Ryohei Yanoshita
Graduate School of Information Sciences,Major of Information Sciences | Dean of Graduate School/Chair of Program/Professor |
Faculty of Pharmaceutical Sciences,Department of Pharmaceutical Sciences | Professor |
Last Updated :2025/10/07
■Researcher basic information
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Educational Background
■Research activity information
Paper
- Distinguishing two distinct types of salivary extracellular vesicles: a potential tool for understanding their pathophysiological roles.
Yuko Ogawa; Yuri Miura; Mamoru Ikemoto; Atsushi Ohnishi; Yoshikuni Goto; Kazuma Aoki; Yuki Motokurumada; Yoshihiro Akimoto; Tamao Endo; Masafumi Tsujimoto; Ryohei Yanoshita
Frontiers in molecular biosciences, 2024
Extracellular vesicles (EVs), which are found in almost all cells and human body fluids, are currently being studied as a source of pathophysiological information. Previously, we demonstrated that at least two types of EVs can be isolated from human whole saliva (WS) using enzymatic activity of dipeptidyl peptidase IV (DPP IV) as a marker for differentiating the EV subsets. In the present study, EV fractions, termed EV-I 20 k-ppt and EV-II 100 k-ppt, were prepared by a combination of size-exclusion chromatography of improved condition and sequential centrifugation. The EV-I 20 k-ppt fraction contained medium/large EVs with a diameter of 100-1,000 nm, including aminopeptidase N (APN), mucin 1, ezrin, and Annexin A1. EV-II 100 k-ppt contained small EVs with a diameter of 20-70 nm, with DPP IV and CD9, programmed cell death 6-interacting protein, and tumor susceptibility gene 101 as characteristic proteins. Proteomic analyses also revealed distinctive repertoires of constituent proteins. Immunoprecipitation of several membrane proteins of the EVs with respective antibodies suggested their differential local membrane environment between the two types of salivary vesicles. Thus, we identified two distinctive types of EVs, one is APN/MUC1- rich EVs (EV-I, large/medium EVs) and the other is DPP IV/CD9-rich EVs (EV-II, small EVs). Furthermore, analysis of the binding of the EVs to coronavirus spike proteins showed that EV-II 100 k-ppt, but not EV-I 20 k-ppt, significantly bound to the spike protein of Middle East respiratory syndrome coronavirus (MERS-CoV). Finally, we developed a simple method to prepare two distinctive EVs from only 1 mL of human WS using sequential immunoprecipitation. Elucidating the features and functions of these two types of salivary EVs may help us understand their pathophysiological roles in the oral cavity and gastrointestinal tract. - Stability of human salivary extracellular vesicles containing dipeptidyl peptidase IV under simulated gastrointestinal tract conditions
Yuko Ogawa; Yoshihiro Akimoto; Mamoru Ikemoto; Yoshikuni Goto; Anna Ishikawa; Sakura Ohta; Yumi Takase; Hayato Kawakami; Masafumi Tsujimoto; Ryohei Yanoshita
Biochemistry and Biophysics Reports, Sep. 2021
BACKGROUND: Extracellular vesicles (EVs) have been isolated from various sources, including primary and cultured cell lines and body fluids. Previous studies, including those conducted in our laboratory, have reported the stability of EVs under various storage conditions. METHODS: EVs from human whole saliva were separated via size-exclusion chromatography. To simulate the effects of gastric or intestinal fluids on the stability of EVs, pepsin or pancreatin was added to the samples. Additionally, to determine the effect of bile acids, sodium cholate was added. The samples were then subjected to western blotting, dynamic light scattering, and transmission electron microscopy analyses. In addition, the activity of dipeptidyl peptidase (DPP) IV retained in the samples was examined to monitor the stability of EVs. RESULTS: Under acidic conditions, with pepsin mimicking the milieu of the stomach, the EVs remained stable. However, they partially lost their membrane integrity in the presence of pancreatin and sodium cholate, indicating that they may be destabilized after passing through the duodenum. Although several associated proteins, such as mucin 5B and CD9 were degraded, DPP IV was stable, and its activity was retained under the simulated gastrointestinal conditions. CONCLUSION: Our data indicate that although EVs can pass through the stomach without undergoing significant damage, they may be disrupted in the intestine to release their contents. The consistent delivery of active components such as DPP IV from EVs into the intestine might play a role in the efficient modulation of homeostasis of the signal transduction pathways occurring in the gastrointestinal tract. - ヒト唾液由来細胞外小胞の胃から腸内における条件での安定性の検討
