An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data

ABSTRACT With the development of sequencing technology and analytic tools, studying within-species variations enhances the understanding of microbial biological processes. Nevertheless, most existing methods designed for strain-level analysis lack the capability to concurrently assess both strain pr...

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Main Authors: Boyan Zhou, Chan Wang, Gregory Putzel, Jiyuan Hu, Menghan Liu, Fen Wu, Yu Chen, Alejandro Pironti, Huilin Li
Format: Article
Language:English
Published: American Society for Microbiology 2024-11-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.01431-24
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author Boyan Zhou
Chan Wang
Gregory Putzel
Jiyuan Hu
Menghan Liu
Fen Wu
Yu Chen
Alejandro Pironti
Huilin Li
author_facet Boyan Zhou
Chan Wang
Gregory Putzel
Jiyuan Hu
Menghan Liu
Fen Wu
Yu Chen
Alejandro Pironti
Huilin Li
author_sort Boyan Zhou
collection DOAJ
description ABSTRACT With the development of sequencing technology and analytic tools, studying within-species variations enhances the understanding of microbial biological processes. Nevertheless, most existing methods designed for strain-level analysis lack the capability to concurrently assess both strain proportions and genome-wide single nucleotide variants (SNVs) across longitudinal metagenomic samples. In this study, we introduce LongStrain, an integrated pipeline for the analysis of large-scale metagenomic data from individuals with longitudinal or repeated samples. In LongStrain, we first utilize two efficient tools, Kraken2 and Bowtie2, for the taxonomic classification and alignment of sequencing reads, respectively. Subsequently, we propose to jointly model strain proportions and shared haplotypes across samples within individuals. This approach specifically targets tracking a primary strain and a secondary strain for each subject, providing their respective proportions and SNVs as output. With extensive simulation studies of a microbial community and single species, our results demonstrate that LongStrain is superior to two genotyping methods and two deconvolution methods across a majority of scenarios. Furthermore, we illustrate the potential applications of LongStrain in the real data analysis of The Environmental Determinants of Diabetes in the Young study and a gastric intestinal metaplasia microbiome study. In summary, the proposed analytic pipeline demonstrates marked statistical efficiency over the same type of methods and has great potential in understanding the genomic variants and dynamic changes at strain level. LongStrain and its tutorial are freely available online at https://github.com/BoyanZhou/LongStrain.IMPORTANCEThe advancement in DNA-sequencing technology has enabled the high-resolution identification of microorganisms in microbial communities. Since different microbial strains within species may contain extreme phenotypic variability (e.g., nutrition metabolism, antibiotic resistance, and pathogen virulence), investigating within-species variations holds great scientific promise in understanding the underlying mechanism of microbial biological processes. To fully utilize the shared genomic variants across longitudinal metagenomics samples collected in microbiome studies, we develop an integrated analytic pipeline (LongStrain) for longitudinal metagenomics data. It concurrently leverages the information on proportions of mapped reads for individual strains and genome-wide SNVs to enhance the efficiency and accuracy of strain identification. Our method helps to understand strains’ dynamic changes and their association with genome-wide variants. Given the fast-growing longitudinal studies of microbial communities, LongStrain which streamlines analyses of large-scale raw sequencing data should be of great value in microbiome research communities.
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spelling doaj-art-291455979dc0431b8efb2e35e9fc67092025-08-20T02:26:20ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972024-11-01121110.1128/spectrum.01431-24An integrated strain-level analytic pipeline utilizing longitudinal metagenomic dataBoyan Zhou0Chan Wang1Gregory Putzel2Jiyuan Hu3Menghan Liu4Fen Wu5Yu Chen6Alejandro Pironti7Huilin Li8Division of Biostatistics, Department of Population Health, New York University School of Medicine, New York, New York, USADivision of Biostatistics, Department of Population Health, New York University School of Medicine, New York, New York, USADepartment of Microbiology, New York University School of Medicine, New York, New York, USADivision of Biostatistics, Department of Population Health, New York University School of Medicine, New York, New York, USADepartment of Biological Sciences, Columbia University in the City of New York, New York, New York, USADivision of Epidemiology, Department of Population Health, New York University School of Medicine, New York, New York, USADivision of Epidemiology, Department of Population Health, New York University School of Medicine, New York, New York, USADepartment of Microbiology, New York University School of Medicine, New York, New York, USADivision of Biostatistics, Department of Population Health, New York University School of Medicine, New York, New York, USAABSTRACT With the development of sequencing technology and analytic tools, studying within-species variations enhances the understanding of microbial biological processes. Nevertheless, most existing methods designed for strain-level analysis lack the capability to concurrently assess both strain proportions and genome-wide single nucleotide variants (SNVs) across longitudinal metagenomic samples. In this study, we introduce LongStrain, an integrated pipeline for the analysis of large-scale metagenomic data from individuals with longitudinal or repeated samples. In LongStrain, we first utilize two efficient tools, Kraken2 and Bowtie2, for the taxonomic classification and alignment of sequencing reads, respectively. Subsequently, we propose to jointly model strain proportions and shared haplotypes across samples within individuals. This approach specifically targets tracking a primary strain and a secondary strain for each subject, providing their respective proportions and SNVs as output. With extensive simulation studies of a microbial community and single species, our results demonstrate that LongStrain is superior to two genotyping methods and two deconvolution methods across a majority of scenarios. Furthermore, we illustrate the potential applications of LongStrain in the real data analysis of The Environmental Determinants of Diabetes in the Young study and a gastric intestinal metaplasia microbiome study. In summary, the proposed analytic pipeline demonstrates marked statistical efficiency over the same type of methods and has great potential in understanding the genomic variants and dynamic changes at strain level. LongStrain and its tutorial are freely available online at https://github.com/BoyanZhou/LongStrain.IMPORTANCEThe advancement in DNA-sequencing technology has enabled the high-resolution identification of microorganisms in microbial communities. Since different microbial strains within species may contain extreme phenotypic variability (e.g., nutrition metabolism, antibiotic resistance, and pathogen virulence), investigating within-species variations holds great scientific promise in understanding the underlying mechanism of microbial biological processes. To fully utilize the shared genomic variants across longitudinal metagenomics samples collected in microbiome studies, we develop an integrated analytic pipeline (LongStrain) for longitudinal metagenomics data. It concurrently leverages the information on proportions of mapped reads for individual strains and genome-wide SNVs to enhance the efficiency and accuracy of strain identification. Our method helps to understand strains’ dynamic changes and their association with genome-wide variants. Given the fast-growing longitudinal studies of microbial communities, LongStrain which streamlines analyses of large-scale raw sequencing data should be of great value in microbiome research communities.https://journals.asm.org/doi/10.1128/spectrum.01431-24microbiomelongitudinal metagenomic datastrain-level analysisgenomic variantsstrain dynamics
spellingShingle Boyan Zhou
Chan Wang
Gregory Putzel
Jiyuan Hu
Menghan Liu
Fen Wu
Yu Chen
Alejandro Pironti
Huilin Li
An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data
Microbiology Spectrum
microbiome
longitudinal metagenomic data
strain-level analysis
genomic variants
strain dynamics
title An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data
title_full An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data
title_fullStr An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data
title_full_unstemmed An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data
title_short An integrated strain-level analytic pipeline utilizing longitudinal metagenomic data
title_sort integrated strain level analytic pipeline utilizing longitudinal metagenomic data
topic microbiome
longitudinal metagenomic data
strain-level analysis
genomic variants
strain dynamics
url https://journals.asm.org/doi/10.1128/spectrum.01431-24
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