ATAC-Seq Reveals an <i>Isl1</i> Enhancer That Regulates Sinoatrial Node Development and Function

  • Giselle Galang
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Ravi Mandla
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Hongmei Ruan
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Catherine Jung
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Tanvi Sinha
    Cardiovascular Research Institute (T.S., R.S.W., B.L.B., V.V.), University of California, San Francisco.
  • Nicole R. Stone
    Gladstone Institute of Cardiovascular Disease, San Francisco, CA (N.R.S.).
  • Roland S. Wu
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Brandon J. Mannion
    Environmental and Systems Biology Division, Lawrence Berkeley National Laboratory, CA (B.J.M., L.A.P.).
  • Prasanna K.R. Allu
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Kevin Chang
    School of Medicine (K.C.), University of California, San Francisco.
  • Ashwin Rammohan
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Marie B. Shi
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.
  • Len A. Pennacchio
    Environmental and Systems Biology Division, Lawrence Berkeley National Laboratory, CA (B.J.M., L.A.P.).
  • Brian L. Black
    Cardiovascular Research Institute (T.S., R.S.W., B.L.B., V.V.), University of California, San Francisco.
  • Vasanth Vedantham
    Cardiology Division (G.G., R.M., H.R., C.J., R.S.W., P.K.R.A., A.R., M.B.S., V.V.), University of California, San Francisco.

抄録

<jats:sec> <jats:title>Rationale:</jats:title> <jats:p> Cardiac pacemaker cells (PCs) in the sinoatrial node (SAN) have a distinct gene expression program that allows them to fire automatically and initiate the heartbeat. Although critical SAN transcription factors, including Isl1 (Islet-1), Tbx3 (T-box transcription factor 3), and Shox2 (short-stature homeobox protein 2), have been identified, the <jats:italic>cis</jats:italic> -regulatory architecture that governs PC-specific gene expression is not understood, and discrete enhancers required for gene regulation in the SAN have not been identified. </jats:p> </jats:sec> <jats:sec> <jats:title>Objective:</jats:title> <jats:p>To define the epigenetic profile of PCs using comparative ATAC-seq (assay for transposase-accessible chromatin with sequencing) and to identify novel enhancers involved in SAN gene regulation, development, and function.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods and Results:</jats:title> <jats:p> We used ATAC-seq on sorted neonatal mouse SAN to compare regions of accessible chromatin in PCs and right atrial cardiomyocytes. PC-enriched assay for transposase-accessible chromatin peaks, representing candidate SAN regulatory elements, were located near established SAN genes and were enriched for distinct sets of TF (transcription factor) binding sites. Among several novel SAN enhancers that were experimentally validated using transgenic mice, we identified a 2.9-kb regulatory element at the <jats:italic>Isl1</jats:italic> locus that was active specifically in the cardiac inflow at embryonic day 8.5 and throughout later SAN development and maturation. Deletion of this enhancer from the genome of mice resulted in SAN hypoplasia and sinus arrhythmias. The mouse SAN enhancer also directed reporter activity to the inflow tract in developing zebrafish hearts, demonstrating deep conservation of its upstream regulatory network. Finally, single nucleotide polymorphisms in the human genome that occur near the region syntenic to the mouse enhancer exhibit significant associations with resting heart rate in human populations. </jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions:</jats:title> <jats:p> (1) PCs have distinct regions of accessible chromatin that correlate with their gene expression profile and contain novel SAN enhancers, (2) <jats:italic>cis</jats:italic> -regulation of <jats:italic>Isl1</jats:italic> specifically in the SAN depends upon a conserved SAN enhancer that regulates PC development and SAN function, and (3) a corresponding human <jats:italic>ISL1</jats:italic> enhancer may regulate human SAN function. </jats:p> </jats:sec>

収録刊行物

  • Circulation Research

    Circulation Research 127 (12), 1502-1518, 2020-12-04

    Ovid Technologies (Wolters Kluwer Health)

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