Mar. 2021 - ヒト唾液由来exosomeの表面分子の結合状態および免疫活性化の制御への関与
Nov. 2018 - Exosome-associated Shiga toxin 2 is released from cells and causes severe toxicity in mice
Miho Watanabe-Takahashi; Shinji Yamasaki; Masayuki Murata; Fumi Kano; Jun Motoyama; Jyoji Yamate; Jumpei Omi; Waka Sato; Hirofumi Ukai; Kentaro Shimasaki; Masaya Ikegawa; Miwa Tamura-Nakano; Ryohei Yanoshita; Yuri Nishino; Atsuo Miyazawa; Yasuhiro Natori; Noriko Toyama-Sorimachi; Kiyotaka Nishikawa
Scientific Reports, 17 Jul. 2018, [Reviewed]
Abstract
Shiga toxin (Stx), a major virulence factor of enterohemorrhagic Escherichia coli (EHEC), is classified into two subgroups, Stx1 and Stx2. Clinical data clearly indicate that Stx2 is associated with more severe toxicity than Stx1, but the molecular mechanism underlying this difference is not fully understood. Here, we found that after being incorporated into target cells, Stx2, can be transported by recycling endosomes, as well as via the regular retrograde transport pathway. However, transport via recycling endosome did not occur with Stx1. We also found that Stx2 is actively released from cells in a receptor-recognizing B-subunit dependent manner. Part of the released Stx2 is associated with microvesicles, including exosome markers (referred to as exo-Stx2), whose origin is in the multivesicular bodies that formed from late/recycling endosomes. Finally, intravenous administration of exo-Stx2 to mice causes more lethality and tissue damage, especially severe renal dysfunction and tubular epithelial cell damage, compared to a free form of Stx2. Thus, the formation of exo-Stx2 might contribute to the severity of Stx2 in vivo, suggesting new therapeutic strategies against EHEC infections. - Contribution of the exosome-associated form of secreted endoplasmic reticulum aminopeptidase 1 to exosome-mediated macrophage activation.
Goto Y; Ogawa Y; Tsumoto H; Miura Y; Nakamura TJ; Ogawa K; Akimoto Y; Kawakami H; Endo T; Yanoshita R; Tsujimoto M
Biochimica et biophysica acta. Molecular cell research, Jun. 2018, [Reviewed] - マクロファージ活性化に関与するヒト唾液由来エキソソーム表面分子の解析
Mar. 2018 - ERAP1結合型エキソソームによるマクロファージの古典的活性化
Dec. 2017 - ヒト唾液由来エキソソームの表面に存在する分子は免疫活性化作用を制御する
Dec. 2017 - Hepatotoxicity, nephrotoxicity, and drug/chemical interaction toxicity of platinum nanoparticles in mice
Katsuhiro Isoda; T. Daibo; K. Yushina; Y. Yoshioka; Y. Tsutsumi; Y. Akimoto; H. Kawakami; Y. Taira; I. Taira; R. Yanoshita; T. Nishimura; I. Ishida
Pharmazie, 2017 - Characterization of membrane integrity and morphological stability of human salivary exosomes
Nahoko Kumeda; Yuko Ogawa; Yoshihiro Akimoto; Hayato Kawakami; Masafumi Tsujimoto; Ryohei Yanoshita
Biological and Pharmaceutical Bulletin, 2017, [Reviewed] - Next-Generation Sequencing of Protein-Coding and Long Non-protein-Coding RNAs in Two Types of Exosomes Derived from Human Whole Saliva.
Ogawa Y; Tsujimoto M; Yanoshita R
Biological & pharmaceutical bulletin, 2016, [Reviewed] - ヒト唾液エキソソームによるマクロファージの活性化
Oct. 2014 - 唾液エキソソームによるRAW264.7細胞の活性化
Mar. 2013 - Small RNA transcriptomes of two types of exosomes in human whole saliva determined by next generation sequencing.
Ogawa Y; Taketomi Y; Murakami M; Tsujimoto M; Yanoshita R
Biological & pharmaceutical bulletin, 2013, [Reviewed] - Selective accumulation of a novel antimalarial rhodacyanine derivative, SSJ-127, in an organelle of Plasmodium berghei
Mayumi Ikegami-Kawai; Chika Arai; Yuko Ogawa; Ryohei Yanoshita; Masataka Ihara
BIOORGANIC & MEDICINAL CHEMISTRY, Nov. 2010, [Reviewed] - Molecular cloning and characterization of ecto-5'-nucleotidase from the venoms of Gloydius blomhoffi.
Ogawa Y; Murayama N; Yanoshita R
Toxicon : official journal of the International Society on Toxinology, Sep. 2009, [Reviewed] - Exosome-like vesicles with dipeptidyl peptidase IV in human saliva.
