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Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells
Mismatch repair (MMR)-deficient or microsatellite instability (MSI) colorectal cancer includes two subtypes; Lynch syndrome and sporadic MSI cancer, both of which generate multiple neoantigens due to unrepaired mutations. Although such patients respond very well to immune checkpoint therapy, their d...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6331025/ https://www.ncbi.nlm.nih.gov/pubmed/30680068 http://dx.doi.org/10.18632/oncotarget.26495 |
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author | Yamaura, Tadayoshi Miyoshi, Hiroyuki Maekawa, Hisatsugu Morimoto, Tomonori Yamamoto, Takehito Kakizaki, Fumihiko Higasa, Koichiro Kawada, Kenji Matsuda, Fumihiko Sakai, Yoshiharu Taketo, M. Mark |
author_facet | Yamaura, Tadayoshi Miyoshi, Hiroyuki Maekawa, Hisatsugu Morimoto, Tomonori Yamamoto, Takehito Kakizaki, Fumihiko Higasa, Koichiro Kawada, Kenji Matsuda, Fumihiko Sakai, Yoshiharu Taketo, M. Mark |
author_sort | Yamaura, Tadayoshi |
collection | PubMed |
description | Mismatch repair (MMR)-deficient or microsatellite instability (MSI) colorectal cancer includes two subtypes; Lynch syndrome and sporadic MSI cancer, both of which generate multiple neoantigens due to unrepaired mutations. Although such patients respond very well to immune checkpoint therapy, their diagnosis can be confused by low quality DNA samples owing to formalin fixation and/or low cancer cell content. Here we prepared high-quality DNA samples from in vitro-cultured cancer spheroids that consisted of the pure cell population. We evaluated their diagnostic power by on-chip electrophoresis, mutational burden assessment, and direct sequencing. Because formalin-fixed paraffin-embedded (FFPE) tissues are widely used as the DNA source, we compared such samples with spheroid DNA. Additionally, we performed immunohistochemistry (IHC) for MMR proteins on spheroids as well as primary tumor sections. Of 111 cases of colorectal cancer patients, we found seven MSI-high cases in which all diagnostic results agreed on spheroid-based assays, whereas the results with the FFPE DNA were less reliable though analyzable. Importantly, there was an MSS case that appeared as MSI by IHC on primary tumor sections. Based on these results, we propose to employ cultured cancer spheroids as the source of both DNA and IHC specimens for more reliable clinical diagnosis. |
format | Online Article Text |
id | pubmed-6331025 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-63310252019-01-24 Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells Yamaura, Tadayoshi Miyoshi, Hiroyuki Maekawa, Hisatsugu Morimoto, Tomonori Yamamoto, Takehito Kakizaki, Fumihiko Higasa, Koichiro Kawada, Kenji Matsuda, Fumihiko Sakai, Yoshiharu Taketo, M. Mark Oncotarget Research Paper Mismatch repair (MMR)-deficient or microsatellite instability (MSI) colorectal cancer includes two subtypes; Lynch syndrome and sporadic MSI cancer, both of which generate multiple neoantigens due to unrepaired mutations. Although such patients respond very well to immune checkpoint therapy, their diagnosis can be confused by low quality DNA samples owing to formalin fixation and/or low cancer cell content. Here we prepared high-quality DNA samples from in vitro-cultured cancer spheroids that consisted of the pure cell population. We evaluated their diagnostic power by on-chip electrophoresis, mutational burden assessment, and direct sequencing. Because formalin-fixed paraffin-embedded (FFPE) tissues are widely used as the DNA source, we compared such samples with spheroid DNA. Additionally, we performed immunohistochemistry (IHC) for MMR proteins on spheroids as well as primary tumor sections. Of 111 cases of colorectal cancer patients, we found seven MSI-high cases in which all diagnostic results agreed on spheroid-based assays, whereas the results with the FFPE DNA were less reliable though analyzable. Importantly, there was an MSS case that appeared as MSI by IHC on primary tumor sections. Based on these results, we propose to employ cultured cancer spheroids as the source of both DNA and IHC specimens for more reliable clinical diagnosis. Impact Journals LLC 2018-12-25 /pmc/articles/PMC6331025/ /pubmed/30680068 http://dx.doi.org/10.18632/oncotarget.26495 Text en Copyright: © 2018 Yamaura et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/) 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Paper Yamaura, Tadayoshi Miyoshi, Hiroyuki Maekawa, Hisatsugu Morimoto, Tomonori Yamamoto, Takehito Kakizaki, Fumihiko Higasa, Koichiro Kawada, Kenji Matsuda, Fumihiko Sakai, Yoshiharu Taketo, M. Mark Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells |
title | Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells |
title_full | Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells |
title_fullStr | Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells |
title_full_unstemmed | Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells |
title_short | Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells |
title_sort | accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality dna samples from cultured stem cells |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6331025/ https://www.ncbi.nlm.nih.gov/pubmed/30680068 http://dx.doi.org/10.18632/oncotarget.26495 |
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