Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota

Background: Gut dysbiosis has been associated with colorectal cancer (CRC), the third most prevalent cancer in the world. This study compares microbiota taxonomic and abundance results obtained by 16S rRNA gene sequencing (16S) and whole shotgun metagenomic sequencing to investigate their reliabilit...

Descripción completa

Detalles Bibliográficos
Autores: Bars Cortina, David, Ramon, Elies, Rius Sansalvador, Blanca, Guinó, Elisabet, Garcia Serrano, Ainhoa, Mach, Núria, Khannous Ileiffe, Olfat, Saus, Ester, Gabaldón, Toni, Ibáñez Sanz, Gemma, Rodríguez Alonso, Lorena, Mata, Alfredo, García Rodríguez, Ana, Obón Santacana, Mireia, Moreno Aguado, Víctor
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/215281
Acceso en línea:https://hdl.handle.net/2445/215281
Access Level:acceso abierto
Palabra clave:Microbiota intestinal
Càncer colorectal
Gastrointestinal microbiome
Colorectal cancer
id ES_56a0bb542ffd314ef128d3cd95873f21
oai_identifier_str oai:diposit.ub.edu:2445/215281
network_acronym_str ES
network_name_str España
repository_id_str
spelling Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiotaBars Cortina, DavidRamon, EliesRius Sansalvador, BlancaGuinó, ElisabetGarcia Serrano, AinhoaMach, NúriaKhannous Ileiffe, OlfatSaus, EsterGabaldón, ToniIbáñez Sanz, GemmaRodríguez Alonso, LorenaMata, AlfredoGarcía Rodríguez, AnaObón Santacana, MireiaMoreno Aguado, VíctorMicrobiota intestinalCàncer colorectalGastrointestinal microbiomeColorectal cancerBackground: Gut dysbiosis has been associated with colorectal cancer (CRC), the third most prevalent cancer in the world. This study compares microbiota taxonomic and abundance results obtained by 16S rRNA gene sequencing (16S) and whole shotgun metagenomic sequencing to investigate their reliability for bacteria profiling. The experimental design included 156 human stool samples from healthy controls, advanced (high-risk) colorectal lesion patients (HRL), and CRC cases, with each sample sequenced using both 16S and shotgun methods. We thoroughly compared both sequencing technologies at the species, genus, and family annotation levels, the abundance differences in these taxa, sparsity, alpha and beta diversities, ability to train prediction models, and the similarity of the microbial signature derived from these models. Results: As expected, the results showed that 16S detects only part of the gut microbiota community revealed by shotgun, although some genera were only profiled by 16S. The 16S abundance data was sparser and exhibited lower alpha diversity. In lower taxonomic ranks, shotgun and 16S highly differed, partially due to a disagreement in reference databases. When considering only shared taxa, the abundance was positively correlated between the two strategies. We also found a moderate correlation between the shotgun and 16S alpha-diversity measures, as well as their PCoAs. Regarding the machine learning models, only some of the shotgun models showed some degree of predictive power in an independent test set, but we could not demonstrate a clear superiority of one technology over the other. Microbial signatures from both sequencing techniques revealed taxa previously associated with CRC development, e.g., Parvimonas micra. Conclusions: Shotgun and 16S sequencing provide two different lenses to examine microbial communities. While we have demonstrated that they can unravel common patterns (including microbial signatures), shotgun often gives a more detailed snapshot than 16S, both in depth and breadth. Instead, 16S will tend to show only part of the picture, giving greater weight to dominant bacteria in a sample. Therefore, we recommend choosing one or another sequencing technique before launching a study. Specifically, shotgun sequencing is preferred for stool microbiome samples and in-depth analyses, while 16S is more suitable for tissue samples and studies with targeted aims.Springer Science and Business Media LLC2024info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/215281Articles publicats en revistes (Ciències Clíniques)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.1186/s12864-024-10621-7BMC Genomics, 2024, vol. 25, num. 1https://doi.org/10.1186/s12864-024-10621-7cc by (c) Bars Cortina, David et al., 2024http://creativecommons.org/licenses/by/3.0/es/info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/2152812026-05-27T06:46:51Z
dc.title.none.fl_str_mv Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
title Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
spellingShingle Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
Bars Cortina, David
Microbiota intestinal
Càncer colorectal
Gastrointestinal microbiome
Colorectal cancer
title_short Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
