Keywords: 歯学 / 蛋白質 / トランスレーショナルリサーチ / 発現制御 / 薬理学 / 癌 / 免疫学 / プロテアーゼ

Research Abstract

In this study, we investigated the nature and function of both host-and bacteria-derived proteases in host immune system. Cathepsin E (catE) and gingipains were treated as a host-derived endolysosomal protease and bacterial proteases, respectively. The obtained results are as follows : (i) CatE differentially regulated the nature and function of dendritic cells and macrophages, (ii) CatE deficiency caused impairment of autophagy in macrophages manifesting mitochondria dysfunction and enhanced oxidative stress, (iii) Endogenous Cat E expression levels were positively associated with the growth arrest and metastasis reduction of tumor cells in vivo and the concomitant increase of tumor-associated activated macrophages in tumor microenvironment, (iv) CatE specifically induced apoptosis in tumor cells without affecting normal cells in vitro by catalyzing the proteolytic release of soluble TRAIL from tumor cell surface, (v) Peptide-aptamer-based inhibitors and activators of CatE was generated by cDNA display techniques, (vi) Atherosclerosis progression in apolipoprotein E-knockout mice was accelerated by P. gingivalis infection, which was mediated by selective proteolysis of apolipoprotein B-100 by Arg-gingipain (Rgp), (vii) A risk for preterm birth and fetal death in pregnant mice was enhanced by P. gingivalis infection and inhibited by combination treatment of Rgp and Kgp inhibitors, (iii) A newly developed peptide analogue inhibiting both Rgp and Kgp suppressed most of the pethogenicity of P. gingivalis.

