Molecular mechanism of drug tolerance in dormant mycobacteria and identification of the drug target

2011 Fiscal Year Final Research Report

• Project/Area Number: 21590486
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Osaka City University
• Principal Investigator: MATSUMOTO Sohkichi 大阪市立大学, 大学院・医学研究科, 准教授 (30244073)
• Co-Investigator(Kenkyū-buntansha): NIKI Mamiko 大阪市立大学, 大学院・医学研究科, 助教 (20438229)
• Project Period (FY): 2009 – 2011
• Keywords: 感染症 / 結核 / 薬剤耐性 / 増殖 / イソニアジド

Research Abstract

Tuberculosis is deadly infectious diseases found worldwide and remains a leading public health problem. Although isoniazid(INH) is a key drug for the treatment of tuberculosis(TB), tolerance against INH that causes prolonged treatment duration is a concern for effective TB chemotherapy. INH is a prodrug and is activated by the mycobacterial enzyme, KatG. We have shown that mycobacterial DNA-binding protein 1(MDP1), which is a histone-like protein conserved in mycobacteria, negatively regulates KatG transcription and leads to phenotypic tolerance to INH in mycobacteria. Mycobacterium smegmatis deficient for MDP1 exhibited increased expression of KatG and showed enhanced INH activation compared to the wild-type strain. Expression of MDP1 was increased in the stationary phase and conferred growth phase-dependent tolerance of M. smegmatis to INH. Regulation of KatG expression is conserved among M. smegmatis and Mycobacterium tuberculosis complex. Artificial reduction of MDP1 in Mycobacterium bovis BCG was shown to lead to increased KatG expression and susceptibility to INH. These data demonstrate the phenotypic tolerance mechanism to INH of mycobacteria by a histone-like protein.

Research Products

• [Journal Article] A novel mechanism of growth phase-dependent tolerance to isoniazid in mycobacteria (2012)
  • Author(s): Niki, M., Tateishi Y., Ozeki Y., Kirikae T, Lewin A., Inoue Y, Matsumoto M., Dahl JL., Ogura H., Kobayashi K., and Matsumoto S.
  • Journal Title: J Biol Chem in press
  • Peer Reviewed
• [Journal Article] Mitochondrial density contributes to the immune response of macrophages to lipopolysaccharide via the MAPK pathway (2011)
  • Author(s): Kasahara, E., A. Sekiyama, M. Hori, K. Hara, N. Takahashi, M. Konishi, E. F. Sato, S. Matsumoto, H. Okamura, and M. Inoue
  • Journal Title: FEBS Lett Volume: 585 Pages: 2263-2268
  • Peer Reviewed
• [Journal Article] Loss of anti-mycobacterial efficacy in mice over time following vaccination with Mycobacterium bovis bacillus Calmette-Guerin (2011)
  • Author(s): Ozeki, Y., Y. Hirayama, T. Takii, S. Yamamoto, K. Kobayashi, and S. Matsumoto
  • Journal Title: Vaccine Volume: 29 Pages: 6881-6887
  • Peer Reviewed
• [Journal Article] A Histone-Like Protein of Mycobacteria Possesses Ferritin Superfamily Protein-Like Activity and Protects against DNA Damage by Fenton Reaction (2011)
  • Author(s): Takatsuka, M., M. Osada-Oka, E. F. Satoh, K. Kitadokoro, Y. Nishiuchi, M. Niki, M. Inoue, K. Iwai, T. Arakawa, Y. Shimoji, H. Ogura, K. Kobayashi, A. Rambukkana, and S. Matsumoto
  • Journal Title: PLoS One Volume: 6 Pages: e20985
  • Peer Reviewed
