Absolute Quantification of Dynamic Cellular Uptake of Small Extracellular Vesicles via Lanthanide Element Labeling and ICP–MS
Small extracellular vesicles (sEVs) are increasingly reported to play important roles in numerous physiological and pathological processes. Cellular uptake of sEVs is of great significance for functional regulation in recipient cells. Although various sEV quantification, labeling, and tracking metho...
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Veröffentlicht in: | Analytical chemistry (Washington) 2023-08, Vol.95 (32), p.11934-11942 |
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creator | Yang, Ningli Zhao, Chuanping Kong, Linlin Zhang, Baoying Han, Chunguang Zhang, Yangjun Qian, Xiaohong Qin, Weijie |
description | Small extracellular vesicles (sEVs) are increasingly reported to play important roles in numerous physiological and pathological processes. Cellular uptake of sEVs is of great significance for functional regulation in recipient cells. Although various sEV quantification, labeling, and tracking methods have been reported, it is still highly challenging to quantify the absolute amount of cellular uptake of sEVs and correlate this information with phenotypic variations in the recipient cell. Therefore, we developed a novel strategy using lanthanide element labeling and inductively coupled plasma–mass spectrometry (ICP–MS) for the absolute and sensitive quantification of sEVs. This strategy utilizes the chelation interaction between Eu3+ and the phosphate groups on the sEV membrane for specific labeling. sEVs internalized by cells can then be quantified by ICP–MS using a previously established linear relationship between the europium content and the particle numbers. High Eu labeling efficiency and stability were demonstrated by various evaluations, and no structural or functional alterations in the sEVs were discovered after Eu labeling. Application of this method revealed that 4020 ± 171 sEV particles/cell were internalized by HeLa cells at 37 °C and 61% uptake inhibition at 4 °C. Further investigation led to the quantitative differential analysis of sEV cellular uptake under the treatment of several chemical endocytosis inhibitors. A 23% strong inhibition indicated that HeLa cells uptake sEVs mainly through the macropinocytosis pathway. This facile labeling and absolute quantification strategy of sEVs with ppb-level high sensitivity is expected to become a potential tool for studying the functions of sEVs in intracellular communication and cargo transportation. |
doi_str_mv | 10.1021/acs.analchem.3c01421 |
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Cellular uptake of sEVs is of great significance for functional regulation in recipient cells. Although various sEV quantification, labeling, and tracking methods have been reported, it is still highly challenging to quantify the absolute amount of cellular uptake of sEVs and correlate this information with phenotypic variations in the recipient cell. Therefore, we developed a novel strategy using lanthanide element labeling and inductively coupled plasma–mass spectrometry (ICP–MS) for the absolute and sensitive quantification of sEVs. This strategy utilizes the chelation interaction between Eu3+ and the phosphate groups on the sEV membrane for specific labeling. sEVs internalized by cells can then be quantified by ICP–MS using a previously established linear relationship between the europium content and the particle numbers. High Eu labeling efficiency and stability were demonstrated by various evaluations, and no structural or functional alterations in the sEVs were discovered after Eu labeling. Application of this method revealed that 4020 ± 171 sEV particles/cell were internalized by HeLa cells at 37 °C and 61% uptake inhibition at 4 °C. Further investigation led to the quantitative differential analysis of sEV cellular uptake under the treatment of several chemical endocytosis inhibitors. A 23% strong inhibition indicated that HeLa cells uptake sEVs mainly through the macropinocytosis pathway. This facile labeling and absolute quantification strategy of sEVs with ppb-level high sensitivity is expected to become a potential tool for studying the functions of sEVs in intracellular communication and cargo transportation.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.3c01421</identifier><identifier>PMID: 37527423</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Analytical chemistry ; Chelation ; Chemistry ; Endocytosis ; Europium ; Extracellular vesicles ; Inductively coupled plasma mass spectrometry ; Intracellular signalling ; Labeling ; Mass spectrometry ; Mass spectroscopy ; Phenotypic variations ; Stability analysis ; Structure-function relationships ; Vesicles</subject><ispartof>Analytical chemistry (Washington), 2023-08, Vol.95 (32), p.11934-11942</ispartof><rights>2023 American Chemical Society</rights><rights>Copyright American Chemical Society Aug 15, 2023</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-a325t-4b91958e9d8e3bd4391a2a689b8930ea3ee6d920c1795d494f131ddeae2cc1383</cites><orcidid>0000-0002-7633-9786 ; 0000-0001-6245-9691</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.3c01421$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.3c01421$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>315,782,786,2767,27083,27931,27932,56745,56795</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37527423$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yang, Ningli</creatorcontrib><creatorcontrib>Zhao, Chuanping</creatorcontrib><creatorcontrib>Kong, Linlin</creatorcontrib><creatorcontrib>Zhang, Baoying</creatorcontrib><creatorcontrib>Han, Chunguang</creatorcontrib><creatorcontrib>Zhang, Yangjun</creatorcontrib><creatorcontrib>Qian, Xiaohong</creatorcontrib><creatorcontrib>Qin, Weijie</creatorcontrib><title>Absolute Quantification of Dynamic Cellular Uptake of Small Extracellular Vesicles via Lanthanide Element Labeling and ICP–MS</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Small