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A ChIP-seq-Defined Genome-Wide Map of MEF2C Binding Reveals Inflammatory Pathways Associated with Its Role in Bone Density Determination

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Abstract

Genome-wide association studies (GWAS) have demonstrated that genetic variation at the MADS box transcription enhancer factor 2, polypeptide C (MEF2C) locus is robustly associated with bone mineral density, primarily at the femoral neck. MEF2C is a transcription factor known to operate via the Wnt signaling pathway. Our hypothesis was that MEF2C regulates the expression of a set of molecular pathways critical to skeletal function. Drawing on our laboratory and bioinformatic experience with ChIP-seq, we analyzed ChIP-seq data for MEF2C available via the ENCODE project to gain insight in to its global genomic binding pattern. We aligned the ChIP-seq data generated for GM12878 (an established lymphoblastoid cell line) and, using the analysis package HOMER, a total of 17,611 binding sites corresponding to 8,118 known genes were observed. We then performed a pathway analysis of the gene list using Ingenuity. At 5 kb, the gene list yielded ‘EIF2 Signaling’ as the most significant annotation, with a P value of 5.01 × 10−26. Moving further out, this category remained the top pathway at 50 and 100 kb, then dropped to just second place at 500 kb and beyond by ‘Molecular Mechanisms of Cancer’. In addition, at 50 kb and beyond ‘RANK Signaling in Osteoclasts’ was a consistent feature and resonates with the main general finding from GWAS of bone density. We also observed that MEF2C binding sites were significantly enriched primarily near inflammation associated genes identified from GWAS; indeed, a similar enrichment for inflammation genes has been reported previously using a similar approach for the vitamin D receptor, an established key regulator of bone turnover. Our analyses point to known connective tissue and skeletal processes but also provide novel insights in to networks involved in skeletal regulation. The fact that a specific GWAS category is enriched points to a possible role of inflammation through which it impacts bone mineral density.

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Acknowledgments

This work was supported by Institutional Development Funds from the Children’s Hospital of Philadelphia and a Penn Center for Musculoskeletal Disorders pilot award.

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Authors and Affiliations

  1. Division of Human Genetics, The Children’s Hospital of Philadelphia Research Center, Rm 1216, 3615 Civic Center Blvd, Philadelphia, PA, 19104, USA

    Matthew E. Johnson, Sandra Deliard, Qianghua Xia & Struan F. A. Grant

  2. School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Fengchang Zhu & Kurt D. Hankenson

  3. Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, Philadelphia, PA, 19104, USA

    Andrew D. Wells

  4. Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Struan F. A. Grant

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  1. Matthew E. Johnson
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Johnson, M.E., Deliard, S., Zhu, F. et al. A ChIP-seq-Defined Genome-Wide Map of MEF2C Binding Reveals Inflammatory Pathways Associated with Its Role in Bone Density Determination. Calcif Tissue Int 94, 396–402 (2014). https://doi.org/10.1007/s00223-013-9824-5

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