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Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes. / Pakharukova, Maria Y; Savina, Ekaterina; Ponomarev, Dmitry V et al.

In: Journal of proteomics, Vol. 283-284, 104927, 15.07.2023.

Research output: Contribution to journalArticlepeer-review

Harvard

Pakharukova, MY, Savina, E, Ponomarev, DV, Gubanova, NV, Zaparina, O, Zakirova, EG, Cheng, G, Tikhonova, OV & Mordvinov, VA 2023, 'Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes', Journal of proteomics, vol. 283-284, 104927. https://doi.org/10.1016/j.jprot.2023.104927

APA

Pakharukova, M. Y., Savina, E., Ponomarev, D. V., Gubanova, N. V., Zaparina, O., Zakirova, E. G., Cheng, G., Tikhonova, O. V., & Mordvinov, V. A. (2023). Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes. Journal of proteomics, 283-284, [104927]. https://doi.org/10.1016/j.jprot.2023.104927

Vancouver

Pakharukova MY, Savina E, Ponomarev DV, Gubanova NV, Zaparina O, Zakirova EG et al. Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes. Journal of proteomics. 2023 Jul 15;283-284:104927. Epub 2023 May 22. doi: 10.1016/j.jprot.2023.104927

Author

Pakharukova, Maria Y ; Savina, Ekaterina ; Ponomarev, Dmitry V et al. / Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes. In: Journal of proteomics. 2023 ; Vol. 283-284.

BibTeX

@article{27ece8ef942f40d6bd3bb48d44fffb97,
title = "Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes",
abstract = "The epidemiologically important food-borne trematode Opisthorchis felineus infests the liver biliary tract of fish-eating mammals and causes disorders, including bile duct neoplasia. Many parasitic species release extracellular vesicles (EVs) that mediate host-parasite interaction. Currently, there is no information on O. felineus EVs. Using gel electrophoresis followed by liquid chromatography coupled with tandem mass spectrometry, we aimed to characterize the proteome of EVs released by the adult O. felineus liver fluke. Differential abundance of proteins between whole adult worms and EVs was assessed by semiquantitative iBAQ (intensity-based absolute quantification). Imaging, flow cytometry, inhibitor assays, and colocalization assays were performed to monitor the uptake of the EVs by H69 human cholangiocytes. The proteomic analysis reliably identified 168 proteins (at least two peptides matched a protein). Among major proteins of EVs were ferritin, tetraspanin CD63, helminth defense molecule 1, globin 3, saposin B type domain-containing protein, 60S ribosomal protein, glutathione S-transferase GST28, tubulin, and thioredoxin peroxidase. Moreover, as compared to the whole adult worm, EVs proved to be enriched with tetraspanin CD63, saposin B, helminth defense molecule 1, and Golgi-associated plant pathogenesis-related protein 1 (GAPR1). We showed that EVs are internalized by human H69 cholangiocytes via clathrin-dependent endocytosis, whereas phagocytosis and caveolin-dependent endocytosis do not play a substantial role in this process. Our study describes for the first time proteomes and differential abundance of proteins in whole adult O. felineus worms and EVs released by this food-borne trematode. Studies elucidating the regulatory role of individual components of EVs of liver flukes should be continued to determine which components of EV cargo play the most important part in the pathogenesis of fluke infection and in a closely linked pathology: bile duct neoplasia. SIGNIFICANCE: The food-borne trematode Opisthorchis felineus is a pathogen that causes hepatobiliary disorders in humans and animals. Our study describes for the first time the release of EVs by the liver fluke O. felineus, their microscopic and proteomic characterization, and internalization pathways by human cholangiocytes. Differential abundance of proteins between whole adult worms and EVs was assessed. EVs are enriched with canonical EV markers as well as parasite specific proteins, i.e. tetraspanin CD63, saposin B, helminth defense molecule 1, and others. Our findings will form the basis of the search for potential immunomodulatory candidates with therapeutic potential in the context of inflammatory diseases, as well as novel vaccine candidates.",
author = "Pakharukova, {Maria Y} and Ekaterina Savina and Ponomarev, {Dmitry V} and Gubanova, {Natalya V} and Oxana Zaparina and Zakirova, {Elvira G} and Guofeng Cheng and Tikhonova, {Olga V} and Mordvinov, {Viatcheslav A}",
note = "This work was supported by the Russian Science Foundation [grant number 22-24-20010 to M.Y.P.]. The funding agency had no role in this study, e.g., in study design, data collection, or decision to publish. Copyright {\textcopyright} 2023 Elsevier B.V. All rights reserved.",
year = "2023",
month = jul,
day = "15",
doi = "10.1016/j.jprot.2023.104927",
language = "English",
volume = "283-284",
journal = "Journal of proteomics",
issn = "1874-3919",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Proteomic characterization of Opisthorchis felineus exosome-like vesicles and their uptake by human cholangiocytes

AU - Pakharukova, Maria Y

AU - Savina, Ekaterina

AU - Ponomarev, Dmitry V

AU - Gubanova, Natalya V

AU - Zaparina, Oxana

AU - Zakirova, Elvira G

AU - Cheng, Guofeng

AU - Tikhonova, Olga V

AU - Mordvinov, Viatcheslav A

N1 - This work was supported by the Russian Science Foundation [grant number 22-24-20010 to M.Y.P.]. The funding agency had no role in this study, e.g., in study design, data collection, or decision to publish. Copyright © 2023 Elsevier B.V. All rights reserved.