Ogawa Y; Kanai-Azuma M; Akimoto Y; Kawakami H; Yanoshita R
Biological & pharmaceutical bulletin, Jun. 2008, [Reviewed] - Exosome-like vesicles in Gloydius blomhoffii blomhoffii venom.
Ogawa Y; Kanai-Azuma M; Akimoto Y; Kawakami H; Yanoshita R
Toxicon : official journal of the International Society on Toxinology, May 2008, [Reviewed] - Structural analysis of the interaction between Shiga toxin B subunits and linear polymers bearing clustered globotriose residues
M Watanabe; K Igai; K Matsuoka; A Miyagawa; T Watanabe; R Yanoshita; Y Samejima; D Terunuma; Y Natori; K Nishikawa
INFECTION AND IMMUNITY, Mar. 2006, [Reviewed] - Oral therapeutic agents with highly clustered globotriose for treatment of Shiga toxigenic Escherichia coli infections
M Watanabe; K Matsuoka; E Kita; K Igai; N Higashi; A Miyagawa; T Watanabe; R Yanoshita; Y Samejima; D Terunuma; Y Natori; K Nishikawa
JOURNAL OF INFECTIOUS DISEASES, Feb. 2004, [Reviewed] - Activated mast cells release extracellular type platelet-activating factor acetylhydrolase that contributes to autocrine inactivation of platelet-activating factor
K Nakajima; M Murakami; R Yanoshita; Y Samejima; K Karasawa; M Setaka; S Nojima; Kudo, I
JOURNAL OF BIOLOGICAL CHEMISTRY, Aug. 1997, [Reviewed] - GUINEA-PIG BONE-MARROW CELLS TREATED WITH PLATELET-ACTIVATING-FACTOR GENERATE FACTOR(S) WHICH AFFECTS THEIR DNA-SYNTHESIS AND MICROBICIDAL ACTIVITY
KUDO, I; T KATO; H HAYASHI; R YANOSHITA; K IKIZAWA; H UDA; K INOUE
LIPIDS, Dec. 1991, [Reviewed] - ANTITUMOR-ACTIVITY OF SYNTHETIC ALKYLPHOSPHOLIPIDS WITH OR WITHOUT PAF ACTIVITY
KUDO, I; S NOJIMA; HW CHANG; R YANOSHITA; H HAYASHI; E KONDO; H NOMURA; K INOUE
LIPIDS, Nov. 1987, [Reviewed]
MISC
- LPS-binding proteins in human salivary extracellular vesicles regulate the activation of macrophages.
小川裕子; 三浦ゆり; 後藤芳邦; 池本守; 秋元義弘; 遠藤玉夫; 矢ノ下良平
日本薬学会年会要旨集(Web), 2023 - アミノペプチダーゼに着目したヒト唾液由来細胞外小胞の性状解析
Mar. 2022 - Specification of extracellular vesicles characterized by their membrane proteins, APN/mucin1 or DPP IV/CD9
小川裕子; 三浦ゆり; 大西敦; 後藤芳邦; 青木一真; 池本守; 本車田悠希; 堤周平; 長島茉央; 廣谷莉花; 武井亮太朗; 秋元義弘; 遠藤玉夫; 矢ノ下良平
日本唾液腺学会誌, 2022 - Proteomic Analysis of Two Types of Exosomes in Human Whole Saliva
Yuko Ogawa; Yuri Miura; Akira Harazono; Masami Kanai-Azuma; Yoshihiro Akimoto; Hayato Kawakami; Teruhide Yamaguchi; Tosifusa Toda; Tamao Endo; Masayoshi Tsubuki; Ryohei Yanoshita
BIOLOGICAL & PHARMACEUTICAL BULLETIN, Jan. 2011 - ヘビ毒の膜結合性酵素の性状解析とcDNAクローニング
小川 裕子; 村山 信浩; 矢ノ下 良平
日本薬学会年会要旨集, Mar. 2008
(公社)日本薬学会 - タンビマムシ毒に含まれるアミノペプチダーゼの性状解析
小川 裕子; 矢ノ下 良平; 村山 信浩
日本薬学会年会要旨集, Mar. 2006
(公社)日本薬学会 - 2種のアマガサ蛇の毒に含まれるβ-ブンガロトキシンのcDNAクローニング
小川 裕子; 村山 信浩; 佐口 健一; 矢ノ下 良平; 佐藤 保; 樋口 成定; 鮫島 勇次
日本薬学会年会要旨集, Mar. 2004
(公社)日本薬学会 - 中国マムシ毒由来ジペプチジルアミノペプチダーゼIVの性状解析とcDNAクローニング
小川 裕子; 作田 朋子; 増井 由佳; 矢ノ下 良平; 村山 信浩; 樋口 成定; 鮫島 勇次
日本薬学会年会要旨集, Mar. 2004
(公社)日本薬学会 - cDNA cloning of bradykinin-potentiating peptides-C-type natriuretic peptide precursor, and characterization of the novel peptide Leu3-blomhotin from the venom of Agkistrodon blomhoffi
N Murayama; GH Michel; R Yanoshita; Y Samejima; K Saguchi; H Ohi; Y Fujita; S Higuchi
