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2022 Fiscal Year Research-status Report

Study the mechanism how Rab32/Rab38 positive lysosome related organelle is involved in bacteria suppression through macroautophagy and microautophagy in macrophage and mouse infectious model.

Research Project

Project/Area Number 20K15789
Research InstitutionOsaka University

Principal Investigator

LU SHIOULING  大阪大学, 大学院歯学研究科, 助教 (80830083)

Project Period (FY) 2020-04-01 – 2024-03-31
KeywordsGroup A Streptococcus / Macrophage / Rab32 / Rab38
Outline of Annual Research Achievements

In the pass year, I investigated whether Rab32/Rab38 is involved in macroautophagy against GAS.
Autophagic core proteins, ATGs, were important for macroautophagy induction even under GAS infection. It has been reported that LC3, an autophagosome marker, recruited to GAS in macrophage. I generated Atg7KO or FIP200KO macrophages generated by CRISPR Cas9 system, in which GFP-Rab32/Rab38 consist expressing. I found even without Atg7 or FIP200, Rab32 and Rab38 still strongly surrounding GAS under confocal microscope observation. LC3, an autophagy marker still recruit to GAS in macrophage, which indicated as an LC3 associated phagosome, instead of macroautophagy. I assume Rab32/38-mediated LAP, which containing lysosome fusion, is much important than macroautophagy in macrophage.
Furthermore, Rab32/38 double KO cells showed that GAS-phagosome are required with lysosome (LAMP1), however, acidification seems defective inside this phagosome.
Finally, I also performed EM images, as results showed that there are condensed intensity of lysosome-fused GAS-containing phagosome and a classical double membrane surrounding GAS in Wild-type macrophage. In contrast, a low intensity of GAS-containing phagosome, many free or empty space, was majorly observed in Rab32/Rab38 double KO macrophage. We conclude that Rab32/Rab38 is very important for LAP pathway, especially in acidification.

Current Status of Research Progress
Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

In the pass year, the experiments were progressing under try and error, and the results came out there is some difference between wild-type and Rab32/Rab38 double DO cells.

Strategy for Future Research Activity

In the new year, I aim to investigate whether microautophagy target GAS and whether Rab32/Rab38 is involved in this lysosome wrapping mechanism (LWM). This phenotype will required more clear EM images analysis. Lysosome has potential to change into a long-shapes in macrophage. This active movement, named lysosome protrusion, is able to surround an organelle bigger than lysosome itself. I plan to find out whether cytosol GAS can be surrounded by lysosome directly. The structure of lysosome protrusion along GAS will also be confirmed by correlative light and electron microscope (CLEM). I hope to show a direct evidence of microautophagy against bacteria.

  • Research Products

    (10 results)

All 2023 2022 Other

All Int'l Joint Research (2 results) Journal Article (3 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (5 results) (of which Int'l Joint Research: 3 results,  Invited: 1 results)

  • [Int'l Joint Research] 台湾 /Chang Gung University/Dept. of Microbiology and Immunology(その他の国・地域)

    • Country Name
      その他の国・地域
    • Counterpart Institution
      台湾 /Chang Gung University/Dept. of Microbiology and Immunology
  • [Int'l Joint Research] Indiana University Bloomington/Dept. of Chemistry(米国)

    • Country Name
      U.S.A.
    • Counterpart Institution
      Indiana University Bloomington/Dept. of Chemistry
  • [Journal Article] VEGF-Mediated Augmentation of Autophagic and Lysosomal Activity in Endothelial Cells Defends against Intracellular Streptococcus pyogenes2022

    • Author(s)
      Lu Shiou-Ling、Omori Hiroko、Zhou Yi、Lin Yee-Shin、Liu Ching-Chuan、Wu Jiunn-Jong、Noda Takeshi
    • Journal Title

      mBio

      Volume: 13 Pages: -

    • DOI

      10.1128/mbio.01233-22

    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] VEGF (vascular endothelial growth factor) provides antimicrobial effects via autophagy and lysosomal empowerment in endothelial cells2022

    • Author(s)
      Lu Shiou-Ling、Noda Takeshi
    • Journal Title

      Autophagy Reports

      Volume: 1 Pages: 555~558

    • DOI

      10.1080/27694127.2022.2137755

    • Peer Reviewed
  • [Journal Article] オートファジーは健康寿命に寄与するか?2022

    • Author(s)
      野田健司、Shiou-Ling Lu
    • Journal Title

      大阪歯科保険医新聞

      Volume: 1445 Pages: -

    • Open Access
  • [Presentation] How Streptococcus pyogenes is (or is not) targeted by autophagy in blood vessel endothelial cells.2023

    • Author(s)
      Shiou-Ling Lu
    • Organizer
      第96回日本細菌学会総会
  • [Presentation] Study of Rab32/Rab38-positive lysosome- related-organelle involved in GAS clearance in macrophage.2022

    • Author(s)
      Shiou-Ling Lu
    • Organizer
      EMBO | EMBL Symposium: Innate immunity in host-pathogen interactions.
    • Int'l Joint Research
  • [Presentation] VEGF-mediated augmentation of autophagic and lysosomal activity in endothelial cells defends against intracellular Streptococcus pyogenes.2022

    • Author(s)
      Shiou-Ling Lu
    • Organizer
      第20回あわじ感染と免疫国際フォーラム
    • Int'l Joint Research
  • [Presentation] Mitochondrial autophagy is carried out by microautophagy in macrophages.2022

    • Author(s)
      Shiou-Ling Lu
    • Organizer
      The 10th International Symposium on Autophagy.
    • Int'l Joint Research / Invited
  • [Presentation] Rab32とRab38は酸加水分解酵素とV-ATPaseの細胞内輸送を制御することにより破骨細胞の機能を促進する2022

    • Author(s)
      Lu Shiou-Ling
    • Organizer
      第95回日本生化学会大会

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Published: 2023-12-25  

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