Development of rapid diagnosis of drug sensitivity and optimal choice of chemotherapeutic regimen in MAC pulmonary disease by making use of TnSeq

• Project/Area Number: 21K19637
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 58:Society medicine, nursing, and related fields
• Research Institution: Niigata University
• Principal Investigator: Yoshitaka Tateishi 新潟大学, 医歯学系, 准教授 (30433296)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2022: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2021: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
• Keywords: 非結核性抗酸菌症 / トランスポゾン / 次世代シーケンシング / 非結核性抗酸菌

Outline of Research at the Start

肺MAC症は中高年女性に急増する難治性の呼吸器感染症であるが、菌の遅発育性によりリアルタイムの病態把握と時期を得た治療薬の選択が困難であった。今回、トランスポゾン変異株作製と次世代シーケンシング技術を組み合わせたトランスポゾンシーケンシング（TnSeq）により、迅速かつゲノムワイドな薬剤感受性判定法の確立を目指す。TnSeqによる薬剤感受性判定法を確立できれば、起因菌株の生存必須遺伝子に基づいた治療薬剤の選択が可能となり、肺MAC症に対する「テーラーメード医療」への基盤構築につながる。

Outline of Final Research Achievements

For the purpose of developing the rapid diagnosis of drug sensitivity and optimal choice of chemotherapeutic regimen in MAC pulmonary disease, we have performed transposon sequencing of 7 clinical MAC strains and identified the common essential genes among the clinical strains. We identified genes of malate synthase (glcB) and type VII secretion system (eccC, eccB, eccB) as common essential genes, as well as genes of constitutive drug targets such as gyrA-gyrB, topA, embB, atpA-atpG-atpD, inhA, alr and rpoC.
We have analyzed the effect of monotherapy and combination therapy for mice infected with the high virulence strain. Clarithromycin-containing chemotherapy showed the highest efficacy. We could establish the mouse model of MAC pulmonary disease for evaluating in vivo drug sensitivity.

Academic Significance and Societal Importance of the Research Achievements

肺MAC症患者由来の臨床菌株を使ったトランスポゾンシーケンシング（TnSeq）により、現行治療薬の標的以外にも肺MAC病原体の治療標的が存在することが分かりました。今回の結果は、肺MAC症の新規治療法を開拓するための薬剤標的データベースとなります。
また、感染マウスモデルを使った生体内での治療効果判定手法を確立しました。これにより、将来開発される薬剤に対する生体内治療効果判定が可能となります。

