Role of extracellular vesicles as communication mediators between pathogens to potentiate pathogenicity in wound infections

• 研究課題/領域番号: 22K15454
• 研究種目: 若手研究
• 配分区分: 基金
• 審査区分: 小区分49050:細菌学関連
• 研究機関: 弘前大学
• 研究代表者: Subsomwong Phawinee 弘前大学, 医学研究科, 助教 (00883745)
• 研究期間 (年度): 2022-04-01 – 2025-03-31
• 研究課題ステータス: 交付 (2023年度)
• 配分額: 4,030千円 (直接経費: 3,100千円、間接経費: 930千円); 2024年度: 1,170千円 (直接経費: 900千円、間接経費: 270千円); 2023年度: 1,170千円 (直接経費: 900千円、間接経費: 270千円); 2022年度: 1,690千円 (直接経費: 1,300千円、間接経費: 390千円)
• キーワード: Staphylococcus aureus / Pseudomonas aeruginosa / Extracellular vesicles / Extracellular vesicle / Interaction / Pathogenicity

研究開始時の研究の概要

Chronic wound infections, especially by multidrug-resistant organism, becomes a challenge for treatment. Staphylococcus aureus and Pseudomonas aeruginosa are commonly found in chronic wounds. The synergistic effects of these two bacteria were demonstrated. However, their mediators are not yet fully reported. Here, we elucidate the role of extracellular vesicles (EVs) as mediators in the pathogenicity of S. aureus and P. aeruginosa. Understanding these mediators’ functions will be useful for applying the EVs as a target tool for the treatment of chronic wound infections.

研究実績の概要

In FY2022-23, we investigated the effect of EVs derived from S. aureus (SaEVs) and P. aeruginosa (PaEVs) on the pathogenicity of their bacterial counterparts. Our findings revealed that SaEVs had no impact on the growth and antibiotic pattern of P. aeruginosa. Surprisingly, SaEVs were found to enhance LPS production, biofilm formation, and the expression of polysaccharide polymerization-related genes in P. aeruginosa after SaEV treatment. Additionally, SaEVs increased the epithelial cell invasion of P. aeruginosa. Conversely, the uptake of P. aeruginosa by RAW264.7 macrophages was impaired after pretreatment with SaEVs. Regarding the inhibitory effect of PaEVs on S. aureus growth, we studied the mechanism in more detail. Our result revealed a reduction in lactate dehydrogenase 2 and formate acetyltransferase enzyme expression in the pyruvate fermentation pathway of S. aureus after PaEV treatment at both gene and protein levels. Furthermore, this inhibitory effect of PaEVs was abolished by supplementation with pyruvate or oxygen. It indicates that PaEVs inhibit the growth of S. aureus by suppressing the pyruvate fermentation pathway. In a further study, we plan to analyze the molecules in SaEVs and PaEVs that affect the P. aeruginosa pathogenicity and S. aureus growth. The EVs will be sequentially treated with enzymes including lipase, DNase, RNase, and protease. The activity of enzyme-treated EVs will be used to detect the promotion of P. aeruginosa pathogenicity and the inhibition of S. aureus growth.

現在までの達成度 (区分)

1: 当初の計画以上に進展している

理由

In FY2022-23, PI planed to isolate and purify the extracellular vesicles derived from S. aureus (SaEVs), and P. aeruginosa (PaEVs) culture supernatant. The effects of SaEVs and PaEVs on their respective bacterial counterparts were investigated. PI successfully isolated EVs and studied their effect on bacterial counterparts. SaEVs were found to enhance P. aeruginosa pathogenicity by increasing lipopolysaccharide biosynthesis, promoting biofilm formation, epithelial cell invasion, and impairing macrophage uptake. The mechanism of PaEVs in inhibiting S. aureus growth involved reducing lactate dehydrogenase 2 and formate acetyltransferase enzymes in the pyruvate fermentation pathway. This effect was abolished by supplementing with pyruvate or oxygen, indicating that PaEVs inhibit S. aureus growth by suppressing the pyruvate fermentation pathway. Finally, the PI and colleagues successfully published these two studies in international journals. Therefore, the PI self-evaluates as (1) Progressing More Smoothly Than Initially Planned.

