Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers
MicroRNAs circulating in the blood, stabilized by complexation with proteins and/or additionally by encapsulation in lipid vesicles, are currently being evaluated as biomarkers. The consequences of their differential association with lipids/vesicles for their stability and use as biomarkers are larg...
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| creator | Köberle, Verena Pleli, Thomas Schmithals, Christian Augusto Alonso, Eduardo Haupenthal, Jörg Bönig, Halvard Peveling-Oberhag, Jan Biondi, Ricardo M Zeuzem, Stefan Kronenberger, Bernd Waidmann, Oliver Piiper, Albrecht |
| description | MicroRNAs circulating in the blood, stabilized by complexation with proteins and/or additionally by encapsulation in lipid vesicles, are currently being evaluated as biomarkers. The consequences of their differential association with lipids/vesicles for their stability and use as biomarkers are largely unexplored and are subject of the present study.
The levels of a set of selected microRNAs were determined by quantitative reverse-transcription PCR after extraction from sera or vesicle- and non-vesicle fractions prepared from sera. The stability of these microRNAs after incubation with RNase A or RNase inhibitor, an inhibitor of RNase A family enzymes was studied.
The levels of microRNA-1 and microRNA-122, but not those of microRNA-16, microRNA-21 and microRNA-142-3p, declined significantly during a 5-h incubation of the sera. RNase inhibitor prevented the loss of microRNAs in serum as well as the degradation of microRNA-122, a microRNA not expressed in blood cells, in whole blood. Stabilization of microRNA-122 was also achieved by hemolysis. Prolonged incubation of the sera led to enrichment of vesicle-associated relative to non-vesicle-associated microRNAs. Vesicle-associated microRNAs were more resistant to RNase A treatment than the respective microRNAs not associated with vesicles.
Serum microRNAs showed differential stability upon prolonged incubation. RNase inhibitor might be useful to robustly preserve the pattern of cell-free circulating microRNAs. In the case of microRNAs not expressed in blood cells this can also be achieved by hemolysis. Vesicle-associated microRNAs appeared to be more stable than those not associated with vesicles, which might be useful to disclose additional biomarker properties of miRNAs. |
| doi_str_mv | 10.1371/journal.pone.0075184 |
| format | Article |
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The levels of a set of selected microRNAs were determined by quantitative reverse-transcription PCR after extraction from sera or vesicle- and non-vesicle fractions prepared from sera. The stability of these microRNAs after incubation with RNase A or RNase inhibitor, an inhibitor of RNase A family enzymes was studied.
The levels of microRNA-1 and microRNA-122, but not those of microRNA-16, microRNA-21 and microRNA-142-3p, declined significantly during a 5-h incubation of the sera. RNase inhibitor prevented the loss of microRNAs in serum as well as the degradation of microRNA-122, a microRNA not expressed in blood cells, in whole blood. Stabilization of microRNA-122 was also achieved by hemolysis. Prolonged incubation of the sera led to enrichment of vesicle-associated relative to non-vesicle-associated microRNAs. Vesicle-associated microRNAs were more resistant to RNase A treatment than the respective microRNAs not associated with vesicles.
Serum microRNAs showed differential stability upon prolonged incubation. RNase inhibitor might be useful to robustly preserve the pattern of cell-free circulating microRNAs. In the case of microRNAs not expressed in blood cells this can also be achieved by hemolysis. Vesicle-associated microRNAs appeared to be more stable than those not associated with vesicles, which might be useful to disclose additional biomarker properties of miRNAs.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0075184</identifier><identifier>PMID: 24073250</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Biological markers ; Biomarkers ; Biomarkers, Tumor - blood ; Blood ; Blood cells ; Blood circulation ; Enzyme Inhibitors - pharmacology ; Enzymes ; Erythrocytes - metabolism ; Healthy Volunteers ; Hemolysis ; Hepatitis ; Hospitals ; Humans ; Inhibitors ; Interferon ; Kinases ; Lipids ; Liver cancer ; Medicine ; MicroRNA ; MicroRNAs ; MicroRNAs - blood ; MicroRNAs - chemistry ; miRNA ; Plasma ; Proteins ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonuclease ; Ribonuclease, Pancreatic - antagonists & inhibitors ; Ribonucleic acid ; RNA ; RNA Stability ; Serum - metabolism ; Stability ; Studies ; Transcription (Genetics) ; Urine ; Vesicles</subject><ispartof>PloS one, 2013-09, Vol.8 (9), p.e75184-e75184</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Köberle et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Köberle et al 2013 Köberle et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c743t-3803e889ed7aade000be2b9ec06f1738a09d60460dd56d9b1bbe96a000e342ab3</citedby><cites>FETCH-LOGICAL-c743t-3803e889ed7aade000be2b9ec06f1738a09d60460dd56d9b1bbe96a000e342ab3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3779196/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3779196/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79569,79570</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24073250$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Köberle, Verena</creatorcontrib><creatorcontrib>Pleli, Thomas</creatorcontrib><creatorcontrib>Schmithals, Christian</creatorcontrib><creatorcontrib>Augusto Alonso, Eduardo</creatorcontrib><creatorcontrib>Haupenthal, Jörg</creatorcontrib><creatorcontrib>Bönig, Halvard</creatorcontrib><creatorcontrib>Peveling-Oberhag, Jan</creatorcontrib><creatorcontrib>Biondi, Ricardo M</creatorcontrib><creatorcontrib>Zeuzem, Stefan</creatorcontrib><creatorcontrib>Kronenberger, Bernd</creatorcontrib><creatorcontrib>Waidmann, Oliver</creatorcontrib><creatorcontrib>Piiper, Albrecht</creatorcontrib><title>Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>MicroRNAs circulating in the blood, stabilized by complexation with proteins and/or additionally by encapsulation in lipid vesicles, are currently being evaluated as biomarkers. The consequences of their differential association with lipids/vesicles for their stability and use as biomarkers are largely unexplored and are subject of the present study.