Research Products

• [Journal Article] Virulence of Mycobacterium intracellulare clinical strains in a mouse model of lung infection - role of neutrophilic inflammation in disease severity. (2023)
  • Author(s): Tateishi Y, Ozeki Y, Nishiyama A, Miki M, Maekura R, Kida H, Matsumoto S
  • Journal Title: BMC Microbiol Volume: 23 Issue: 1 Pages: 94-94
  • DOI: 10.1186/s12866-023-02831-y
  • Peer Reviewed / Open Access
• [Journal Article] Lysocin E Targeting Menaquinone in the Membrane of Mycobacterium tuberculosis Is a Promising Lead Compound for Antituberculosis Drugs. (2022)
  • Author(s): Geberetsadik G, Inaizumi A, Nishiyama A, Yamaguchi T, Hamamoto H, Panthee S, Tamaru A, Hayatsu M, Mizutani Y, Kaboso SA, Hakamata M, Ilinov A, Ozeki Y, Tateishi Y, Sekimizu K, Matsumoto S.
  • Journal Title: Antimicrob Agents Chemother Volume: 66 Issue: 9
  • DOI: 10.1128/aac.00171-22
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Long-Term Prognosis and Antimycobacterial Glycolipid Antibody as Biomarker in Mycobacterium avium-intracellulare Complex Pulmonary Disease. (2022)
  • Author(s): Maekura R, Miki K, Tateishi Y, Matsumoto S, Kitada S, Miki M, Kida H.
  • Journal Title: Microbiol Spectr Volume: 10 Issue: 3
  • DOI: 10.1128/spectrum.00530-22
  • Peer Reviewed / Open Access
• [Journal Article] Monitoring IgG against Mycobacterium tuberculosis proteins in an Asian elephant cured of tuberculosis that developed from long-term latency. (2022)
  • Author(s): Ishikawa S, Ozeki Y, Suga S, Mukai Y, Kobayashi H, Inouchi E, Kaboso SA, Gebretsadik G, Dewi DNSS, Nishiyama A, Tateishi Y, Takihara H, Okuda S, Yoshida S, Misawa N, Matsumoto S.
  • Journal Title: Sci Rep Volume: 12 Issue: 1 Pages: 4310-4310
  • DOI: 10.1038/s41598-022-08228-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification and functional analysis of a new type of Z,E-mixed prenyl reductase from mycobacteria (2022)
  • Author(s): Abe, T., Hakamata, M., Nishiyama, A., Tateishi, Y., Matsumoto, S., Hemmi, H., Ueda, D., and Sato, T.
  • Journal Title: The FEBS journal Volume: - Issue: 16 Pages: 4981-4997
  • DOI: 10.1111/febs.16412
  • Peer Reviewed / Open Access
• [Journal Article] Direct attachment with erythrocytes augments extracellular growth of pathogenic Mycobacteria (2022)
  • Author(s): Nishiuchi, Y., Tateishi, Y., Hirano, H., Ozeki, Y., Yamaguchi, T., Miki, M., Kitada, S., Maruyama, F., Matsumoto, S.
  • Journal Title: Microbiology Spectrum Volume: in press Issue: 2
  • DOI: 10.1128/spectrum.02454-21
  • Peer Reviewed / Open Access
• [Journal Article] Extracellular DNA of slow growers of mycobacteria and its contribution to biofilm formation and drug tolerance. (2021)
  • Author(s): Ilinov A, Nishiyama A, Namba H, Fukushima Y, Takihara H, Nakajima C, Savitskaya A, Gebretsadik G, Hakamata M, Ozeki Y, Tateishi Y, Okuda S, Suzuki Y, Vinnik YS, Matsumoto S.
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 10953-10953
  • DOI: 10.1038/s41598-021-90156-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparative genomic analysis of Mycobacterium intracellulare: implications for clinical taxonomic classification in pulmonary Mycobacterium avium-intracellulare complex disease. (2021)
  • Author(s): Tateishi Y, Ozeki Y, Nishiyama A, Miki M, Maekura R, Fukushima Y, Nakajima C, Suzuki Y, Matsumoto S.
  • Journal Title: BMC Microbiol. Volume: 21 Issue: 1 Pages: 103-103
  • DOI: 10.1186/s12866-021-02163-9
  • Peer Reviewed / Open Access
• [Presentation] Mycobacterium intracellulare菌株間における生存必須遺伝子プロファイルの比較 (2023)
  • Author(s): 立石 善隆, 尾関 百合子, 西山 晃史, 松本 壮吉
  • Organizer: 第96回日本細菌学会総会
• [Presentation] 非結核性抗酸菌Mycobacterium intracellulare臨床菌株の実験的病原性および薬剤治療効果の検討 (2022)
  • Author(s): 立石 善隆, 尾関 百合子, 西山 晃史, 松本 壮吉
  • Organizer: 第59回日本細菌学会中部支部総会
• [Presentation] 非結核性抗酸菌臨床菌株におけるゲノムワイドな遺伝子必須性の相違と低酸素バイオフィルム形成との関係 (2022)
  • Author(s): 立石 善隆, 松本 壮吉
  • Organizer: 第35回日本バイフォルム学
• [Presentation] Mycobacterium intracellulare菌株間における生存必須遺伝子プロファイルの比較 (2022)
  • Author(s): 立石善隆、西山晃史、尾関百合子、松本壮吉
  • Organizer: 第95回日本細菌学会総会
• [Presentation] 結核・非結核性抗酸菌における バイオフィルム (2021)
  • Author(s): 立石善隆
  • Organizer: 第35回日本バイオフィルム学会
  • Invited
• [Presentation] Mycobacterium intracellulare臨床菌株の比較ゲノム解析による類縁菌種との異同およびゲノム多様性についての検討 (2021)
  • Author(s): 立石善隆、西山晃史、尾関百合子、松本壮吉.
  • Organizer: 第34バイオフィルム学会、第57日本細菌学会中部支部会.