title_full Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
title_fullStr Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
title_full_unstemmed Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
title_sort Comparison between 16S rRNA and shotgun sequencing in colorectal cancer, advanced colorectal lesions, and healthy human gut microbiota
dc.creator.none.fl_str_mv Bars Cortina, David
Ramon, Elies
Rius Sansalvador, Blanca
Guinó, Elisabet
Garcia Serrano, Ainhoa
Mach, Núria
Khannous Ileiffe, Olfat
Saus, Ester
Gabaldón, Toni
Ibáñez Sanz, Gemma
Rodríguez Alonso, Lorena
Mata, Alfredo
García Rodríguez, Ana
Obón Santacana, Mireia
Moreno Aguado, Víctor
author Bars Cortina, David
author_facet Bars Cortina, David
Ramon, Elies
Rius Sansalvador, Blanca
Guinó, Elisabet
Garcia Serrano, Ainhoa
Mach, Núria
Khannous Ileiffe, Olfat
Saus, Ester
Gabaldón, Toni
Ibáñez Sanz, Gemma
Rodríguez Alonso, Lorena
Mata, Alfredo
García Rodríguez, Ana
Obón Santacana, Mireia
Moreno Aguado, Víctor
author_role author
author2 Ramon, Elies
Rius Sansalvador, Blanca
Guinó, Elisabet
Garcia Serrano, Ainhoa
Mach, Núria
Khannous Ileiffe, Olfat
Saus, Ester
Gabaldón, Toni
Ibáñez Sanz, Gemma
Rodríguez Alonso, Lorena
Mata, Alfredo
García Rodríguez, Ana
Obón Santacana, Mireia
Moreno Aguado, Víctor
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Microbiota intestinal
Càncer colorectal
Gastrointestinal microbiome
Colorectal cancer
topic Microbiota intestinal
Càncer colorectal
Gastrointestinal microbiome
Colorectal cancer
description Background: Gut dysbiosis has been associated with colorectal cancer (CRC), the third most prevalent cancer in the world. This study compares microbiota taxonomic and abundance results obtained by 16S rRNA gene sequencing (16S) and whole shotgun metagenomic sequencing to investigate their reliability for bacteria profiling. The experimental design included 156 human stool samples from healthy controls, advanced (high-risk) colorectal lesion patients (HRL), and CRC cases, with each sample sequenced using both 16S and shotgun methods. We thoroughly compared both sequencing technologies at the species, genus, and family annotation levels, the abundance differences in these taxa, sparsity, alpha and beta diversities, ability to train prediction models, and the similarity of the microbial signature derived from these models. Results: As expected, the results showed that 16S detects only part of the gut microbiota community revealed by shotgun, although some genera were only profiled by 16S. The 16S abundance data was sparser and exhibited lower alpha diversity. In lower taxonomic ranks, shotgun and 16S highly differed, partially due to a disagreement in reference databases. When considering only shared taxa, the abundance was positively correlated between the two strategies. We also found a moderate correlation between the shotgun and 16S alpha-diversity measures, as well as their PCoAs. Regarding the machine learning models, only some of the shotgun models showed some degree of predictive power in an independent test set, but we could not demonstrate a clear superiority of one technology over the other. Microbial signatures from both sequencing techniques revealed taxa previously associated with CRC development, e.g., Parvimonas micra. Conclusions: Shotgun and 16S sequencing provide two different lenses to examine microbial communities. While we have demonstrated that they can unravel common patterns (including microbial signatures), shotgun often gives a more detailed snapshot than 16S, both in depth and breadth. Instead, 16S will tend to show only part of the picture, giving greater weight to dominant bacteria in a sample. Therefore, we recommend choosing one or another sequencing technique before launching a study. Specifically, shotgun sequencing is preferred for stool microbiome samples and in-depth analyses, while 16S is more suitable for tissue samples and studies with targeted aims.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/215281
url https://hdl.handle.net/2445/215281
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1186/s12864-024-10621-7
BMC Genomics, 2024, vol. 25, num. 1
https://doi.org/10.1186/s12864-024-10621-7
dc.rights.none.fl_str_mv cc by (c) Bars Cortina, David et al., 2024
http://creativecommons.org/licenses/by/3.0/es/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc by (c) Bars Cortina, David et al., 2024
http://creativecommons.org/licenses/by/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer Science and Business Media LLC
publisher.none.fl_str_mv Springer Science and Business Media LLC
dc.source.none.fl_str_mv Articles publicats en revistes (Ciències Clíniques)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869408393368698880
score 15,811543