Detection of tumor ALK status in neuroblastoma patients using peripheral blood
New protocols based on ALK‐targeted therapy by crizotinib or other ALK‐targeting molecules have opened for the treatment of patients with neuroblastoma (NB) if their tumors showed mutation and/or amplification of the ALK gene. However, tumor samples are not always available for analysis of ALK mutat...
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| Veröffentlicht in: | Cancer medicine (Malden, MA) MA), 2015-04, Vol.4 (4), p.540-550 |
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| creator | Combaret, Valérie Iacono, Isabelle Bellini, Angela Bréjon, Stéphanie Bernard, Virginie Marabelle, Aurélien Coze, Carole Pierron, Gaelle Lapouble, Eve Schleiermacher, Gudrun Blay, Jean Yves |
| description | New protocols based on ALK‐targeted therapy by crizotinib or other ALK‐targeting molecules have opened for the treatment of patients with neuroblastoma (NB) if their tumors showed mutation and/or amplification of the ALK gene. However, tumor samples are not always available for analysis of ALK mutational status in particular at relapse. Here, we evaluated the ALK mutational status of NB samples by analysis of circulating DNA, using the droplet digital PCR (ddPCR) system. ddPCR assays was developed for the detection of ALK mutations at F1174 and R1275 hotspots found in NB tumors and was applied for the analysis of circulating DNA obtained from 200 μL of serum or plasma samples collected from 114 patients with NB. The mutations F1174L (exon 23 position 3520, T>C and position 3522, C>A) and the mutation R1275Q (exon 25 position 3824, G>A) were detected in circulating DNA. The sensitivity of our test was 100%, 85%, and 92%, respectively, and the specificity was 100%, 91%, and 98%, respectively. In conclusion, the assay that we have developed offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.
In the present report, by targeting the F1174 and R1275 hotspots of ALK gene and using the droplet digital PCR system, we demonstrate that ALK mutations are detectable in cell‐free plasma DNA from neuroblastoma patients showing a high risk of relapse. This assay offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help the clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available. |
| doi_str_mv | 10.1002/cam4.414 |
| format | Article |
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In the present report, by targeting the F1174 and R1275 hotspots of ALK gene and using the droplet digital PCR system, we demonstrate that ALK mutations are detectable in cell‐free plasma DNA from neuroblastoma patients showing a high risk of relapse. This assay offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help the clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.</description><identifier>ISSN: 2045-7634</identifier><identifier>ISSN: 1715-894X</identifier><identifier>EISSN: 2045-7634</identifier><identifier>DOI: 10.1002/cam4.414</identifier><identifier>PMID: 25653133</identifier><language>eng</language><publisher>United States: John Wiley & Sons, Inc</publisher><subject>ALK mutation ; ALK protein ; Biopsy ; Cancer ; Cancer Research ; Case-Control Studies ; cell‐free DNA ; Child ; ddPCR ; Deoxyribonucleic acid ; DNA ; DNA Mutational Analysis ; DNA, Neoplasm - genetics ; Exons ; Humans ; Kinases ; Life Sciences ; Metastasis ; Mutation ; Mutation - genetics ; Neuroblastoma ; Neuroblastoma - genetics ; Patients ; Peripheral blood ; Plasma ; Polymerase Chain Reaction - methods ; Receptor Protein-Tyrosine Kinases - genetics ; Tumors</subject><ispartof>Cancer medicine (Malden, MA), 2015-04, Vol.4 (4), p.540-550</ispartof><rights>2015 The Authors. published by John Wiley & Sons Ltd.</rights><rights>2015 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.</rights><rights>2015. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><rights>2015 The Authors. published by John Wiley & Sons Ltd. 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4384-3728c2cf7350dc955e9b504d719b2913568b50fd665bca9cbecd45998ae5a7873</citedby><cites>FETCH-LOGICAL-c4384-3728c2cf7350dc955e9b504d719b2913568b50fd665bca9cbecd45998ae5a7873</cites><orcidid>0000-0001-7190-120X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4402069/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4402069/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,1417,11562,27924,27925,45574,45575,46052,46476,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25653133$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://amu.hal.science/hal-01216548$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Combaret, Valérie</creatorcontrib><creatorcontrib>Iacono, Isabelle</creatorcontrib><creatorcontrib>Bellini, Angela</creatorcontrib><creatorcontrib>Bréjon, Stéphanie</creatorcontrib><creatorcontrib>Bernard, Virginie</creatorcontrib><creatorcontrib>Marabelle, Aurélien</creatorcontrib><creatorcontrib>Coze, Carole</creatorcontrib><creatorcontrib>Pierron, Gaelle</creatorcontrib><creatorcontrib>Lapouble, Eve</creatorcontrib><creatorcontrib>Schleiermacher, Gudrun</creatorcontrib><creatorcontrib>Blay, Jean Yves</creatorcontrib><title>Detection of tumor ALK status in neuroblastoma patients using peripheral blood</title><title>Cancer medicine (Malden, MA)</title><addtitle>Cancer Med</addtitle><description>New protocols based on ALK‐targeted therapy by crizotinib or other ALK‐targeting molecules have opened for the treatment of patients with neuroblastoma (NB) if their tumors showed mutation and/or amplification of the ALK gene. However, tumor samples are not always available for analysis of ALK mutational status in particular at relapse. Here, we evaluated the ALK mutational status of NB samples by analysis of circulating DNA, using the droplet digital PCR (ddPCR) system. ddPCR assays was developed for the detection of ALK mutations at F1174 and R1275 hotspots found in NB tumors and was applied for the analysis of circulating DNA obtained from 200 μL of serum or plasma samples collected from 114 patients with NB. The mutations F1174L (exon 23 position 3520, T>C and position 3522, C>A) and the mutation R1275Q (exon 25 position 3824, G>A) were detected in circulating DNA. The sensitivity of our test was 100%, 85%, and 92%, respectively, and the specificity was 100%, 91%, and 98%, respectively. In conclusion, the assay that we have developed offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.
