Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle

RNA-sequencing (RNA-seq) is an important tool to explore molecular mechanisms of disease. Technological advances mean this can be performed at the single-cell level, but the large sample sizes needed in clinical studies are currently prohibitively expensive and complex. Deconvolution of bulk RNA-seq...

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Main Authors: Marcella Conning-Rowland, Chew W. Cheng, Oliver Brown, Marilena Giannoudi, Eylem Levelt, Lee D. Roberts, Kathryn J. Griffin, Richard M. Cubbon
Format: Article
Language:English
Published: Elsevier 2025-02-01
Series:Heliyon
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Online Access:http://www.sciencedirect.com/science/article/pii/S2405844025008795
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author Marcella Conning-Rowland
Chew W. Cheng
Oliver Brown
Marilena Giannoudi
Eylem Levelt
Lee D. Roberts
Kathryn J. Griffin
Richard M. Cubbon
author_facet Marcella Conning-Rowland
Chew W. Cheng
Oliver Brown
Marilena Giannoudi
Eylem Levelt
Lee D. Roberts
Kathryn J. Griffin
Richard M. Cubbon
author_sort Marcella Conning-Rowland
collection DOAJ
description RNA-sequencing (RNA-seq) is an important tool to explore molecular mechanisms of disease. Technological advances mean this can be performed at the single-cell level, but the large sample sizes needed in clinical studies are currently prohibitively expensive and complex. Deconvolution of bulk RNA-seq offers an opportunity to bridge this gap by defining the cell lineage composition of samples. This approach is widely used in immunology studies, but currently there are no validated pipelines for researchers analysing human myocardium or skeletal muscle. Here, we describe the application and in silico validation of two pipelines to deconvolute human right atrium, left ventricle and skeletal muscle bulk RNA-seq data. Specifically, we have defined the major cell lineages of these tissues using single cell/nucleus RNA-seq data from the Heart Cell Atlas, which are then applied during deconvolution using the CIBERSORTx or BayesPrism deconvolution packages. Both pipelines gave robust estimates of the proportion of all major cell lineages in these tissues. We demonstrate their value in defining age- and sex-differences in tissue composition using bulk RNA-seq data from the GTEx consortium. Our validated pipelines can be rapidly applied by researchers working with existing or novel bulk RNA-seq of myocardium or skeletal muscle to gain novel insights.
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spelling doaj-art-23efa1d875d74abb8267dbe0c3d9aa8b2025-08-20T02:13:20ZengElsevierHeliyon2405-84402025-02-01114e4249910.1016/j.heliyon.2025.e42499Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscleMarcella Conning-Rowland0Chew W. Cheng1Oliver Brown2Marilena Giannoudi3Eylem Levelt4Lee D. Roberts5Kathryn J. Griffin6Richard M. Cubbon7Leeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomLeeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomLeeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomLeeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomLeeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomLeeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomLeeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomCorresponding author. LIGHT laboratories 7.04, Clarendon Way, Leeds, LS2 9JT, United Kingdom.; Leeds Institute of Cardiovascular and Metabolic Medicine, The University of Leeds, Leeds, United KingdomRNA-sequencing (RNA-seq) is an important tool to explore molecular mechanisms of disease. Technological advances mean this can be performed at the single-cell level, but the large sample sizes needed in clinical studies are currently prohibitively expensive and complex. Deconvolution of bulk RNA-seq offers an opportunity to bridge this gap by defining the cell lineage composition of samples. This approach is widely used in immunology studies, but currently there are no validated pipelines for researchers analysing human myocardium or skeletal muscle. Here, we describe the application and in silico validation of two pipelines to deconvolute human right atrium, left ventricle and skeletal muscle bulk RNA-seq data. Specifically, we have defined the major cell lineages of these tissues using single cell/nucleus RNA-seq data from the Heart Cell Atlas, which are then applied during deconvolution using the CIBERSORTx or BayesPrism deconvolution packages. Both pipelines gave robust estimates of the proportion of all major cell lineages in these tissues. We demonstrate their value in defining age- and sex-differences in tissue composition using bulk RNA-seq data from the GTEx consortium. Our validated pipelines can be rapidly applied by researchers working with existing or novel bulk RNA-seq of myocardium or skeletal muscle to gain novel insights.http://www.sciencedirect.com/science/article/pii/S2405844025008795RNA-SeqDeconvolutionMyocardiumSkeletal muscleAgeSex
spellingShingle Marcella Conning-Rowland
Chew W. Cheng
Oliver Brown
Marilena Giannoudi
Eylem Levelt
Lee D. Roberts
Kathryn J. Griffin
Richard M. Cubbon
Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle
Heliyon
RNA-Seq
Deconvolution
Myocardium
Skeletal muscle
Age
Sex
title Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle
title_full Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle
title_fullStr Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle
title_full_unstemmed Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle
title_short Application of CIBERSORTx and BayesPrism to deconvolution of bulk RNA-seq data from human myocardium and skeletal muscle
title_sort application of cibersortx and bayesprism to deconvolution of bulk rna seq data from human myocardium and skeletal muscle
topic RNA-Seq
Deconvolution
Myocardium
Skeletal muscle
Age
Sex
url http://www.sciencedirect.com/science/article/pii/S2405844025008795
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