Research Products

• [Journal Article] Cathepsin E enhances anticancer activity of doxorubicin on human prostate cancer cells showing resistance to TRAIL-mediated apoptosis. (2010)
  • Author(s): Yasukochi A, Kawakubo T, Nakamura S, Yamamoto K.
  • Journal Title: Biol Chem (in press)
  • Peer Reviewed
• [Journal Article] Impaired chemotaxis and cell adhesion due to decrease in wide range of cell surface receptors in cathepsin E-deficient macrophages. (2009)
  • Author(s): Tsukuba T, Yanagawa M, Okamoto K, Okamoto Y, Yasuda Y, Nakayama KI, Kadowaki T, Yamamoto K.
  • Journal Title: J. Biochem. 145(5) Pages: 565-573
  • Peer Reviewed
• [Journal Article] Development of systemin in vitro evolution and its application to generation of peptide aptamer-based inhibitors of cathepsin E. (2009)
  • Author(s): Kitamura K, Yoshida C, Kinoshita Y, Kadowaki T, Takahashi Y, Tayama T, Kawakubo T, Naimuddin M, Salimulla M, Nemoto N, Hanada K, Husimi Y, Yamamoto K, Nishigaki K.
  • Journal Title: J. Mol. Biol. 387 Pages: 1186-1198
  • Peer Reviewed
• [Journal Article] Association of cathepsin E deficiency with the increased territorial aggressive response of mouse (2008)
  • Author(s): Shigematsu N., Fukuda T., Yamamoto T., Nishioku T., Yamaguchi T., Himeno M., Nakayama K.I., Tsukuba T., Kadowaki T., Okamoto K., Higuchi S., Yamamoto K.
  • Journal Title: J. Neurochem. 105 Pages: 1394-1404
  • Peer Reviewed
• [Journal Article] Determination of active site of Lysine-specific cysteine proteinase (Lys-gingipain) by use of a Porphyromonas gingivalis plasmid system. (2008)
  • Author(s): Ishida Y., Hu J.P., Sakai E., Kadowaki T., Yamamoto K., Tsukuba T., Nakayama K., Okamoto K.
  • Journal Title: Arch. Oral Biol. 53(6) Pages: 538-544
  • Peer Reviewed
• [Journal Article] An immunohistochemical study on a unique colocalization relationship between substance P and GABA in the central nucleus of amygdala. (2008)
  • Author(s): Shigematsu, N., Yamamoto, K., Higuchi, S., Fukuda, T.
  • Journal Title: Brain Res. 1198 Pages: 55-67
  • Peer Reviewed
• [Journal Article] Gene expression of profiling of mammary glands of cathepsin E-deficient mice compared with wild-type littermates. (2008)
  • Author(s): Kawakubo T., Yasukochi A., Tsukuba T., Kadowaki T., Yamamoto K.
  • Journal Title: Biochimie 90 Pages: 396-404
  • Peer Reviewed
• [Journal Article] Cathepsin E Prevents Tumor Growth and Metastasis by Catalyzing the Proteolytic Release of Soluble TRAIL from Tumor Cell (2007)
  • Author(s): Kawakubo T., Okamoto K., Iwata J., Shin M., Okamoto Y., Yasukochi A., Nakayama K.I., Kadowaki T., Tsukuba T., Yamamoto K.
  • Journal Title: Surface. Cancer Res. 67 Pages: 10869-10878
  • Peer Reviewed
• [Journal Article] Differential regulation of the nature and functions of dendritic cells and macrophages by cathepsin E. (2007)
  • Author(s): Kakehashi H., Nishioku T., Tsukuba T., Kadowaki T., Nakamura S., Yamamoto K.
  • Journal Title: J. Immunol. 179 Pages: 5728-5737
  • Peer Reviewed
• [Journal Article] Squamous cell carcinoma antigen 1 is an inhibitor of parasite-derived cysteine proteases. (2007)
  • Author(s): Kanaji, S., Tanaka, Y., Sakata, Y., Takeshita, K., Arima, K., Ohta, S., Hansell, E.J., Caffrey, C., Mottram, J.C., Lowther, J., Donnelly, S., Stack, C., Kadowaki, T., Yamamoto, K., McKerrow, J.H., Dalton, J.P., Coombs, G.H., Izuhara, K.
  • Journal Title: FEBS Lett. 581 Pages: 4260-4264
  • Peer Reviewed
• [Journal Article] Construction of recombinant hemagglutinin derived from the gingipain-encoding gene of Porphyromonas gingivalis, identification of its target protein on erthrocytes, and inhibition of hemaggelutination by an interdomain regional peptide. (2007)
  • Author(s): Sakai, E., Naito, M., Sato, K., Hotokezaka, H., Kadowaki, T., Kamaguch, A., Yamamoto, K., Okamoto, K., Nakayama, K.
  • Journal Title: J. Bacteriol. 189 Pages: 3977-3986
  • Peer Reviewed
• [Journal Article] Cathepsin E deficiency induces a novel form of lysosomal storage disorder 5 showing the accumulation of lysosomal membrane sialoglycoproteins and the elevation of lysosomal pH in macrophages. (2007)
  • Author(s): Yanagawa, M., Tsukuba, T., Nishioku, T., Okamoto, Y., Okamoto, K., Takii, R., Terada, Y., Nakayama, K.T., Kadowaki, T., Yamamoto, K
  • Journal Title: J. Biol. Chem. 282 Pages: 1851-1862
  • Peer Reviewed
• [Journal Article] Association of cathepsin E with tumor growth arrest through angiogenesis inhibition and enhanced immune responses. (2007)
  • Author(s): Shin, M., Kadowaki, T., Iwata, J., Kawakubo, T., Yamaguchi, N., Takii, R., Tsukuba, T., Yamamoto, K.
  • Journal Title: Biol. Chem. 388 Pages: 1173-1181
  • Peer Reviewed
• [Journal Article] A role for gingipains in cellular responses and bacterial survival in Porphyromonas gingicalis-infected cells. (2007)
  • Author(s): Kadowaki, T., Takii, R., Yamatake, K., Kawakubo, T., Tsukuba, T., Yamamoto, K.
  • Journal Title: Frontiers in Biosci. 12 Pages: 4800-4809
  • Peer Reviewed
• [Journal Article] Role for gingipains in Porphyromonas gingivalis traffic to phagolysosomes and survival in human aortic endothelilal cells. (2007)
  • Author(s): Yamatake, K., Maeda, M., Kadowaki, T., Takii, R., Tsukuba, T., Ueno, T., Kominami, E., Yokota, S., Yamamoto, K.
  • Journal Title: Infect. Immun. 75 Pages: 2090-2100
  • Peer Reviewed
• [Journal Article] Selection-by-function: effifient enrichment of cathepsin E inhibitors from a DNA library. (2007)
  • Author(s): Naimuddin, M., Kitamura, K., Kinoshita, Y., Honda-Takehashi, Y., Murakami, M., Ito, M., Yamamoto, K., Hanada, K., Husimi, Y., Nishigaki, K.
  • Journal Title: J. Mol. Recognit. 20 Pages: 58-68
  • Peer Reviewed
• [Patent(Industrial Property Rights)] 新規ペプチド及びその使用 (2010)
  • Inventor(s): 西垣功一、北村幸一郎、Madhu Biyani、二上雅恵、山本健二、川久保友世、安河内篤
  • Industrial Property Rights Holder: 西垣功一、北村幸一郎、Madhu Biyani、二上雅恵、山本健二、川久保友世、安河内篤
  • Industrial Property Number: 特許,特願2009-245763
  • Filing Date: 2010-03-17
  • Acquisition Date: 2010-03-17
• [Patent(Industrial Property Rights)] カテプシンEの腫瘍マーカーとしての用途及びカテプシンEならびにカテプシンDの腫瘍血管新生阻害療法のターゲットとしての用途 (2010)
  • Inventor(s): 山本健二、岩田淳一
  • Industrial Property Rights Holder: 山本健二、岩田淳一
  • Industrial Property Number: 特許,特許第4452791号
  • Filing Date: 2010-02-12
  • Acquisition Date: 2010-02-12
• [Patent(Industrial Property Rights)] アンチエイジング剤 (2009)
  • Inventor(s): 山本健二、川久保友世、安河内篤
  • Industrial Property Rights Holder: 山本健二、川久保友世、安河内篤
  • Industrial Property Number: 特許,特願2009-264894
  • Filing Date: 2009-11-20
  • Acquisition Date: 2009-11-20
• [Patent(Industrial Property Rights)] 新規な抗癌カテプシン製剤およびそれを用いた癌併用療法抗癌剤 (2007)
  • Inventor(s): 山本健二、川久保友世、安河内篤
  • Industrial Property Rights Holder: 山本健二、川久保友世、安河内篤
  • Industrial Property Number: 特許,特願2008-500586
  • Filing Date: 2007-02-19
  • Acquisition Date: 2007-02-19