@article{bd764e0322ba4719a386ac5d4ff960e3,
title = "Isolation, profiling, and tracking of extracellular vesicle cargo in Caenorhabditis elegans",
abstract = "Extracellular vesicles (EVs) may mediate intercellular communication by carrying protein and RNA cargo. The composition, biology, and roles of EVs in physiology and pathology have been primarily studied in the context of biofluids and in cultured mammalian cells. The experimental tractability of C. elegans makes for a powerful in vivo animal system to identify and study EV cargo from its cellular source. We developed an innovative method to label, track, and profile EVs using genetically encoded, fluorescent-tagged EV cargo and conducted a large-scale isolation and proteomic profiling. Nucleic acid binding proteins (∼200) are overrepresented in our dataset. By integrating our EV proteomic dataset with single-cell transcriptomic data, we identified and validated ciliary EV cargo: CD9-like tetraspanin (TSP-6), ectonucleotide pyrophosphatase/phosphodiesterase (ENPP-1), minichromosome maintenance protein (MCM-3), and double-stranded RNA transporter SID-2. C. elegans EVs also harbor RNA, suggesting that EVs may play a role in extracellular RNA-based communication.",
keywords = "ENPP1, MCM3, PKD2, polycystin, SID-2, cilia, ectosome, exosome, extracellular vesicle, seminal fluid",
author = "Nikonorova, {Inna A.} and Juan Wang and Cope, {Alexander L.} and Tilton, {Peter E.} and Power, {Kaiden M.} and Walsh, {Jonathon D.} and Akella, {Jyothi S.} and Krauchunas, {Amber R.} and Premal Shah and Barr, {Maureen M.}",
note = "Funding Information: We are grateful to Haiyan Zheng and Peter Loebel for assistance with the mass spectrometry analysis; Gloria Androwski and Helen Ushakov for outstanding technical assistance; Julie Claycomb, Rachel Kaletsky, Coleen Murphy, Tetsuya Nakamura, Joel Rosenbaum, Piali Sengupta, and Christopher Ward for discussions; members of the Boston and UCSF cilia supergroups and Rutgers C. elegans community for thought-provoking questions; Christine Maric-Bilkan for continued support and sound advice; and the three anonymous reviewers for constructive criticism. We also thank WormBase (release WS281) and WormBook that were used daily during this project. The work was funded by grants from the National Institutes of Health (NIH) DK116606, DK059418, and NS120745 to M.M.B.; National Science Foundation (NSF) DBI 1936046, NIH R35 GM124976, and start-up funds from HGINJ to P.S.; NIH K12 GM093854 INSPIRE (IRACDA NJ/NY) postdoctoral fellowships to J.D.W. and A.L.C.; and Donald Stahlin Memorial Fund. Protein identification was performed in the Biological Mass Spectrometry Facility of Robert Wood Johnson Medical School at Rutgers University supported by the NIH instrumentation grants S10OD025140 and S10OD016400. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). The Center for C. elegans Anatomy and WormAtlas provided valuable anatomical and ultrastructural resources (R24OD010943). Conceptualization, I.A.N. J.W. and M.M.B.; methodology, I.A.N. J.W. J.D.W. A.L.C. P.E.T. and P.S.; investigation, I.A.N. J.W. K.M.P. J.S.A. and A.L.C.; visualization, I.A.N. J.W. K.M.P. J.S.A. and P.E.T.; funding acquisition, M.M.B. J.D.W. and P.S.; project administration, I.A.N. J.W. M.M.B. and P.S.; supervision, M.M.B. P.S. and J.W.; writing ? original draft, I.A.N.; writing ? review & editing, I.A.N. J.W. A.L.C. K.M.P. J.D.W. J.S.A. A.R.K. P.S. and M.M.B. The authors declare no competing interests. Funding Information: We are grateful to Haiyan Zheng and Peter Loebel for assistance with the mass spectrometry analysis; Gloria Androwski and Helen Ushakov for outstanding technical assistance; Julie Claycomb, Rachel Kaletsky, Coleen Murphy, Tetsuya Nakamura, Joel Rosenbaum, Piali Sengupta, and Christopher Ward for discussions; members of the Boston and UCSF cilia supergroups and Rutgers C. elegans community for thought-provoking questions; Christine Maric-Bilkan for continued support and sound advice; and the three anonymous reviewers for constructive criticism. We also thank WormBase (release WS281) and WormBook that were used daily during this project. The work was funded by grants from the National Institutes of Health (NIH) DK116606 , DK059418 , and NS120745 to M.M.B.; National Science Foundation ( NSF) DBI 1936046 , NIH R35 GM124976 , and start-up funds from HGINJ to P.S.; NIH K12 GM093854 INSPIRE ( IRACDA NJ/NY ) postdoctoral fellowships to J.D.W. and A.L.C.; and Donald Stahlin Memorial Fund . Protein identification was performed in the Biological Mass Spectrometry Facility of Robert Wood Johnson Medical School at Rutgers University supported by the NIH instrumentation grants S10OD025140 and S10OD016400. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs ( P40 OD010440 ). The Center for C. elegans Anatomy and WormAtlas provided valuable anatomical and ultrastructural resources ( R24OD010943 ). Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
month = may,
day = "9",
doi = "10.1016/j.cub.2022.03.005",
language = "English (US)",
volume = "32",
pages = "1924--1936.e6",
journal = "Current Biology",
issn = "0960-9822",
publisher = "Cell Press",
number = "9",
}