今後の研究の推進方策

In FY2023, we observed that SaEVs enhance P. aeruginosa pathogenicity by increasing lipopolysaccharide biosynthesis, promoting biofilm formation, epithelial cell invasion, and impairing macrophage uptake. The mechanism of PaEVs in inhibiting S. aureus involved reducing lactate dehydrogenase 2 and formate acetyltransferase enzymes in the pyruvate fermentation pathway. To identify the active molecules of SaEVs that promote P. aeruginosa pathogenesis and PaEVs that inhibit S. aureus growth. The PI plans to sequentially treat the SaEVs and PaEVs with lipase, DNase, RNase, and protease. After treatment with each enzyme, the activity of SaEVs and PaEVs will be used to detect the ability to promote pathogenicity and inhibit growth in P. aeruginosa and S. aureus, respectively.

研究成果

• [雑誌論文] Heat Shock Protein SSA1 Enriched in Hypoxic Secretome of Candida albicans Exerts an Immunomodulatory Effect via Regulating Macrophage Function (2024)
  • 著者名/発表者名: Teng Wei、Subsomwong Phawinee、Narita Kouji、Nakane Akio、Asano Krisana
  • 雑誌名: Cells 巻: 13 号: 2 ページ: 127-127
  • DOI: 10.3390/cells13020127
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] Extracellular vesicles from Staphylococcus aureus promote the pathogenicity of Pseudomonas aeruginosa (2024)
  • 著者名/発表者名: Subsomwong Phawinee、Teng Wei、Ishiai Takahito、Narita Kouji、Sukchawalit Rojana、Nakane Akio、Asano Krisana
  • 雑誌名: Microbiological Research 巻: 281 ページ: 127612-127612
  • DOI: 10.1016/j.micres.2024.127612
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] The new 222‐nm far ultraviolet‐C lowers bacterial contamination to endoscopists during esophagogastroduodenoscopy (2023)
  • 著者名/発表者名: Fukutoku, Y., Kikuchi, H., Hoshi, K., Narita, K., Asari, T., Miyazawa, K., Sawada, Y., Hayamizu, S., Tatsuta, T., Oota, S., Hasui, K., Hiraga, H., Chinda, D., Mikami, T., Subsomwong, P., Asano, K., Yamane, K., Ogawa, Y., Sasaki, M., Koi, T., … Sakuraba, H.
  • 雑誌名: DEN Open 巻: 4 号: 1
  • DOI: 10.1002/deo2.292
  • 査読あり / オープンアクセス
• [雑誌論文] Genetic determinants of Biofilm formation of Helicobacter pylori using whole-genome sequencing (2023)
  • 著者名/発表者名: Fauzia Kartika Afrida、Aftab Hafeza、Miftahussurur Muhammad、Waskito Langgeng Agung、Tuan Vo Phuoc、Alfaray Ricky Indra、Matsumoto Takashi、Yurugi Michiyuki、Subsomwong Phawinee、Kabamba Evariste Tshibangu、Akada Junko、Yamaoka Yoshio
  • 雑誌名: BMC Microbiology 巻: 23 号: 1 ページ: 159-159
  • DOI: 10.1186/s12866-023-02889-8
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] Extracellular vesicles of Pseudomonas aeruginosa downregulate pyruvate fermentation enzymes and inhibit the initial growth of Staphylococcus aureus (2023)
  • 著者名/発表者名: Ishiai Takahito、Subsomwong Phawinee、Narita Kouj、Kawai Noriaki、Teng Wei、Suzuki Sachio、Sukchawalit Rojana、Nakane Akio、Asano Krisana
  • 雑誌名: Current Research in Microbial Sciences 巻: 4 ページ: 100190-100190
  • DOI: 10.1016/j.crmicr.2023.100190
