Spatial exosome analysis using cellulose nanofiber sheets reveals the location heterogeneity of extracellular vesicles

Akira Yokoi (), Kosuke Yoshida, Hirotaka Koga, Masami Kitagawa, Yukari Nagao, Mikiko Iida, Shota Kawaguchi, Min Zhang, Jun Nakayama, Yusuke Yamamoto, Yoshinobu Baba, Hiroaki Kajiyama and Takao Yasui ()
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Akira Yokoi: Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku
Kosuke Yoshida: Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku
Hirotaka Koga: Japan Science and Technology Agency (JST), FOREST, 4-1-8 Honcho, Kawaguchi
Masami Kitagawa: Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku
Yukari Nagao: Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku
Mikiko Iida: Nagoya University, Furo-cho, Chikusa-ku
Shota Kawaguchi: Nagoya University, Furo-cho, Chikusa-ku
Min Zhang: Nagoya University, Furo-cho, Chikusa-ku
Jun Nakayama: National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku
Yusuke Yamamoto: National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku
Yoshinobu Baba: Nagoya University, Furo-cho, Chikusa-ku
Hiroaki Kajiyama: Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku
Takao Yasui: Nagoya University Institute for Advanced Research, Furo-cho, Chikusa-ku

Nature Communications, 2023, vol. 14, issue 1, 1-15

Abstract: Abstract Extracellular vesicles (EVs), including exosomes, are recognized as promising functional targets involved in disease mechanisms. However, the intravital heterogeneity of EVs remains unclear, and the general limitation for analyzing EVs is the need for a certain volume of biofluids. Here, we present cellulose nanofiber (CNF) sheets to resolve these issues. We show that CNF sheets capture and preserve EVs from ~10 μL of biofluid and enable the analysis of bioactive molecules inside EVs. By attaching CNF sheets to moistened organs, we collect EVs in trace amounts of ascites, which is sufficient to perform small RNA sequence analyses. In an ovarian cancer mouse model, we demonstrate that CNF sheets enable the detection of cancer-associated miRNAs from the very early phase when mice did not have apparent ascites, and that EVs from different locations have unique miRNA profiles. By performing CNF sheet analyses in patients, we identify further location-based differences in EV miRNA profiles, with profiles reflecting disease conditions. We conduct spatial exosome analyses using CNF sheets to reveal that ascites EVs from cancer patients exhibit location-dependent heterogeneity. This technique could provide insights into EV biology and suggests a clinical strategy contributing to cancer diagnosis, staging evaluation, and therapy planning.

Date: 2023
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DOI: 10.1038/s41467-023-42593-9

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