• [Journal Article] Development of avian influenza virus H5 DNA vaccine and MDP-1 gene of Mycobacterium bovis as genetic adjuvant (2010)
  • Author(s): Jalilian, B., A. R. Omar, M. H. Bejo, N. B. Alitheen, M. Rasoli, and S. Matsumoto
  • Journal Title: Genet Vaccines Ther Volume: 8 Pages: 4
  • Peer Reviewed
• [Journal Article] Differential recruitment of CD63 and Rab7-interacting-lysosomal-protein to phagosomes containing Mycobacterium tuberculosis in macrophages (2010)
  • Author(s): Seto, S., S. Matsumoto, K. Tsujimura, and Y. Koide
  • Journal Title: Microbiol Immunol Volume: 54 Pages: 170-174
  • Peer Reviewed
• [Journal Article] Characterization of murine T-cell epitopes on mycobacterial DNA-binding protein 1(MDP1) using DNA vaccination (2010)
  • Author(s): Suzuki, D., T. Nagata, G. Eweda, S. Matsumoto, M. Matsumoto, K. Tsujimura, and Y. Koide
  • Journal Title: Vaccine Volume: 28 Pages: 2020-2025
  • Peer Reviewed
• [Journal Article] Controlled expression of branch-forming mannosyltransferase is critical for mycobacterial lipoarabinomannan biosynthesis (2010)
  • Author(s): Sena, C. B., T. Fukuda, K. Miyanagi, S. Matsumoto, K. Kobayashi, Y. Murakami, Y. Maeda, T. Kinoshita, and Y. S. Morita
  • Journal Title: J Biol Chem Volume: 285 Pages: 13326-13336
  • Peer Reviewed
• [Journal Article] Transient role of CD4+CD25+regulatory T cells in mycobacterial infection in mice (2010)
  • Author(s): Ozeki, Y., I. Sugawara, T. Udagawa, T. Aoki, M. Osada-Oka, Y. Tateishi, H. Hisaeda, Y. Nishiuchi, N. Harada, K. Kobayashi, and S. Matsumoto
  • Journal Title: Int Immunol Volume: 22 Pages: 179-189
  • Peer Reviewed
• [Journal Article] Growth inhibition of HeLa cell by internalization of Mycobacterium bovis Bacillus Calmette-Guerin(BCG) Tokyo (2009)
  • Author(s): Kitamura, A., S. Matsumoto, and I. Asahina
  • Journal Title: Cancer Cell Int Volume: 9 Pages: 30
  • Peer Reviewed
• [Journal Article] Mycobacterium avium complex organisms predominantly colonize in the bathtub inlets of patients' bathrooms (2009)
  • Author(s): Nishiuchi, Y., A. Tamura, S. Kitada, T. Taguri, S. Matsumoto, Y. Tateishi, M. Yoshimura, Y. Ozeki, N. Matsumura, H. Ogura, and R. Maekura
  • Journal Title: Jpn J Infect Dis Volume: 62 Pages: 182-186
  • Peer Reviewed
• [Journal Article] Mycobacterium kyorinense sp. nov., a novel, slow-growing species, related to Mycobacterium celatum, isolated from human clinical specimens (2009)
  • Author(s): Okazaki, M., K. Ohkusu, H. Hata, H. Ohnishi, K. Sugahara, C. Kawamura, N. Fujiwara, S. Matsumoto, Y. Nishiuchi, K. Toyoda, H. Saito, S. Yonetani, Y. Fukugawa, M. Yamamoto, H. Wada, A. Sejimo, A. Ebina, H. Goto, T. Ezaki, and T. Watanabe
  • Journal Title: Int J Syst Evol Microbiol Volume: 59 Pages: 1336-1341
  • Peer Reviewed
• [Journal Article] Dissection of Rab7 localization on Mycobacterium tuberculosis phagosome (2009)
  • Author(s): Seto, S., S. Matsumoto, I. Ohta, K. Tsujimura, and Y. Koide
  • Journal Title: Biochem Biophys Res Commun Volume: 387 Pages: 2722-277
  • Peer Reviewed
• [Journal Article] Virulence of Mycobacterium avium complex strains isolated from immune-competent patients (2009)
