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Differentiating extracellular vesicles by environmental disturbance method

Research Project

Project/Area Number 17H06255
Research Category

Grant-in-Aid for Challenging Research (Pioneering)

Allocation TypeSingle-year Grants
Research Field Molecular and Genome biology and related fields
Research InstitutionJapanese Foundation for Cancer Research

Principal Investigator

Shiba Kiyotaka  公益財団法人がん研究会, がん研究所 蛋白創製研究部, 部長 (40196415)

Project Period (FY) 2017-06-30 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥25,090,000 (Direct Cost: ¥19,300,000、Indirect Cost: ¥5,790,000)
Fiscal Year 2019: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
Fiscal Year 2018: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2017: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Keywords細胞外小胞 / EV / 生体膜 / エクソソーム / 診断学 / 細胞間相互作用 / 膜小胞 / 細胞生物学 / 細胞外分泌小胞 / エキソソーム / 微粒子 / マイクロ小胞 / EVs / Exosomes / 差分化 / 分析科学 / 細胞・組織 / 生体分子 / バイオテクノロジー / ナノバイオ
Outline of Final Research Achievements

Extracellular vesicles (EVs) in body fluids constitute heterogenous populations, which mirror their diverse parental cells as well as distinct EV-generation pathways. Here, by prolonging ultracentrifugation time to 96 h and fractionating EVs both by floating up or spinning down directions, we allowed 111 EV protein markers from the whole saliva of three healthy volunteers to attain equilibrium. Interestingly, the determined buoyant densities of the markers drifted in a specimen-specific manner, and drift patterns differentiated EVs into at least two subclasses. One class carried classical exosomal markers, such as CD63 and CD81, and the other was characterized by the molecules involved in membrane remodeling or vesicle trafficking. Distinct patterns of density drift may represent the differences in generation pathways of EVs.

Academic Significance and Societal Importance of the Research Achievements

唾液が含む1,429種のタンパク質のサブクラス化の情報を利用した、各種疾患に対する唾液診断法の開発が、今後進められると期待できる。同定したタンパク質の中には、SLC44A4、PMSA、CEACAM8/CD66b、Serum amyloid A-1 protein (SAA1)といった、広く知られている診断マーカーが含まれており、これらのマーカータンパク質は、細胞外小胞の積荷として体液中の存在している可能性が極めて高く、細胞外小胞にウィンドウを絞ったサンドイッチELISAの開発などにおいても、本研究の結果が利用されていくものと期待できる。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (13 results)

All 2019 2018 2017

All Journal Article (6 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 5 results,  Open Access: 3 results) Presentation (7 results) (of which Int'l Joint Research: 4 results,  Invited: 2 results)

  • [Journal Article] Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine2019

    • Author(s)
      S. Matsumura, T. Minamisawa, K. Suga, H. Kishita, T. Akagi, T. Ichiki, Y. Ichikawa, K. Shiba
    • Journal Title

      J. Extracell. Vesicles

      Volume: 8 Issue: 1 Pages: 1579541-1579541

    • DOI

      10.1080/20013078.2019.1579541

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Preferential capture of EpCAM-expressing extracellular vesicles on solid surfaces coated with an aptamer-conjugated zwitterionic polymer.2018

    • Author(s)
      Mitsutaka Yoshida, Kazuhiro Hibino, Satoshi Yamamoto, Sachiko Matsumura, Yasutomo Yajima, and Kiyotaka Shiba
    • Journal Title

      Biotechnology and Bioengineering

      Volume: Mar;115(3) Issue: 3 Pages: 536-544

    • DOI

      10.1002/bit.26489

    • NAID

      120006795409

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Host cell prediction of exosomes using morphological features on solid surfaces analyzed by machine learning.2018

    • Author(s)
      Ito, K., Ogawa, Y., Yokota, K., Matsumura, S., Minamisawa, T., Suga, K., Shiba, K., Kimura, Y., Hirano-Iwata, A., Takamura, Y. & Ogino, T.
    • Journal Title

      J Phys Chem B

      Volume: - Issue: 23 Pages: 6224-6235

    • DOI

      10.1021/acs.jpcb.8b01646

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Isolation of extracellular vesicles in saliva using density gradient ultracentrifugation.2017

    • Author(s)
      Iwai, K., Yamamoto, S., Yoshida, M. & Shiba, K.
    • Journal Title

      Methods Mol Biol.

      Volume: 1660 Pages: 343-350

    • DOI

      10.1007/978-1-4939-7253-1_27

    • ISBN
      9781493972517, 9781493972531
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Programmable Bio-surfaces for Biomedical Applications.2017

    • Author(s)
      Shiba, K.
    • Journal Title

      Adv Exp Med Biol.

      Volume: 1030 Pages: 1-20

    • DOI

      10.1007/978-3-319-66095-0_1

    • ISBN
      9783319660943, 9783319660950
    • Related Report
      2017 Annual Research Report
  • [Journal Article] Encryption of agonistic motifs for TLR4 into artificial antigens augmented the maturation of antigen-presenting cells.2017

    • Author(s)
      Ito M, Hayashi K, Minamisawa T, Homma S, Koido S, Shiba K.
    • Journal Title

      PLoS One.

      Volume: 12 Issue: 11 Pages: e0188934-e0188934

    • DOI

      10.1371/journal.pone.0188934

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Exposed aminophospholipids enriched in a subtype of small extracellular vesicles from tumor cell lines2019

    • Author(s)
      Matsumura S, Minamisawa T, Suga K, Shiba K
    • Organizer
      ISEV2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Inter Cavity Entire Veislcle 12019

    • Author(s)
      Shiba K
    • Organizer
      ICEV-1
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Extracellular vesicles in the oral cavity2019

    • Author(s)
      Shiba K
    • Organizer
      APSEV 2019 INAUGURAL ANNUAL MEETING
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 唾液含有微粒子の特徴と微粒子由来のRNA抽出条件についての検討2018

    • Author(s)
      平賀智豊、南澤宝后子、松村幸子、芝清隆
    • Organizer
      第5回日本細胞外小胞学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ナノ粒子を利用したエクソソームサブタイプの簡便な検出法の開発2018

    • Author(s)
      松村幸子、南澤宝美后、市川雄貴、芝清隆
    • Organizer
      第5回日本細胞外小胞学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Capturing EpCAM-positive extracellular vesicles by programmable bio-surface.2017

    • Author(s)
      Yoshida, M., Hibino, K., Matsumura, S., Minamisawa, T., Iwai, K., Yamamoto, S. & Shiba, K,
    • Organizer
      International Society for Extracellular Vesicles 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ナノテクノロジーと液性診断2017

    • Author(s)
      芝 清隆
    • Organizer
      2017年度生命科学系学会合同年次大会
    • Related Report
      2017 Annual Research Report

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Published: 2017-07-21   Modified: 2021-02-19  

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