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Visualization and tracking of tumour extracellular vesicle delivery and RNA translation using multiplexed reporters

Author

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  • Charles P. Lai

    (Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School)

  • Edward Y. Kim

    (Massachusetts General Hospital, and Program in Immunology, Harvard Medical School)

  • Christian E. Badr

    (Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School)

  • Ralph Weissleder

    (Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School)

  • Thorsten R. Mempel

    (Massachusetts General Hospital, and Program in Immunology, Harvard Medical School)

  • Bakhos A. Tannous

    (Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School)

  • Xandra O. Breakefield

    (Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School)

Abstract

Accurate spatiotemporal assessment of extracellular vesicle (EV) delivery and cargo RNA translation requires specific and robust live-cell imaging technologies. Here we engineer optical reporters to label multiple EV populations for visualization and tracking of tumour EV release, uptake and exchange between cell populations both in culture and in vivo. Enhanced green fluorescence protein (EGFP) and tandem dimer Tomato (tdTomato) were fused at NH2-termini with a palmitoylation signal (PalmGFP, PalmtdTomato) for EV membrane labelling. To monitor EV-RNA cargo, transcripts encoding PalmtdTomato were tagged with MS2 RNA binding sequences and detected by co-expression of bacteriophage MS2 coat protein fused with EGFP. By multiplexing fluorescent and bioluminescent EV membrane reporters, we reveal the rapid dynamics of both EV uptake and translation of EV-delivered cargo mRNAs in cancer cells that occurred within 1-hour post-horizontal transfer between cells. These studies confirm that EV-mediated communication is dynamic and multidirectional between cells with delivery of functional mRNA.

Suggested Citation

  • Charles P. Lai & Edward Y. Kim & Christian E. Badr & Ralph Weissleder & Thorsten R. Mempel & Bakhos A. Tannous & Xandra O. Breakefield, 2015. "Visualization and tracking of tumour extracellular vesicle delivery and RNA translation using multiplexed reporters," Nature Communications, Nature, vol. 6(1), pages 1-12, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8029
    DOI: 10.1038/ncomms8029
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    Cited by:

    1. Mokarram Hossain & Raymond Shim & Woo-Yong Lee & Arlene H. Sharpe & Paul Kubes, 2022. "Gata6+ resident peritoneal macrophages promote the growth of liver metastasis," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    2. Jinhong Xu & Le Cui & Jiaqi Wang & Shasha Zheng & Huahua Zhang & Shuo Ke & Xiaoqin Cao & Yanteng Shi & Jing Li & Ke Zen & Antonio Vidal-Puig & Chen-Yu Zhang & Liang Li & Xiaohong Jiang, 2023. "Cold-activated brown fat-derived extracellular vesicle-miR-378a-3p stimulates hepatic gluconeogenesis in male mice," Nature Communications, Nature, vol. 14(1), pages 1-19, December.

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