EUROPEAN JOURNAL OF BIOCHEMISTRY, Jul. 2000 - Blomhotin: a novel peptide with smooth muscle contractile activity identified in the venom of Agkistrodon halys blomhoffii
R Yanoshita; A Kasuga; S Inoue; K Ikeda; Y Samejima
TOXICON, Dec. 1999 - Inhibition of LysoPAF acetyltransferase activity by components of licorice root
S Nagumo; A Fukuju; M Takayama; H Nagai; R Yanoshita; Y Samejima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, Oct. 1999 - ブラジキニン増強ペプチド,平滑筋収縮ペプチド及びC型ナトリウム利尿ペプチドを一つの前駆体としてコードするニホンマムシ毒腺由来cDNAの解析と平滑筋収縮ペプチドの生物活性
村山 信浩; ミシェル・ジル; 大井 浩明; 藤田 吉明; 佐口 健一; 樋口 成定; 矢ノ下 良平; 鮫島 勇次
日本薬学会年会要旨集, Mar. 1999
(公社)日本薬学会 - PREFERENTIAL HYDROLYSIS OF PHOSPHATIDYLETHANOLAMINE IN RAT ISCHEMIC HEART HOMOGENATES DURING IN-VITRO INCUBATION
R KIKUCHIYANOSHITA; R YANOSHITA; KUDO, I; H ARAI; T TAKAMURA; K NOMOTO; K INOUE
JOURNAL OF BIOCHEMISTRY, Jul. 1993 - PAFの薬理作用(共著)
1992 - HYDROLYSIS OF PLATELET ACTIVATING FACTOR AND ITS METHYLATED ANALOGS BY ACETYLHYDROLASES
R YANOSHITA; KUDO, I; K IKIZAWA; HW CHANG; S KOBAYASHI; M OHNO; S NOJIMA; K INOUE
JOURNAL OF BIOCHEMISTRY, May 1988
Research Themes
- Mechanism of mucosal immunity activation by exosome
Grant-in-Aid for Scientific Research (C)
Teikyo Heisei University
21 Oct. 2016 - 31 Mar. 2019
Human whole saliva contains exosomes that have mucosal immunomodulatory potential. Immune-related proteins such as DPP IV and IgA, salivary proteins such as mucin 5B, and lipopolysaccharide (LPS) derived from oral bacteria were associated with surface of the exosomes.
Mucin 5B was digested with gastrointestinal enzymes. Although IgA and LPS were dissociated by size-exclusion chromatography, a part of them was tightly associated with the exosomes. Salivary exosome without the surface molecules increased production of nitric oxide (NO) from murine macrophages. In addition, we found that DPP IV may contribute to the NO production by collaboration with LPS. Salivary exosome is presumed to suppress the excessive activation in oral cavity and cause the activation of mucosal immunity after exposure to the gastrointestinal condition. - Molecular mechanism and regulation of immune activation cascade by aminopeptidase contained exosome.
Grant-in-Aid for Scientific Research (C)
Teikyo Heisei University
01 Apr. 2013 - 31 Mar. 2016
Human whole saliva contains two types of extracellular vesicles. One type highly expresses dipeptidyl peptidase-4 (DPP4) and the other type rarely expresses them. We designate former as DPP IV-exosome (DPP4exo), and later as deficient DPP IV-exosome (dDPP4exo). DPP4exo contained lipopolysaccharide (LPS) abundantly, and dDPP4exo contained it slightly. Although large amount of LPS interacted weakly with DPP4exo or existed separately forming micelle, part of LPS bind DPP4exo tightly. DPP4exo alone did not activate murine macrophage, however, DPP4exo with interferon-gamma produce high level of nitric oxide (NO) from murine macrophage. dDPP4exo did not cause NO production, even when interferon-gamma was added. DPP4exo is presumed to induce NO production from macrophage, when inflammation caused by bacterial infection, and associated with increased levels of interferon-gamma in oral cavity. - exosome