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] Differential proteomic analysis and pathogenic effects of outer membrane vesicles derived from Acinetobacter baumannii under normoxia and hypoxia (2023)
  • 著者名/発表者名: Suzuki S, Subsomwong P, Narita K, Kawai N, Ishiai T, Teng W, Sukchawalit R, Nakane A, Tasaka S, Asano K.
  • 雑誌名: PLoS One 巻: 18(3) 号: 3 ページ: e0283109-e0283109
  • DOI: 10.1371/journal.pone.0283109
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] Mutations Related to Antibiotics Resistance in Helicobacter pylori Clinical Isolates from Bangladesh. (2023)
  • 著者名/発表者名: Fauzia, Kartika Afrida, Hafeza Aftab, Evariste Tshibangu-Kabamba, Ricky Indra Alfaray, Batsaikhan Saruuljavkhlan, Alain Cimuanga-Mukanya, Takashi Matsumoto, Phawinee Subsomwong, Junko Akada, Muhammad Miftahussurur, and Yoshio Yamaoka.
  • 雑誌名: Antibiotics 巻: 12 号: 2 ページ: 279-279
  • DOI: 10.3390/antibiotics12020279
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] Bacterial exposure risk to the endoscopist's face while performing endoscopy (2023)
  • 著者名/発表者名: Hoshi K, Kikuchi H, Narita K, Fukutoku Y, Asari T, Miyazawa K, Murai Y, Sawada Y, Tatsuta T, Hasui K, Hiraga H, Chinda D, Mikami T, Subsomwong P, Asano K, Nakane A, Fukuda S, Sakuraba H
  • 雑誌名: DEN Open 巻: 3(1) 号: 1
  • DOI: 10.1002/deo2.209
  • 査読あり / オープンアクセス
• [雑誌論文] Next-Generation Sequencing-Based Study of Helicobacter pylori Isolates from Myanmar and Their Susceptibility to Antibiotics (2022)
  • 著者名/発表者名: Subsomwong Phawinee、Doohan Dalla、Fauzia Kartika Afrida、Akada Junko、Matsumoto Takashi、Yee Than Than、Htet Kyaw、Waskito Langgeng Agung、Tuan Vo Phuoc、Uchida Tomohisa、Matsuhisa Takeshi、Yamaoka Yoshio
  • 雑誌名: Microorganisms 巻: 10 号: 1 ページ: 196-196
  • DOI: 10.3390/microorganisms10010196
  • 査読あり / オープンアクセス / 国際共著
• [学会発表] Staphylococcus aureus-derived extracellular vesicles promote pathogenicity of Pseudomonas aeruginosa (2023)
  • 著者名/発表者名: Subsomwong Phawinee
  • 学会等名: 第67回日本ブドウ球菌研究会
• [学会発表] Extracellular vesicles of Staphylococcus aureus promote pathogenicity of Pseudomonas aeruginosa (2023)
  • 著者名/発表者名: Subsomwong Phawinee
  • 学会等名: 第96回日本細菌学会総会
• [学会発表] Enhancement effector vesicles derived from Staphylococcus aureus on virulence of Pseudomonas aeruginosa (2022)
  • 著者名/発表者名: Subsomwong Phawinee
  • 学会等名: International Union of Microbiological Societies 2022 Congress
  • 国際学会
• [学会発表] Role of Staphylococcus aureus-derived extracellular vesicles in pathogenicity of Pseudomonas aeruginosa (2022)
  • 著者名/発表者名: Subsomwong Phawinee
  • 学会等名: 第74回日本細菌学会東北支部会
• [学会発表] Extracellular vesicles of Staphylococcus aureus promote exopolysaccharide biosynthesis and pathogenicity of Pseudomonas aeruginosa (2022)
  • 著者名/発表者名: Subsomwong Phawinee
  • 学会等名: 16回Wakate Colosseum
• [図書] Staphylococcus aureus Interplay between Bacteria and Hosts (Chapter 5; Immune Response to Staphylococcus aureus (2024)
  • 著者名/発表者名: Akio Nakane and Phawinee Subsomwong (Chapter 5)
  • 総ページ数: 210
  • 出版者: Springer
  • ISBN: 9789819994274