  • Author(s): Tateishi, Y., Y. Hirayama, Y. Ozeki, Y. Nishiuchi, M. Yoshimura, J. Kang, A. Shibata, K. Hirata, K. Kitada, S. Kitada, R. Maekura, H. Ogura, K. Kobayashi, and S. Matsumoto
  • Journal Title: Microb Pathog Volume: 46 Pages: 6-12
  • Peer Reviewed
• [Journal Article] High transmissibility of the modern Beijing Mycobacterium tuberculosis in homeless patients of Japan (2009)
  • Author(s): Wada, T., S. Fujihara, A. Shimouchi, M. Harada, H. Ogura, S. Matsumoto, and A. Hase
  • Journal Title: Tuberculosis(Edinb) Volume: 19 Pages: 19
  • Peer Reviewed
• [Journal Article] Mycobacteria Exploit Host Hyaluronan for Efficient Extracellular Replication (2009)
  • Author(s): T Hirayama, Y., M. Yoshimura, Y. Ozeki, I. Sugawara, T. Udagawa, S. Mizuno, N. Itano, K. Kimata, A. Tamaru, H. Ogura, K. Kobayashi, and S. Matsumoto
  • Journal Title: PLoS Pathog Volume: 5 Pages: e1000643
  • Peer Reviewed
• [Presentation] 結核菌の気道ヒアルロン酸を利用した感染と生体内増殖 (2010)
  • Author(s): 松本壮吉
  • Organizer: 第22回微生物シンポジウム
  • Place of Presentation: 大阪
  • Year and Date: 20100903-04
• [Presentation] Transient role of CD4^+CD25^+regulatory T cells in Mycobacterium tuberculosis infection in mice (2010)
  • Author(s): Y. Ozeki, I. Sugawara, T. Udagawa, H. Hisaeda, K. Kobayashi, and S. Matsumoto
  • Organizer: 14^<th> International Congress of Immunology
  • Place of Presentation: 神戸
  • Year and Date: 20100822-27
• [Presentation] Mycobacteria exploit host hyaluronan for efficient extracellular replication (2010)
  • Author(s): Sohkichi Matsumoto
  • Organizer: International Society for Hyaluronan Sciences 8th Internarional Congerence Hyaluronan
  • Place of Presentation: 京都
  • Year and Date: 20100606-11
• [Presentation] 抗酸菌感染症における感染制御の進歩 (2010)
  • Author(s): 松本壮吉
  • Organizer: 第84回日本感染症学会総会
  • Place of Presentation: 東京
  • Year and Date: 20100405-06
• [Presentation] 生活環境由来Mycobacterium avium complexの遺伝子多型解析 (2010)
  • Author(s): 西内由紀子、松本壮吉、立石善隆
  • Organizer: 第83回日本細菌学会総会
  • Place of Presentation: 横浜
  • Year and Date: 20100327-29
• [Presentation] 潜在性結核の診断法の確立に向けた臨床への橋渡し研究 (2010)
  • Author(s): 岡真優子、平山幸雄、立石善隆、小林和夫、松本壮吉
  • Organizer: 第1報、第83回日本細菌学会総会
  • Place of Presentation: 横浜
  • Year and Date: 20100327-29
• [Presentation] 結核菌感染における制御性T細胞の関与 (2010)
  • Author(s): 尾関百合子、菅原勇、岡真優子、小林和夫、松本壮吉
  • Organizer: 第83回日本細菌学会総会
  • Place of Presentation: 横浜
  • Year and Date: 20100327-29
• [Presentation] Molecular mechanisms of latent tuberculiosis infection(LTBI) and development of diagnosis of LTBI (2010)
  • Author(s): 松本壮吉
  • Organizer: 第3回日中科学フォーラム
  • Place of Presentation: 中国湖北省武漢市
  • Year and Date: 20100315-16
• [Presentation] 潜在性結核菌に対する液性免疫応答の解析 (2010)
  • Author(s): 岡真優子、平山幸雄、立石善隆、小林和夫、前倉亮治、松本壮吉
  • Organizer: 第8回感染症沖縄フォーラム
  • Place of Presentation: 沖縄
  • Year and Date: 20100211-13
• [Presentation] 感染マクロファージにおけるサイトカイン発現パターンの差異からみた肺MAC症の重症化機序の解明 (2010)
  • Author(s): 立石善隆、岡真優子、西内由紀子、小林和夫、前倉亮治、松本壮吉
  • Organizer: 第8回感染症沖縄フォーラム
  • Place of Presentation: 沖縄
  • Year and Date: 20100211-13
• [Presentation] 結核菌の長生きのメカニズム;休眠現象と潜在性結核 (2010)
  • Author(s): 松本壮吉
  • Organizer: 第27回東海薬物治療研究会
  • Place of Presentation: 名古屋
  • Year and Date: 2010-05-29
• [Presentation] A ferritin superfamily-like protein in mycobacteria (2009)