In the present report, by targeting the F1174 and R1275 hotspots of ALK gene and using the droplet digital PCR system, we demonstrate that ALK mutations are detectable in cell‐free plasma DNA from neuroblastoma patients showing a high risk of relapse. This assay offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help the clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.</description><subject>ALK mutation</subject><subject>ALK protein</subject><subject>Biopsy</subject><subject>Cancer</subject><subject>Cancer Research</subject><subject>Case-Control Studies</subject><subject>cell‐free DNA</subject><subject>Child</subject><subject>ddPCR</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Mutational Analysis</subject><subject>DNA, Neoplasm - genetics</subject><subject>Exons</subject><subject>Humans</subject><subject>Kinases</subject><subject>Life Sciences</subject><subject>Metastasis</subject><subject>Mutation</subject><subject>Mutation - genetics</subject><subject>Neuroblastoma</subject><subject>Neuroblastoma - genetics</subject><subject>Patients</subject><subject>Peripheral blood</subject><subject>Plasma</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Receptor Protein-Tyrosine Kinases - genetics</subject><subject>Tumors</subject><issn>2045-7634</issn><issn>1715-894X</issn><issn>2045-7634</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kU1PHCEch4lpo0ZN-gkMiZd6GOWd4dJks9badLWX9kwYhnExM8MUGBu_vWzW-paUC28PD3_4AfAJozOMEDm3ZmBnDLMdsE8Q45UUlH14Nd4DRyndodIkIkLiXbBHuOAUU7oPbi5cdjb7MMLQwTwPIcLF6gdM2eQ5QT_C0c0xNL1JOQwGTiZ7N-YE5-THWzi56Ke1i6aHTR9Cewg-dqZP7uipPwC_L7_-Wl5Vq5_fvi8Xq8oyWrOKSlJbYjtJOWqt4typhiPWSqwaojDloi7zrhWCN9Yo2zjbMq5UbRw3spb0AHzZeqe5GVxrS0mlBj1FP5j4oIPx-u3O6Nf6NtxrxhBBQhXB6VawfnfsarHSmzWECRac1fe4sJ-fLovhz-xS1oNP1vW9GV2Yk8ZCUioFU6KgJ-_QuzDHsXyFJkQhjBgl6EVoY0gpuu65Aoz0JlO9yVSXTAt6_Pqhz-C_BAtQbYG_vncP_xXp5eKabYSP0MSpVA</recordid><startdate>201504</startdate><enddate>201504</enddate><creator>Combaret, Valérie</creator><creator>Iacono, Isabelle</creator><creator>Bellini, Angela</creator><creator>Bréjon, Stéphanie</creator><creator>Bernard, Virginie</creator><creator>Marabelle, Aurélien</creator><creator>Coze, Carole</creator><creator>Pierron, Gaelle</creator><creator>Lapouble, Eve</creator><creator>Schleiermacher, Gudrun</creator><creator>Blay, Jean Yves</creator><general>John Wiley & Sons, Inc</general><general>BC Decker Inc</general><general>BlackWell Publishing Ltd</general><scope>24P</scope><scope>WIN</scope><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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-7190-120X</orcidid></search><sort><creationdate>201504</creationdate><title>Detection of tumor ALK status in neuroblastoma patients using peripheral blood</title><author>Combaret, Valérie ; Iacono, Isabelle ; Bellini, Angela ; Bréjon, Stéphanie ; Bernard, Virginie ; Marabelle, Aurélien ; Coze, Carole ; Pierron, Gaelle ; Lapouble, Eve ; Schleiermacher, Gudrun ; Blay, Jean Yves</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4384-3728c2cf7350dc955e9b504d719b2913568b50fd665bca9cbecd45998ae5a7873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>ALK mutation</topic><topic>ALK protein</topic><topic>Biopsy</topic><topic>Cancer</topic><topic>Cancer Research</topic><topic>Case-Control Studies</topic><topic>cell‐free DNA</topic><topic>Child</topic><topic>ddPCR</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA Mutational Analysis</topic><topic>DNA, Neoplasm - genetics</topic><topic>Exons</topic><topic>Humans</topic><topic>Kinases</topic><topic>Life Sciences</topic><topic>Metastasis</topic><topic>Mutation</topic><topic>Mutation - genetics</topic><topic>Neuroblastoma</topic><topic>Neuroblastoma - genetics</topic><topic>Patients</topic><topic>Peripheral blood</topic><topic>Plasma</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Receptor Protein-Tyrosine Kinases - genetics</topic><topic>Tumors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Combaret, Valérie</creatorcontrib><creatorcontrib>Iacono, Isabelle</creatorcontrib><creatorcontrib>Bellini, Angela</creatorcontrib><creatorcontrib>Bréjon, Stéphanie</creatorcontrib><creatorcontrib>Bernard, Virginie</creatorcontrib><creatorcontrib>Marabelle, Aurélien</creatorcontrib><creatorcontrib>Coze, Carole</creatorcontrib><creatorcontrib>Pierron, Gaelle</creatorcontrib><creatorcontrib>Lapouble, Eve</creatorcontrib><creatorcontrib>Schleiermacher, Gudrun</creatorcontrib><creatorcontrib>Blay, Jean Yves</creatorcontrib><collection>Wiley-Blackwell Open Access Titles</collection><collection>Wiley Free Content</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Cancer medicine (Malden, MA)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Combaret, Valérie</au><au>Iacono, Isabelle</au><au>Bellini, Angela</au><au>Bréjon, Stéphanie</au><au>Bernard, Virginie</au><au>Marabelle, Aurélien</au><au>Coze, Carole</au><au>Pierron, Gaelle</au><au>Lapouble, Eve</au><au>Schleiermacher, Gudrun</au><au>Blay, Jean Yves</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of tumor ALK status in neuroblastoma patients using peripheral blood</atitle><jtitle>Cancer medicine (Malden, MA)</jtitle><addtitle>Cancer Med</addtitle><date>2015-04</date><risdate>2015</risdate><volume>4</volume><issue>4</issue><spage>540</spage><epage>550</epage><pages>540-550</pages><issn>2045-7634</issn><issn>1715-894X</issn><eissn>2045-7634</eissn><abstract>New protocols based on ALK‐targeted therapy by crizotinib or other ALK‐targeting molecules have opened for the treatment of patients with neuroblastoma (NB) if their tumors showed mutation and/or amplification of the ALK gene. However, tumor samples are not always available for analysis of ALK mutational status in particular at relapse. Here, we evaluated the ALK mutational status of NB samples by analysis of circulating DNA, using the droplet digital PCR (ddPCR) system. ddPCR assays was developed for the detection of ALK mutations at F1174 and R1275 hotspots found in NB tumors and was applied for the analysis of circulating DNA obtained from 200 μL of serum or plasma samples collected from 114 patients with NB. The mutations F1174L (exon 23 position 3520, T>C and position 3522, C>A) and the mutation R1275Q (exon 25 position 3824, G>A) were detected in circulating DNA. The sensitivity of our test was 100%, 85%, and 92%, respectively, and the specificity was 100%, 91%, and 98%, respectively. In conclusion, the assay that we have developed offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.
In the present report, by targeting the F1174 and R1275 hotspots of ALK gene and using the droplet digital PCR system, we demonstrate that ALK mutations are detectable in cell‐free plasma DNA from neuroblastoma patients showing a high risk of relapse. This assay offers a reliable, noninvasive blood test to assess ALK mutational status at F1174 and R1275 hotspots and should help the clinicians to identify patients showing an ALK mutation in particular when no tumor tissue is available.</abstract><cop>United States</cop><pub>John Wiley & Sons, Inc</pub><pmid>25653133</pmid><doi>10.1002/cam4.414</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-7190-120X</orcidid><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Wiley-Blackwell Open Access Titles; EZB-FREE-00999 freely available EZB journals; Wiley Online Library All Journals; PubMed Central |
| subjects | ALK mutation ALK protein Biopsy Cancer Cancer Research Case-Control Studies cell‐free DNA Child ddPCR Deoxyribonucleic acid DNA DNA Mutational Analysis DNA, Neoplasm - genetics Exons Humans Kinases Life Sciences Metastasis Mutation Mutation - genetics Neuroblastoma Neuroblastoma - genetics Patients Peripheral blood Plasma Polymerase Chain Reaction - methods Receptor Protein-Tyrosine Kinases - genetics Tumors |
| title | Detection of tumor ALK status in neuroblastoma patients using peripheral blood |
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