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CME Evaluation

Oral Communication Session

Venous Thromboembolism

Session: Cancer associated thrombosis - Mechanisms

OC 12.4 - Cancer-associated fibroblasts promote venous thrombosis by releasing podoplanin-positive extracellular vesicles.

Sunday, June 23, 2024
10:15 – 10:30 ICT
Room: Ballroom B2

    Presenting Author(s)

  • Toshiaki Shirai, PhD

    Assisitant Professor
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

    • Co-Author(s)

  • Nagaharu Tsukiji, PhD

    Senior Assistant Professor/Lecturer
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

  • TS

    Tomoyuki Sasaki

    Associate Professor
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

  • SO

    Saori Oishi

    Assisitant Professor
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

  • Ryohei Yokomori

    Ph.D student
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

  • KT

    Katsuhiro Takano

    Lecturer
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

  • KI

    Katsue Inoue

    Professor
    Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi
    Chuo, Yamanashi, Japan

Background: Podoplanin (PDPN), a sialo glycoprotein, induces platelet aggregation through C-type lectin-like receptor 2 (CLEC-2). Despite the promising connection between PDPN and CLEC-2 in cancer-associated thrombosis (CAT), PDPN isn't found on the surface of tumor cells in high-risk CAT, including pancreas and lung. Cancer-associated fibroblasts (CAFs) promotes cancer progression through cell―cell interactions and by releasing extracellular vesicles (EVs). Interestingly, PDPN is present in CAFs across different tumor types.

Aims: Our Study focused on determining if CAFs aggravate CAT via the PDPN/CLEC-2 pathway within the preclinical lung cancer model using Lewis Lung Carcinoma (LLC).

Methods: Cells expressing PDPN within the LLC tumor microenvironment was histologically analyzed. The presence of podoplanin in EVs from CAFs-conditioned medium and LLC tumor tissues was identified through immunoblotting. Mouse platelet activation (P-selectin and JON/A binding) and aggregation were analyzed by flow cytometry. As an acute venous thrombosis model, the femoral vein of mice injected with CAF-EVs or LLC tumor-bearing state was injured by 2.5% ferric chloride. To induce CLEC-2 depletion on platelets, mice were treated with rat anti-mouse CLEC-2 monoclonal antibody 2A2B10.

Results: CAFs expressing CD90, PDGFRβ, HSP47, CD34, and vimentin, co-expressed PDPN. EVs carried PDPN in both microvesicles and exosomes. EVs induced mouse platelet activation/aggregation in a CLEC-2-dependent manner. In vivo, mice injected with CAF-EVs and LLC tumor-bearing state showed a shorter occlusion time compared to the healthy control. CLEC-2-depleted mice extended occlusion time in the presence of CAF-EVs or LLC tumor but not in the healthy condition.

Conclusion(s): CAFs that express podoplanin release this glycoprotein through EVs, aggravating venous thrombosis.