  • Author(s): Mayuko Osada-Oka, Masaki Takatsuka, F Eisuke Sato, Yuriko Ozeki, Mamiko Yoshimura, Masayasu Inoue, Kazuo Kobayashi, Sohkichi Matsumoto
  • Organizer: THE9TH AWAJI INTERNATIONAL FORUM
  • Place of Presentation: 淡路島
  • Year and Date: 20090908-11
• [Presentation] 肺Mycobacterium avium complex症患者由来M. aviumと豚由来M. aviumの血清型比較 (2009)
  • Author(s): 西内由紀子、田栗貴博、松本壮吉、立石善隆、北田清悟、田丸亜貴、鈴木定彦、前倉亮治
  • Organizer: 第84回日本結核病学会総会
  • Place of Presentation: 札幌
  • Year and Date: 20090702-03
• [Presentation] MLPA法による多剤耐性結核菌の検出 (2009)
  • Author(s): 田丸亜貴、河原隆二、松本壮吉、鈴木定彦
  • Organizer: 第84回日本結核病学会総会
  • Place of Presentation: 札幌
  • Year and Date: 20090702-03
• [Presentation] 集団感染事例におけるVNTR型とIS6110-RFLPパターンの比較 (2009)
  • Author(s): 田丸亜貴、松本壮吉
  • Organizer: 第84回日本結核病学会総会
  • Place of Presentation: 札幌
  • Year and Date: 20090702-03
• [Presentation] 抗結核菌薬スクリーニング系の確立と実践 (2009)
  • Author(s): 尾関百合子、原田誠、西内由紀子、山本三郎、小林和夫、松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] 抗酸菌の薬剤感受性におけるMDP1の調節機構の解析 (2009)
  • Author(s): 仁木誠、吉村満美子、小林和夫、松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] DNAワクチンを用いたMycobacterium DNA-binding protein 1(MDP1)のマウスT細胞エピトープの同定 (2009)
  • Author(s): 鈴木大介、永田年、辻村邦夫、松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] 結核菌感染マクロファージにおけるhypoxia-inducible factor-1alphaの作用機構 (2009)
  • Author(s): 岡真優子、小林和夫、松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] Mycobacterium intracellulare由来血清型7, 12, 13型糖ペプチド脂質の構造類似性とオリゴ糖解析 (2009)
  • Author(s): 藤原永年、中田登、中崇、水野淨子、合田麗奈、牧野正彦、吉村満美子、松本壮吉、前田伸司
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] 生活環境に分布するMycobacterium avium complexと臨床分離株の多型解析 (2009)
  • Author(s): 西内由紀子、松本壮吉、立石善隆、鈴木定彦
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] 結核菌の増殖に対するヒアルロン酸の作用 (2009)
  • Author(s): 平山幸雄、吉村満美子、尾関百合子、菅原勇、小林和夫、松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] 急速な臨床経過に合致した高病原性Mycobacterium avium-intracellulare complex菌株の同定 (2009)
  • Author(s): 立石善隆、平山幸雄、尾関百合子、西内由紀子、吉村満美子、小林和夫、松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] Tuberculosis, from biology to development of control strategies (2009)
  • Author(s): 松本壮吉
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋
  • Year and Date: 20090312-14
• [Presentation] 急速な臨床経過に合致した高病原性MAC菌株の同定-臨床病態と細菌学的病原性との関連 (2009)
  • Author(s): 立石善隆、岡真優子、西内由紀子、吉村満美子、小林和夫、前倉亮治、松本壮吉
  • Organizer: 第7回感染症沖縄フォーラム
  • Place of Presentation: 沖縄
  • Year and Date: 20090212-14
• [Presentation] 抗酸菌の休眠誘導と遺伝子発現解析 (2009)
  • Author(s): 吉村満美子、仁木誠、岡真優子、尾関百合子、原田誠、田丸亜貴、立石善隆、西内由紀子、小林和夫、松本壮吉
  • Organizer: 第7回感染症沖縄フォーラム
  • Place of Presentation: 沖縄
  • Year and Date: 20090212-14
• [Presentation] Mycobacterial DNA-binding protein-1の新機能-鉄の酸化活性と貯蔵性- (2009)
  • Author(s): 岡真優子、高塚正樹、尾関百合子、吉村満美子、小林和夫、松本壮吉
  • Organizer: 第7回感染症沖縄フォーラム
  • Place of Presentation: 沖縄
  • Year and Date: 20090212-14
• [Presentation] 低酸素状態でのHIF-1a安定化におけるグルコースの作用機構 (2009)
  • Author(s): 岡真優子、今岡進、小林和夫、松本壮吉
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-10
• [Presentation] 休眠時における抗酸菌の遺伝子発現解析 (2009)
  • Author(s): 吉村満美子、仁木誠、立石善隆、小林和夫、松本壮吉
  • Organizer: 第62回日本細菌学会関西支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2009-11-14
