The HBCLL center, in the CvDC, collaborates with the PCGC to form the twin pillars of the B2B Program. The B2B seeks to define the complex molecular and cell interactions that are required for normal heart development, to define genetic perturbations that cause congenital heart disease (CHD), and to elucidate the mechanisms by which these CHD mutations disrupt heart development. We will address the following unanswered questions: What are the DNA elements with which these global transcriptional regulators interact? Might mutations in these DNA elements also cause CHD? What are the genes with altered expression and how does disruption of physiologic temporal-spatial expression result in CHD? The answers to these questions will significantly expand our understanding of cardiac development as well as fundamental processes involved in cell lineage commitment, differentiation and ultimately organogenesis. Our overarching goal is to establish a blueprint of the transcriptional regulatory elements that direct cardiogenesis, and to use this information to identify the rare variants that cause CHD and to elucidate how these disrupt heart development. The HBCLL will use next generation sequencing technologies to map both chromatin structure and gene expression during cardiac development. The new maps will consist of spatiotemporal and cell type specific assessment of transcript abundance (Aim 3) and structure (Aim 1,2). We also propose a series of collaborative experiments, with the PCGC and other CvDC centers, to distinguish pathogenic human sequence variants from benign variants. We suggest that each group in CvDC and PCGC centers focus on variants that are appropriate to their expertise. As CHD-associated variants will likely impair multiple distinct developmental pathways this `bundling' of variants will facilitate analyses. For example, one group will assess the pathogenicity of approximately 150 recently discovered variants that are in transcriptional regulatory and chromatin modifying genes. Similarly, another set of variants may be involved in altering RNA splicing. Another set of variants will likely fall in non-coding regulatory regions an land on our regulatory element map; these could be characterized using cell or mouse-based models (Aim 3). Characterization of these human variants in human iPS cells and animal models will provide novel insights into developmental networks and regulatory blueprints and their perturbations in CHD. In summary we propose experiments directed towards understanding cardiovascular development and its perturbation in CHD. We have 4 specific aims: 1) Define the RNA transcriptome in human normal and CHD tissues; 2) Complete the murine cardiac transcriptome map and assess mechanisms of cardiac chamber formation by single cell analyses; 3) Annotate regulatory elements and their interacting transcriptional regulatory proteins in human and mouse genomes and 4) Perform functional analyses of human genetic variants and critical transcriptional regulatory nodes (Collaborative Project).
Congenital heart disease, a significant health concern, is likely caused by defects in heart development. We plan to map all of the steps by which mouse and human hearts are formed. In collaboration with other researchers and physicians we will use this developmental map to identify inherited defects that cause malformed hearts.
| Bressan, Michael; Henley, Trevor; Louie, Jonathan D et al. (2018) Dynamic Cellular Integration Drives Functional Assembly of the Heart's Pacemaker Complex. Cell Rep 23:2283-2291 |
| Guo, Yuxuan; Pu, William T (2018) Genetic Mosaics for Greater Precision in Cardiovascular Research. Circ Res 123:27-29 |
| Osterwalder, Marco; Barozzi, Iros; Tissières, Virginie et al. (2018) Enhancer redundancy provides phenotypic robustness in mammalian development. Nature 554:239-243 |
| Dickel, Diane E; Ypsilanti, Athena R; Pla, Ramón et al. (2018) Ultraconserved Enhancers Are Required for Normal Development. Cell 172:491-499.e15 |
| Anderson, Courtney M; Hu, Jianxin; Thomas, Reuben et al. (2017) Cooperative activation of cardiac transcription through myocardin bridging of paired MEF2 sites. Development 144:1235-1241 |
| Galdos, Francisco X; Guo, Yuxuan; Paige, Sharon L et al. (2017) Cardiac Regeneration: Lessons From Development. Circ Res 120:941-959 |
| Gompers, Andrea L; Su-Feher, Linda; Ellegood, Jacob et al. (2017) Germline Chd8 haploinsufficiency alters brain development in mouse. Nat Neurosci 20:1062-1073 |
| Zhou, Pingzhu; Gu, Fei; Zhang, Lina et al. (2017) Mapping cell type-specific transcriptional enhancers using high affinity, lineage-specific Ep300 bioChIP-seq. Elife 6: |
| Saddic, Louis A; Sigurdsson, Martin I; Chang, Tzuu-Wang et al. (2017) The Long Noncoding RNA Landscape of the Ischemic Human Left Ventricle. Circ Cardiovasc Genet 10: |
| Ai, Shanshan; Yu, Xianhong; Li, Yumei et al. (2017) Divergent Requirements for EZH1 in Heart Development Versus Regeneration. Circ Res 121:106-112 |
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