ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing

Background: ALK rearrangements are present in 5% of nonsmall cell lung cancer (NSCLC) tumors and identify patients who can benefit from ALK inhibitors. ALK fusions testing using liquid biopsies, although challenging, can expand the therapeutic options for ALK-positive NSCLC patients considerably. RN...

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Autores: Sánchez‑Herrero, Estela, Campos-Silva, Carmen, Cáceres-Martell, Yaiza, Robado de Lope, Lucía, Sanz-Moreno, Sandra, Serna-Blasco, Roberto, Rodríguez-Festa, Alejandro, Ares Trotta, Dunixe, Martín-Acosta, Paloma, Patino, Cristina, Coronado, María José, Beneítez-Martínez, Alexandra, Jara-Acevedo, Ricardo, Lago-Baameiro, Nerea, Camino, Tamara, Cruz-Bermúdez, Alberto, Pardo, María, González-Rumayor, Víctor, Valés-Gómez, Mar, Provencio, Mariano, Romero, Atocha
Formato: artículo
Fecha de publicación:2022
País:España
Recursos:Universidad Loyola Andalucía
Repositorio:Brújula
OAI Identifier:oai:repositorio.uloyola.es:20.500.12412/5677
Acesso em linha:https://hdl.handle.net/20.500.12412/5677
Access Level:acceso abierto
Palavra-chave:ALK-TKI
EML4-ALK
NSCLC
Extracellular vesicles
Liquid biopsy
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spelling ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive TestingSánchez‑Herrero, EstelaCampos-Silva, CarmenCáceres-Martell, YaizaRobado de Lope, LucíaSanz-Moreno, SandraSerna-Blasco, RobertoRodríguez-Festa, AlejandroAres Trotta, DunixeMartín-Acosta, PalomaPatino, CristinaCoronado, María JoséBeneítez-Martínez, AlexandraJara-Acevedo, RicardoLago-Baameiro, NereaCamino, TamaraCruz-Bermúdez, AlbertoPardo, MaríaGonzález-Rumayor, VíctorValés-Gómez, MarProvencio, MarianoRomero, AtochaALK-TKIEML4-ALKNSCLCExtracellular vesiclesLiquid biopsyBackground: ALK rearrangements are present in 5% of nonsmall cell lung cancer (NSCLC) tumors and identify patients who can benefit from ALK inhibitors. ALK fusions testing using liquid biopsies, although challenging, can expand the therapeutic options for ALK-positive NSCLC patients considerably. RNA inside extracellular vesicles (EVs) is protected from RNases and other environmental factors, constituting a promising source for noninvasive fusion transcript detection. Methods: EVs from H3122 and H2228 cell lines, harboring EML4-ALK variant 1 (E13; A20) and variant 3 (E6a/b; A20), respectively, were successfully isolated by sequential centrifugation of cell culture supernatants. EVs were also isolated from plasma samples of 16 ALK-positive NSCLC patients collected before treatment initiation. Results: Purified EVs from cell cultures were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and flow cytometry. Western blot and confocal microscopy confirmed the expression of EV-specific markers as well as the expression of EML4-ALK-fusion proteins in EV fractions from H3122 and H2228 cell lines. In addition, RNA from EV fractions derived from cell culture was analyzed by digital PCR (dPCR) and ALK-fusion transcripts were clearly detected. Similarly, plasma-derived EVs were characterized by NTA, flow cytometry, and the ExoView platform, the last showing that EV-specific markers captured EV populations containing ALK-fusion protein. Finally, ALK fusions were identified in 50% (8/16) of plasma EV-enriched fractions by dPCR, confirming the presence of fusion transcripts in EV fractions. Conclusions: ALK-fusion transcripts can be detected in EV-enriched fractions. These results set the stage for the development of EV-based noninvasive ALK testing.2022info:eu-repo/semantics/articlehttps://hdl.handle.net/20.500.12412/5677reponame:Brújulainstname:Universidad Loyola AndalucíaIngléshttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:repositorio.uloyola.es:20.500.12412/56772026-06-24T12:48:37Z
dc.title.none.fl_str_mv ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
title ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
spellingShingle ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
Sánchez‑Herrero, Estela
ALK-TKI
EML4-ALK
NSCLC
Extracellular vesicles
Liquid biopsy
title_short ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