The levels of a set of selected microRNAs were determined by quantitative reverse-transcription PCR after extraction from sera or vesicle- and non-vesicle fractions prepared from sera. The stability of these microRNAs after incubation with RNase A or RNase inhibitor, an inhibitor of RNase A family enzymes was studied.
The levels of microRNA-1 and microRNA-122, but not those of microRNA-16, microRNA-21 and microRNA-142-3p, declined significantly during a 5-h incubation of the sera. RNase inhibitor prevented the loss of microRNAs in serum as well as the degradation of microRNA-122, a microRNA not expressed in blood cells, in whole blood. Stabilization of microRNA-122 was also achieved by hemolysis. Prolonged incubation of the sera led to enrichment of vesicle-associated relative to non-vesicle-associated microRNAs. Vesicle-associated microRNAs were more resistant to RNase A treatment than the respective microRNAs not associated with vesicles.
Serum microRNAs showed differential stability upon prolonged incubation. RNase inhibitor might be useful to robustly preserve the pattern of cell-free circulating microRNAs. In the case of microRNAs not expressed in blood cells this can also be achieved by hemolysis. Vesicle-associated microRNAs appeared to be more stable than those not associated with vesicles, which might be useful to disclose additional biomarker properties of miRNAs.</description><subject>Biological markers</subject><subject>Biomarkers</subject><subject>Biomarkers, Tumor - blood</subject><subject>Blood</subject><subject>Blood cells</subject><subject>Blood circulation</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Enzymes</subject><subject>Erythrocytes - metabolism</subject><subject>Healthy Volunteers</subject><subject>Hemolysis</subject><subject>Hepatitis</subject><subject>Hospitals</subject><subject>Humans</subject><subject>Inhibitors</subject><subject>Interferon</subject><subject>Kinases</subject><subject>Lipids</subject><subject>Liver cancer</subject><subject>Medicine</subject><subject>MicroRNA</subject><subject>MicroRNAs</subject><subject>MicroRNAs - blood</subject><subject>MicroRNAs - chemistry</subject><subject>miRNA</subject><subject>Plasma</subject><subject>Proteins</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Ribonuclease</subject><subject>Ribonuclease, Pancreatic - antagonists & inhibitors</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA Stability</subject><subject>Serum - metabolism</subject><subject>Stability</subject><subject>Studies</subject><subject>Transcription (Genetics)</subject><subject>Urine</subject><subject>Vesicles</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk21v0zAQxyMEYmPwDRBEQkLwosVPiR1eIFVjg0oTk8bDW8txLq07Ny62gxifHrfNpgbtBYqlWOff_X13vsuy5xhNMeX43cr1vlN2unEdTBHiBRbsQXaMK0omJUH04cH-KHsSwgqhgoqyfJwdEYY4JQU6zq4_mrYFD100yuYhqtpYE29y1-YarJ20HiDXxuveqmi6Rb422rurL7PwPjfrjTU6mV0X8tb5PC7B-LyPSeLPzpyrkNfGrZW_Bh-eZo9aZQM8G_4n2ffzs2-nnycXl5_mp7OLieaMxgkViIIQFTRcqQYQQjWQugKNyhZzKhSqmhKxEjVNUTZVjesaqlIlDigjqqYn2cu97sa6IIc6BYkZZYzQoigSMd8TjVMrufEmRXgjnTJyZ3B-IZWPRluQhGtNQBCCeMsYFyItxStOa65xhXHS-jDc1tdraHQqpVd2JDo-6cxSLtwvSTmvcFUmgTeDgHc_ewhRrk3YFl914Ppd3LzglSAsoa_-Qe_PbqAWKiVgutale_VWVM5SBliUmG_jnt5Dpa-B9MapqVqT7COHtyOHxET4HReqD0HOv179P3v5Y8y-PmCXoGxcBmf7XV-NQbYHUweG4KG9KzJGcjsTt9WQ25mQw0wktxeHD3TndDsE9C8PCAc8</recordid><startdate>20130920</startdate><enddate>20130920</enddate><creator>Köberle, Verena</creator><creator>Pleli, Thomas</creator><creator>Schmithals, Christian</creator><creator>Augusto Alonso, Eduardo</creator><creator>Haupenthal, Jörg</creator><creator>Bönig, Halvard</creator><creator>Peveling-Oberhag, Jan</creator><creator>Biondi, Ricardo M</creator><creator>Zeuzem, Stefan</creator><creator>Kronenberger, Bernd</creator><creator>Waidmann, Oliver</creator><creator>Piiper, Albrecht</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PHGZM</scope><scope>PHGZT</scope><scope>PIMPY</scope><scope>PJZUB</scope><scope>PKEHL</scope><scope>PPXIY</scope><scope>PQEST</scope><scope>PQGLB</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130920</creationdate><title>Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers</title><author>Köberle, Verena ; Pleli, Thomas ; Schmithals, Christian ; Augusto Alonso, Eduardo ; Haupenthal, Jörg ; Bönig, Halvard ; Peveling-Oberhag, Jan ; Biondi, Ricardo M ; Zeuzem, Stefan ; Kronenberger, Bernd ; Waidmann, Oliver ; Piiper, Albrecht</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c743t-3803e889ed7aade000be2b9ec06f1738a09d60460dd56d9b1bbe96a000e342ab3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Biological markers</topic><topic>Biomarkers</topic><topic>Biomarkers, Tumor - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Köberle, Verena</au><au>Pleli, Thomas</au><au>Schmithals, Christian</au><au>Augusto Alonso, Eduardo</au><au>Haupenthal, Jörg</au><au>Bönig, Halvard</au><au>Peveling-Oberhag, Jan</au><au>Biondi, Ricardo M</au><au>Zeuzem, Stefan</au><au>Kronenberger, Bernd</au><au>Waidmann, Oliver</au><au>Piiper, Albrecht</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-09-20</date><risdate>2013</risdate><volume>8</volume><issue>9</issue><spage>e75184</spage><epage>e75184</epage><pages>e75184-e75184</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>MicroRNAs circulating in the blood, stabilized by complexation with proteins and/or additionally by encapsulation in lipid vesicles, are currently being evaluated as biomarkers. The consequences of their differential association with lipids/vesicles for their stability and use as biomarkers are largely unexplored and are subject of the present study.
The levels of a set of selected microRNAs were determined by quantitative reverse-transcription PCR after extraction from sera or vesicle- and non-vesicle fractions prepared from sera. The stability of these microRNAs after incubation with RNase A or RNase inhibitor, an inhibitor of RNase A family enzymes was studied.
The levels of microRNA-1 and microRNA-122, but not those of microRNA-16, microRNA-21 and microRNA-142-3p, declined significantly during a 5-h incubation of the sera. RNase inhibitor prevented the loss of microRNAs in serum as well as the degradation of microRNA-122, a microRNA not expressed in blood cells, in whole blood. Stabilization of microRNA-122 was also achieved by hemolysis. Prolonged incubation of the sera led to enrichment of vesicle-associated relative to non-vesicle-associated microRNAs. Vesicle-associated microRNAs were more resistant to RNase A treatment than the respective microRNAs not associated with vesicles.
Serum microRNAs showed differential stability upon prolonged incubation. RNase inhibitor might be useful to robustly preserve the pattern of cell-free circulating microRNAs. In the case of microRNAs not expressed in blood cells this can also be achieved by hemolysis. Vesicle-associated microRNAs appeared to be more stable than those not associated with vesicles, which might be useful to disclose additional biomarker properties of miRNAs.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24073250</pmid><doi>10.1371/journal.pone.0075184</doi><tpages>e75184</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2013-09, Vol.8 (9), p.e75184-e75184 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1434423555 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Biological markers Biomarkers Biomarkers, Tumor - blood Blood Blood cells Blood circulation Enzyme Inhibitors - pharmacology Enzymes Erythrocytes - metabolism Healthy Volunteers Hemolysis Hepatitis Hospitals Humans Inhibitors Interferon Kinases Lipids Liver cancer Medicine MicroRNA MicroRNAs MicroRNAs - blood MicroRNAs - chemistry miRNA Plasma Proteins Reverse Transcriptase Polymerase Chain Reaction Ribonuclease Ribonuclease, Pancreatic - antagonists & inhibitors Ribonucleic acid RNA RNA Stability Serum - metabolism Stability Studies Transcription (Genetics) Urine Vesicles |
| title | Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-20T07%3A14%3A17IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Differential%20stability%20of%20cell-free%20circulating%20microRNAs:%20implications%20for%20their%20utilization%20as%20biomarkers&rft.jtitle=PloS%20one&rft.au=K%C3%B6berle,%20Verena&rft.date=2013-09-20&rft.volume=8&rft.issue=9&rft.spage=e75184&rft.epage=e75184&rft.pages=e75184-e75184&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0075184&rft_dat=%3Cgale_plos_%3EA478186171%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1434423555&rft_id=info:pmid/24073250&rft_galeid=A478186171&rft_doaj_id=oai_doaj_org_article_27cc2e82207f44788788a7973b7c1911&rfr_iscdi=true |