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RNA-sequencing indicates immune cell signaling and inflammatory gene expression in cardiac fibroblasts increases with developmental age

View ORCID ProfileLuke R. Perreault, Thanh T. Le, View ORCID ProfileMadeleine J. Oudin, Lauren D. Black III
doi: https://doi.org/10.1101/2021.03.01.433442
Luke R. Perreault
aDepartment of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155
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Thanh T. Le
aDepartment of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155
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Madeleine J. Oudin
aDepartment of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155
bCellular, Molecular, and Developmental Biology Program, Tufts Graduate School of Biomedical Sciences, Tufts University School of Medicine, 145 Harrison Avenue, Boston, MA 02111
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Lauren D. Black III
aDepartment of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155
bCellular, Molecular, and Developmental Biology Program, Tufts Graduate School of Biomedical Sciences, Tufts University School of Medicine, 145 Harrison Avenue, Boston, MA 02111
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  • For correspondence: Lauren.Black@tufts.edu
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Abstract

Background Cardiac fibroblasts are responsible for extracellular matrix turnover and repair in the cardiac environment and serve to help facilitate immune responses. However, it is well established that they have significant phenotypic heterogeneity with respect to location, physiological conditions, and developmental age. The goal of this study was to provide an in-depth transcriptomic profile of cardiac fibroblasts derived from rat hearts at fetal, neonatal, and adult developmental ages to ascertain variations in gene expression that may drive functional differences in these cells at these specific stages of development.

Results We performed RNA-seq of cardiac fibroblasts isolated from fetal, neonatal, and adult rats was performed and compared to the rat genome. Principal component analysis of RNA-seq data suggested data variance was predominantly due to developmental age. Differential expression and Gene set enrichment analysis against Gene Ontology and Kyoto Encyclopedia of Genes and Genomes datasets indicated an array of differences across developmental ages, including significant decreases in cardiac development and cardiac function-associated genes with age, and a significant increase in immune and inflammatory-associated functions - particularly immune cell signaling, and cytokine and chemokine production - with respect to increasing developmental age.

Conclusion These results reinforce established evidence of diverse phenotypic heterogeneity of fibroblasts with respect to developmental age. Further, based on our analysis of gene expression, age-specific alterations in cardiac fibroblasts may play a crucial role in observed differences in cardiac inflammation and immune response observed across developmental ages.

Competing Interest Statement

The authors have declared no competing interest.

  • Abbreviations

    cDNA
    complementary deoxyribonucleic acid
    CF
    cardiac fibroblast
    CM
    cardiomyocyte
    DAPI
    4’,6-diamidino-2-phenylindole
    DDR2
    discoidin domain receptor 2
    DNA
    deoxyribonucleic acid
    ECM
    extracellular matrix
    FBS
    fetal bovine serum
    GAPDH
    glyceraldehyde 3-phosphate dehydrogenase
    GO
    gene ontology
    IL
    interleukin
    KEGG
    Kyoto Encyclopedia of Genes and Genomes
    logFC
    log fold change
    LPS
    lipopolysaccharide
    MMP
    matrix metalloproteinase
    MYH
    myosin heavy chain
    mRNA
    messenger ribonucleic acid
    PBS
    phosphate buffered saline
    PCA
    principal component analysis
    RNA
    ribonucleic acid
    RNA-seq
    RNA sequencing
    RT-qPCR
    reverse transcriptase quantitative polymerase chain reaction
  • Copyright 
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    Posted March 01, 2021.
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    RNA-sequencing indicates immune cell signaling and inflammatory gene expression in cardiac fibroblasts increases with developmental age
    Luke R. Perreault, Thanh T. Le, Madeleine J. Oudin, Lauren D. Black III
    bioRxiv 2021.03.01.433442; doi: https://doi.org/10.1101/2021.03.01.433442
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    RNA-sequencing indicates immune cell signaling and inflammatory gene expression in cardiac fibroblasts increases with developmental age
    Luke R. Perreault, Thanh T. Le, Madeleine J. Oudin, Lauren D. Black III
    bioRxiv 2021.03.01.433442; doi: https://doi.org/10.1101/2021.03.01.433442

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