title_full ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
title_fullStr ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
title_full_unstemmed ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
title_sort ALK-Fusion Transcripts Can Be Detected in Extracellular Vesicles (EVs) from Nonsmall Cell Lung Cancer Cell Lines and Patient Plasma: Toward EV-Based Noninvasive Testing
dc.creator.none.fl_str_mv Sánchez‑Herrero, Estela
Campos-Silva, Carmen
Cáceres-Martell, Yaiza
Robado de Lope, Lucía
Sanz-Moreno, Sandra
Serna-Blasco, Roberto
Rodríguez-Festa, Alejandro
Ares Trotta, Dunixe
Martín-Acosta, Paloma
Patino, Cristina
Coronado, María José
Beneítez-Martínez, Alexandra
Jara-Acevedo, Ricardo
Lago-Baameiro, Nerea
Camino, Tamara
Cruz-Bermúdez, Alberto
Pardo, María
González-Rumayor, Víctor
Valés-Gómez, Mar
Provencio, Mariano
Romero, Atocha
author Sánchez‑Herrero, Estela
author_facet Sánchez‑Herrero, Estela
Campos-Silva, Carmen
Cáceres-Martell, Yaiza
Robado de Lope, Lucía
Sanz-Moreno, Sandra
Serna-Blasco, Roberto
Rodríguez-Festa, Alejandro
Ares Trotta, Dunixe
Martín-Acosta, Paloma
Patino, Cristina
Coronado, María José
Beneítez-Martínez, Alexandra
Jara-Acevedo, Ricardo
Lago-Baameiro, Nerea
Camino, Tamara
Cruz-Bermúdez, Alberto
Pardo, María
González-Rumayor, Víctor
Valés-Gómez, Mar
Provencio, Mariano
Romero, Atocha
author_role author
author2 Campos-Silva, Carmen
Cáceres-Martell, Yaiza
Robado de Lope, Lucía
Sanz-Moreno, Sandra
Serna-Blasco, Roberto
Rodríguez-Festa, Alejandro
Ares Trotta, Dunixe
Martín-Acosta, Paloma
Patino, Cristina
Coronado, María José
Beneítez-Martínez, Alexandra
Jara-Acevedo, Ricardo
Lago-Baameiro, Nerea
Camino, Tamara
Cruz-Bermúdez, Alberto
Pardo, María
González-Rumayor, Víctor
Valés-Gómez, Mar
Provencio, Mariano
Romero, Atocha
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv ALK-TKI
EML4-ALK
NSCLC
Extracellular vesicles
Liquid biopsy
topic ALK-TKI
EML4-ALK
NSCLC
Extracellular vesicles
Liquid biopsy
description Background: ALK rearrangements are present in 5% of nonsmall cell lung cancer (NSCLC) tumors and identify patients who can benefit from ALK inhibitors. ALK fusions testing using liquid biopsies, although challenging, can expand the therapeutic options for ALK-positive NSCLC patients considerably. RNA inside extracellular vesicles (EVs) is protected from RNases and other environmental factors, constituting a promising source for noninvasive fusion transcript detection. Methods: EVs from H3122 and H2228 cell lines, harboring EML4-ALK variant 1 (E13; A20) and variant 3 (E6a/b; A20), respectively, were successfully isolated by sequential centrifugation of cell culture supernatants. EVs were also isolated from plasma samples of 16 ALK-positive NSCLC patients collected before treatment initiation. Results: Purified EVs from cell cultures were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and flow cytometry. Western blot and confocal microscopy confirmed the expression of EV-specific markers as well as the expression of EML4-ALK-fusion proteins in EV fractions from H3122 and H2228 cell lines. In addition, RNA from EV fractions derived from cell culture was analyzed by digital PCR (dPCR) and ALK-fusion transcripts were clearly detected. Similarly, plasma-derived EVs were characterized by NTA, flow cytometry, and the ExoView platform, the last showing that EV-specific markers captured EV populations containing ALK-fusion protein. Finally, ALK fusions were identified in 50% (8/16) of plasma EV-enriched fractions by dPCR, confirming the presence of fusion transcripts in EV fractions. Conclusions: ALK-fusion transcripts can be detected in EV-enriched fractions. These results set the stage for the development of EV-based noninvasive ALK testing.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.12412/5677
url https://hdl.handle.net/20.500.12412/5677
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv reponame:Brújula
instname:Universidad Loyola Andalucía
instname_str Universidad Loyola Andalucía
reponame_str Brújula
collection Brújula
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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