extracellular vesicles (sEVs) are increasingly reported to play important roles in numerous physiological and pathological processes. Cellular uptake of sEVs is of great significance for functional regulation in recipient cells. Although various sEV quantification, labeling, and tracking methods have been reported, it is still highly challenging to quantify the absolute amount of cellular uptake of sEVs and correlate this information with phenotypic variations in the recipient cell. Therefore, we developed a novel strategy using lanthanide element labeling and inductively coupled plasma–mass spectrometry (ICP–MS) for the absolute and sensitive quantification of sEVs. This strategy utilizes the chelation interaction between Eu3+ and the phosphate groups on the sEV membrane for specific labeling. sEVs internalized by cells can then be quantified by ICP–MS using a previously established linear relationship between the europium content and the particle numbers. High Eu labeling efficiency and stability were demonstrated by various evaluations, and no structural or functional alterations in the sEVs were discovered after Eu labeling. Application of this method revealed that 4020 ± 171 sEV particles/cell were internalized by HeLa cells at 37 °C and 61% uptake inhibition at 4 °C. Further investigation led to the quantitative differential analysis of sEV cellular uptake under the treatment of several chemical endocytosis inhibitors. A 23% strong inhibition indicated that HeLa cells uptake sEVs mainly through the macropinocytosis pathway. 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Zhao, Chuanping ; Kong, Linlin ; Zhang, Baoying ; Han, Chunguang ; Zhang, Yangjun ; Qian, Xiaohong ; Qin, Weijie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a325t-4b91958e9d8e3bd4391a2a689b8930ea3ee6d920c1795d494f131ddeae2cc1383</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Analytical chemistry</topic><topic>Chelation</topic><topic>Chemistry</topic><topic>Endocytosis</topic><topic>Europium</topic><topic>Extracellular vesicles</topic><topic>Inductively coupled plasma mass spectrometry</topic><topic>Intracellular signalling</topic><topic>Labeling</topic><topic>Mass spectrometry</topic><topic>Mass spectroscopy</topic><topic>Phenotypic variations</topic><topic>Stability analysis</topic><topic>Structure-function relationships</topic><topic>Vesicles</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yang, Ningli</creatorcontrib><creatorcontrib>Zhao, Chuanping</creatorcontrib><creatorcontrib>Kong, Linlin</creatorcontrib><creatorcontrib>Zhang, Baoying</creatorcontrib><creatorcontrib>Han, Chunguang</creatorcontrib><creatorcontrib>Zhang, Yangjun</creatorcontrib><creatorcontrib>Qian, Xiaohong</creatorcontrib><creatorcontrib>Qin, Weijie</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yang, Ningli</au><au>Zhao, Chuanping</au><au>Kong, Linlin</au><au>Zhang, Baoying</au><au>Han, Chunguang</au><au>Zhang, Yangjun</au><au>Qian, Xiaohong</au><au>Qin, Weijie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Absolute Quantification of Dynamic Cellular Uptake of Small Extracellular Vesicles via Lanthanide Element Labeling and ICP–MS</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2023-08-15</date><risdate>2023</risdate><volume>95</volume><issue>32</issue><spage>11934</spage><epage>11942</epage><pages>11934-11942</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Small extracellular vesicles (sEVs) are increasingly reported to play important roles in numerous physiological and pathological processes. Cellular uptake of sEVs is of great significance for functional regulation in recipient cells. Although various sEV quantification, labeling, and tracking methods have been reported, it is still highly challenging to quantify the absolute amount of cellular uptake of sEVs and correlate this information with phenotypic variations in the recipient cell. Therefore, we developed a novel strategy using lanthanide element labeling and inductively coupled plasma–mass spectrometry (ICP–MS) for the absolute and sensitive quantification of sEVs. This strategy utilizes the chelation interaction between Eu3+ and the phosphate groups on the sEV membrane for specific labeling. sEVs internalized by cells can then be quantified by ICP–MS using a previously established linear relationship between the europium content and the particle numbers. High Eu labeling efficiency and stability were demonstrated by various evaluations, and no structural or functional alterations in the sEVs were discovered after Eu labeling. Application of this method revealed that 4020 ± 171 sEV particles/cell were internalized by HeLa cells at 37 °C and 61% uptake inhibition at 4 °C. Further investigation led to the quantitative differential analysis of sEV cellular uptake under the treatment of several chemical endocytosis inhibitors. A 23% strong inhibition indicated that HeLa cells uptake sEVs mainly through the macropinocytosis pathway. This facile labeling and absolute quantification strategy of sEVs with ppb-level high sensitivity is expected to become a potential tool for studying the functions of sEVs in intracellular communication and cargo transportation.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>37527423</pmid><doi>10.1021/acs.analchem.3c01421</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-7633-9786</orcidid><orcidid>https://orcid.org/0000-0001-6245-9691</orcidid></addata></record> |
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subjects | Analytical chemistry Chelation Chemistry Endocytosis Europium Extracellular vesicles Inductively coupled plasma mass spectrometry Intracellular signalling Labeling Mass spectrometry Mass spectroscopy Phenotypic variations Stability analysis Structure-function relationships Vesicles |
title | Absolute Quantification of Dynamic Cellular Uptake of Small Extracellular Vesicles via Lanthanide Element Labeling and ICP–MS |
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