Cargando…
EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity
EML4-ALK is an oncogenic fusion protein present in approximately 5% of non–small cell lung cancers (NSCLC). Alternative breakpoints in the gene encoding EML4 result in distinct variants that are linked to markedly different patient outcomes. Patients with EML4-ALK variant 3 (V3) respond poorly to AL...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Association for Cancer Research
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9381094/ https://www.ncbi.nlm.nih.gov/pubmed/35656694 http://dx.doi.org/10.1158/1541-7786.MCR-21-1010 |
_version_ | 1784769007358836736 |
---|---|
author | Lucken, Kellie O'Regan, Laura Choi, Jene Sampson, Josephina Pashley, Sarah L. Bayliss, Richard Khan, Sam Fry, Andrew M. |
author_facet | Lucken, Kellie O'Regan, Laura Choi, Jene Sampson, Josephina Pashley, Sarah L. Bayliss, Richard Khan, Sam Fry, Andrew M. |
author_sort | Lucken, Kellie |
collection | PubMed |
description | EML4-ALK is an oncogenic fusion protein present in approximately 5% of non–small cell lung cancers (NSCLC). Alternative breakpoints in the gene encoding EML4 result in distinct variants that are linked to markedly different patient outcomes. Patients with EML4-ALK variant 3 (V3) respond poorly to ALK inhibitors and have lower survival rates compared with patients with other common variants, such as V1. Here, we use isogenic Beas-2B bronchial epithelial cell lines expressing EML4-ALK V1 or V3, as well as ALK-positive NSCLC patient cells that express V1 (H3122 cells) or V3 (H2228 cells), to show that EML4-ALK V3 but not V1 leads to hyperstabilized K-fibers in mitosis, as well as errors in chromosome congression and segregation. This is consistent with our observation that EML4-ALK V3 but not V1 localizes to spindle microtubules and that wild-type EML4 is a microtubule stabilizing protein. In addition, cells expressing EML4-ALK V3 exhibit loss of spindle assembly checkpoint control that is at least in part dependent on ALK catalytic activity. Finally, we demonstrate that cells expressing EML4-ALK V3 have increased sensitivity to microtubule poisons that interfere with mitotic spindle assembly, whereas combination treatment with paclitaxel and clinically approved ALK inhibitors leads to a synergistic response in terms of reduced survival of H2228 cells. IMPLICATIONS: This study suggests that combining the microtubule poison, paclitaxel, with targeted ALK inhibitors may provide an effective new treatment option for patients with NSCLC with tumors that express the EML4-ALK V3 oncogenic fusion. |
format | Online Article Text |
id | pubmed-9381094 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Association for Cancer Research |
record_format | MEDLINE/PubMed |
spelling | pubmed-93810942022-10-31 EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity Lucken, Kellie O'Regan, Laura Choi, Jene Sampson, Josephina Pashley, Sarah L. Bayliss, Richard Khan, Sam Fry, Andrew M. Mol Cancer Res Cancer Genes and Networks EML4-ALK is an oncogenic fusion protein present in approximately 5% of non–small cell lung cancers (NSCLC). Alternative breakpoints in the gene encoding EML4 result in distinct variants that are linked to markedly different patient outcomes. Patients with EML4-ALK variant 3 (V3) respond poorly to ALK inhibitors and have lower survival rates compared with patients with other common variants, such as V1. Here, we use isogenic Beas-2B bronchial epithelial cell lines expressing EML4-ALK V1 or V3, as well as ALK-positive NSCLC patient cells that express V1 (H3122 cells) or V3 (H2228 cells), to show that EML4-ALK V3 but not V1 leads to hyperstabilized K-fibers in mitosis, as well as errors in chromosome congression and segregation. This is consistent with our observation that EML4-ALK V3 but not V1 localizes to spindle microtubules and that wild-type EML4 is a microtubule stabilizing protein. In addition, cells expressing EML4-ALK V3 exhibit loss of spindle assembly checkpoint control that is at least in part dependent on ALK catalytic activity. Finally, we demonstrate that cells expressing EML4-ALK V3 have increased sensitivity to microtubule poisons that interfere with mitotic spindle assembly, whereas combination treatment with paclitaxel and clinically approved ALK inhibitors leads to a synergistic response in terms of reduced survival of H2228 cells. IMPLICATIONS: This study suggests that combining the microtubule poison, paclitaxel, with targeted ALK inhibitors may provide an effective new treatment option for patients with NSCLC with tumors that express the EML4-ALK V3 oncogenic fusion. American Association for Cancer Research 2022-06-03 2022-02-25 /pmc/articles/PMC9381094/ /pubmed/35656694 http://dx.doi.org/10.1158/1541-7786.MCR-21-1010 Text en ©2022 The Authors; Published by the American Association for Cancer Research https://creativecommons.org/licenses/by/4.0/This open access article is distributed under the Creative Commons Attribution 4.0 International (CC BY 4.0) license. |
spellingShingle | Cancer Genes and Networks Lucken, Kellie O'Regan, Laura Choi, Jene Sampson, Josephina Pashley, Sarah L. Bayliss, Richard Khan, Sam Fry, Andrew M. EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity |
title | EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity |
title_full | EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity |
title_fullStr | EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity |
title_full_unstemmed | EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity |
title_short | EML4-ALK Variant 3 Promotes Mitotic Errors and Spindle Assembly Checkpoint Deficiency Leading to Increased Microtubule Poison Sensitivity |
title_sort | eml4-alk variant 3 promotes mitotic errors and spindle assembly checkpoint deficiency leading to increased microtubule poison sensitivity |
topic | Cancer Genes and Networks |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9381094/ https://www.ncbi.nlm.nih.gov/pubmed/35656694 http://dx.doi.org/10.1158/1541-7786.MCR-21-1010 |
work_keys_str_mv | AT luckenkellie eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT oreganlaura eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT choijene eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT sampsonjosephina eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT pashleysarahl eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT baylissrichard eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT khansam eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity AT fryandrewm eml4alkvariant3promotesmitoticerrorsandspindleassemblycheckpointdeficiencyleadingtoincreasedmicrotubulepoisonsensitivity |