• [Presentation] 抗酸菌は宿主ヒアルロン酸を利用して細胞外増殖を行う (2009)
  • Author(s): 平山幸雄、吉村満美子、尾関百合子、田丸亜貴、小倉壽、小林和夫、松本壮吉
  • Organizer: 第62回日本細菌学会関西支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2009-11-14
• [Presentation] 結核菌感染における制御性T細胞の役割検討 (2009)
  • Author(s): 尾関百合子、菅原勇、岡真優子、小林和夫、松本壮吉
  • Organizer: 第62回日本細菌学会関西支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2009-11-14
• [Presentation] 気道ヒアルロン酸を利用した、結核菌の感染と増殖のストラテジー (2009)
  • Author(s): 松本壮吉
  • Organizer: 第50回日本熱帯医学学会大会
  • Place of Presentation: 沖縄
  • Year and Date: 2009-10-22
• [Presentation] MDP1タンパクのマウスT細胞エピトープの解析/Characterization of murine T-cell epitopes in Mycobacterial DNAbinding protein 1 (2008)
  • Author(s): SUZUKI Daisuke, NAGATA Toshi, MATSUMOTO Sohkichi, KOIDE Yukiko
  • Organizer: 第38回日本免疫学会総会
  • Place of Presentation: 京都
  • Year and Date: 20081201-03
• [Presentation] Proteciton of DNA by mycobacterial DNA-binding protein 1(MDP1) by preventing the ironinduced Fenton reaction (2008)
  • Author(s): MATSUMOTO Sohkichi
  • Organizer: 43th US-Japan, Cooperative Medical Science Program
  • Place of Presentation: USA
  • Year and Date: 20080729-31
• [Presentation] Mycobacterial DNA-binding protein 1(MDP1)に見いだされたFerritin-super family蛋白質様活性 (2008)
  • Author(s): 松本壮吉、尾関百合子、吉村満美子、藤原永年、西内由紀子、立石善隆、小林和夫
  • Organizer: 第61回日本細菌学会関西支部総会
  • Place of Presentation: 京都
  • Year and Date: 2008-11-08
• [Presentation] 抗酸菌の薬剤感受性におけるMDP1の調節機構の解析 (2008)
  • Author(s): 仁木誠、吉村満美子、松本壮吉、和田崇之、小林和夫
  • Organizer: 第83回日本結核病学会
  • Place of Presentation: 東京
  • Year and Date: 2008-04-24
• [Presentation] ヒアルロン酸の結核病巣における産生と局在 (2008)
  • Author(s): 平山幸雄、吉村満美子、尾関百合子、菅原勇、青木俊明、西内由紀子、小林和夫
  • Organizer: 第83回日本結核病学会
  • Place of Presentation: 東京
  • Year and Date: 2008-04-24
• [Presentation] Mycobacterial DNA-binding protein 1(MDP1)による増殖と細胞壁合成の同調メカニズム (2008)
  • Author(s): 松本壮吉、尾関百合子、西内由紀子、藤原永年、吉村満美子、小林和夫
  • Organizer: 第83回日本結核病学会
  • Place of Presentation: 東京
  • Year and Date: 2008-04-24
• [Book] 人獣共通感染症、木村哲、喜田宏編集、非結核性抗酸菌症改訂版 (2011)
  • Author(s): 西内由紀子、立石善隆、山田毅、松本壮吉
  • Publisher: 医薬ジャーナル社
• [Book] Host and bacterial factors that regulate Mycobacterium tuberculosis infection and persistence.(Yamamoto S, Maeyama J, and Takii T editors) (2011)
  • Author(s): Niki M and Matsumoto S
  • Publisher: Japan anti-tuberculosis association
• [Book] 分子生物学から見た結核研究の現在、四元秀毅、倉島篤行、結核Up to Date改訂3版四元秀毅、倉島篤行編集 (2010)
  • Author(s): 松本壮吉
  • Total Pages: 183-189
  • Publisher: 南江堂
• [Patent(Industrial Property Rights)] 免疫刺激G9.1の抗結核ブースターワクチン創出への応用 (2011)
  • Inventor(s): 伊保澄子、前山順一、松本壮吉、山本三郎
  • Industrial Property Rights Holder: 国立大学法人福井大学、国立感染症研究所、公立大学法人大阪市立大学、日本ビーシージー製造株式会社
  • Industrial Property Number: 用途特許、特願2011-080646
  • Filing Date: 2011-03-31
• [Patent(Industrial Property Rights)] MDP1による微生物を凝集および/または沈殿させる方法 (2009)
  • Inventor(s): 松本壮吉、山本法明
  • Industrial Property Rights Holder: コニカミノルタホールディングス株式会社
  • Industrial Property Number: 用途特許、PCT/JP2009/061819
  • Filing Date: 2009-06-29
  • Overseas
• [Patent(Industrial Property Rights)] 遅発育性抗酸菌ポリペプチド (2009)
  • Inventor(s): 山田毅、松本壮吉
  • Industrial Property Rights Holder: 大塚製薬株式会社、山田毅、松本壮吉
  • Industrial Property Number: 物質特許、特許第4415200号
  • Acquisition Date: 2009-12-04