PY - 2023/7/15

Y1 - 2023/7/15

N2 - The epidemiologically important food-borne trematode Opisthorchis felineus infests the liver biliary tract of fish-eating mammals and causes disorders, including bile duct neoplasia. Many parasitic species release extracellular vesicles (EVs) that mediate host-parasite interaction. Currently, there is no information on O. felineus EVs. Using gel electrophoresis followed by liquid chromatography coupled with tandem mass spectrometry, we aimed to characterize the proteome of EVs released by the adult O. felineus liver fluke. Differential abundance of proteins between whole adult worms and EVs was assessed by semiquantitative iBAQ (intensity-based absolute quantification). Imaging, flow cytometry, inhibitor assays, and colocalization assays were performed to monitor the uptake of the EVs by H69 human cholangiocytes. The proteomic analysis reliably identified 168 proteins (at least two peptides matched a protein). Among major proteins of EVs were ferritin, tetraspanin CD63, helminth defense molecule 1, globin 3, saposin B type domain-containing protein, 60S ribosomal protein, glutathione S-transferase GST28, tubulin, and thioredoxin peroxidase. Moreover, as compared to the whole adult worm, EVs proved to be enriched with tetraspanin CD63, saposin B, helminth defense molecule 1, and Golgi-associated plant pathogenesis-related protein 1 (GAPR1). We showed that EVs are internalized by human H69 cholangiocytes via clathrin-dependent endocytosis, whereas phagocytosis and caveolin-dependent endocytosis do not play a substantial role in this process. Our study describes for the first time proteomes and differential abundance of proteins in whole adult O. felineus worms and EVs released by this food-borne trematode. Studies elucidating the regulatory role of individual components of EVs of liver flukes should be continued to determine which components of EV cargo play the most important part in the pathogenesis of fluke infection and in a closely linked pathology: bile duct neoplasia. SIGNIFICANCE: The food-borne trematode Opisthorchis felineus is a pathogen that causes hepatobiliary disorders in humans and animals. Our study describes for the first time the release of EVs by the liver fluke O. felineus, their microscopic and proteomic characterization, and internalization pathways by human cholangiocytes. Differential abundance of proteins between whole adult worms and EVs was assessed. EVs are enriched with canonical EV markers as well as parasite specific proteins, i.e. tetraspanin CD63, saposin B, helminth defense molecule 1, and others. Our findings will form the basis of the search for potential immunomodulatory candidates with therapeutic potential in the context of inflammatory diseases, as well as novel vaccine candidates.

AB - The epidemiologically important food-borne trematode Opisthorchis felineus infests the liver biliary tract of fish-eating mammals and causes disorders, including bile duct neoplasia. Many parasitic species release extracellular vesicles (EVs) that mediate host-parasite interaction. Currently, there is no information on O. felineus EVs. Using gel electrophoresis followed by liquid chromatography coupled with tandem mass spectrometry, we aimed to characterize the proteome of EVs released by the adult O. felineus liver fluke. Differential abundance of proteins between whole adult worms and EVs was assessed by semiquantitative iBAQ (intensity-based absolute quantification). Imaging, flow cytometry, inhibitor assays, and colocalization assays were performed to monitor the uptake of the EVs by H69 human cholangiocytes. The proteomic analysis reliably identified 168 proteins (at least two peptides matched a protein). Among major proteins of EVs were ferritin, tetraspanin CD63, helminth defense molecule 1, globin 3, saposin B type domain-containing protein, 60S ribosomal protein, glutathione S-transferase GST28, tubulin, and thioredoxin peroxidase. Moreover, as compared to the whole adult worm, EVs proved to be enriched with tetraspanin CD63, saposin B, helminth defense molecule 1, and Golgi-associated plant pathogenesis-related protein 1 (GAPR1). We showed that EVs are internalized by human H69 cholangiocytes via clathrin-dependent endocytosis, whereas phagocytosis and caveolin-dependent endocytosis do not play a substantial role in this process. Our study describes for the first time proteomes and differential abundance of proteins in whole adult O. felineus worms and EVs released by this food-borne trematode. Studies elucidating the regulatory role of individual components of EVs of liver flukes should be continued to determine which components of EV cargo play the most important part in the pathogenesis of fluke infection and in a closely linked pathology: bile duct neoplasia. SIGNIFICANCE: The food-borne trematode Opisthorchis felineus is a pathogen that causes hepatobiliary disorders in humans and animals. Our study describes for the first time the release of EVs by the liver fluke O. felineus, their microscopic and proteomic characterization, and internalization pathways by human cholangiocytes. Differential abundance of proteins between whole adult worms and EVs was assessed. EVs are enriched with canonical EV markers as well as parasite specific proteins, i.e. tetraspanin CD63, saposin B, helminth defense molecule 1, and others. Our findings will form the basis of the search for potential immunomodulatory candidates with therapeutic potential in the context of inflammatory diseases, as well as novel vaccine candidates.

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85160448091&origin=inward&txGid=bd65393b3086eda7f980322abd0a29ad

U2 - 10.1016/j.jprot.2023.104927

DO - 10.1016/j.jprot.2023.104927

M3 - Article

C2 - 37225040

VL - 283-284

JO - Journal of proteomics

JF - Journal of proteomics

SN - 1874-3919

M1 - 104927

ER -

ID: 50152960