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エクソソームを介したトランスポゾン活性の発がんへの影響

Research Project

Project/Area Number 18J11869
Research Category

Grant-in-Aid for JSPS Fellows

Allocation TypeSingle-year Grants
Section国内
Research Field Cell biology
Research InstitutionUniversity of Tsukuba
Research Fellow 河村 有美  筑波大学, グローバル教育院, 特別研究員(DC2)
Project Period (FY) 2018-04-25 – 2020-03-31
Project Status Granted (Fiscal Year 2019)
Budget Amount *help
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2019: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2018: ¥800,000 (Direct Cost: ¥800,000)
Keywordsextracellular vesicle / retrotransposon / horizontal gene transfer
Outline of Annual Research Achievements

Long interspersed element-1 (LINE-1 or L1) retrotransposons replicate through a copy-and-paste mechanism using an RNA intermediate. However, little is known about the physical transmission of retrotransposon RNA between cells. To examine the horizontal transfer of an active human L1 retrotransposon mediated by extracellular vesicles (EVs), human cancer cells were transfected with an expression construct containing a retrotransposition-competent human L1 tagged with a reporter gene. Using this model, active retrotransposition events were detected by screening for the expression of the reporter gene inserted into the host genome by retrotransposition. EVs including exosomes and microvesicles were isolated from cells by differential centrifugation. The enrichment of L1-derived reporter RNA transcripts were detected in EVs isolated from cells expressing active L1 retrotransposition. The delivery of reporter RNA was confirmed in recipient cells, and reporter genes were detected in the genome of recipient cells. Additionally, employing qRT-PCR, we found that host-encoded factors are activated in response to increased exposure to L1-derived RNA transcripts in recipient cells. Our results suggest that the horizontal transfer of retrotransposons can occur through the incorporation of RNA intermediates delivered via EVs and may have important implications for the intercellular regulation of gene expression and gene function.

Current Status of Research Progress
Current Status of Research Progress

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

Reason

To examine the horizontal transfer of an active human L1 retrotransposon mediated by extracellular vesicles (EVs), human cancer cells were transfected with an expression construct containing a retrotransposition-competent human L1 tagged with a reporter gene. Using this model, active retrotransposition events were detected by screening for the expression of the reporter gene inserted into the host genome by retrotransposition. The enrichment of L1-derived reporter RNA transcripts were detected in EVs isolated from cells expressing active L1 retrotransposition. The delivery of reporter RNA was confirmed in recipient cells, and reporter genes were detected in the genome of recipient cells.

Strategy for Future Research Activity

EVs are packaged with abundant cellular information that has been shown to reflect the status of the cell, however, the mechanisms that regulate EV cargo selection is still largely unknown. Therefore, the biogenesis and cargo selection of EVs will be investigated to elucidate some of the biological processes that are regulated by EVs. The use of EVs as biomarkers for disease in liquid biopsy is a promising field, and overcoming the technical difficulties involved in the isolation and detection of EVs from biological samples would be beneficial in monitoring health and detecting diseases at an early stage. For this reason, new technologies in the isolation and detection of EVs from biological samles will be assessed, focusing on blood-derived EVs by microfluidics and the characterization of EV components.

Report

(1 results)
  • 2018 Annual Research Report

Research Products

(3 results)

All 2018

All Presentation (3 results) (of which Int'l Joint Research: 3 results)

  • [Presentation] Extracellular vesicles mediate the horizontal transfer of an active LINE-1 retrotransposon2018

    • Author(s)
      Yumi Kawamura
    • Organizer
      Annual Meeting of the International Society for Extracellular Vesicles
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Enrichment of long interspersed element-1 (LINE-1) RNA in extracellular vesicles from colorectal cancer patients2018

    • Author(s)
      Yumi Kawamura
    • Organizer
      The 2018 RNA Biology Symposium at the Cancer Science Institute of Singapore
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Hitchhiking extracellular vesicles to jump: Extracellular vesicles serve as delivery vehicles for LINE-1 retrotransposons2018

    • Author(s)
      Yumi Kawamura
    • Organizer
      Annual Meeting of the American Society for Exosomes and Microvesicles
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research

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Published: 2018-05-01   Modified: 2019-12-27  

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