Cardiac Lineage-Specific Molecular Mechanisms of Heart Failure

项目摘要 心脏由多种不同的心脏细胞类型组成，这些细胞类型形成心脏结构， 维持心脏功能。这些细胞群不仅包括心肌细胞、心脏成纤维细胞， 心外膜细胞、内皮/内皮细胞和平滑肌细胞，以及更特化的细胞类型 包括心脏瓣膜，心脏传导系统等。因此，这些细胞的调节维持 类型对最佳心脏性能至关重要，破坏特定细胞谱系的功能可导致 不同的心脏病，包括心力衰竭（HF），这是发病率和死亡率的主要主要原因 国际吧然而，在HF过程中，哪些特定的细胞谱系受到影响，基因调控是如何进行的？ 控制遗传程序并指导其病理结果的神经网络是关键的生物医学问题 我们在母公司R 01 HL 156576赠款中寻求解决。为此，这项拨款建议主要包括： 集中于分析心腔（即，心室和心房），但不包括心脏瓣膜，心脏瓣膜可以 在HF中有缺陷和患病，并且在某些情况下可能是HF的原因。因此，为了扩大我们的努力， 检查参与体内衰竭和非衰竭人类心脏（包括心脏瓣膜）的所有细胞类型， 我们提出实施联合的单细胞/核（sc/sn）RNA-seq和ATAC-seq技术（即，单细胞 多组学）不仅在我们的母基金中最初提出的心腔上， 瓣膜，尤其是二尖瓣，以回应特别关注通知（NOSI）：行政 鼓励心脏瓣膜病研究的补充文件（CAROL法案，NOT-HL-23-078）。除了 识别不同的CV细胞类型及其相关的转录谱和染色质景观，我们进一步 试图阐明细胞类型特异性顺式调节元件（克雷斯）之间的相互作用， GRN控制克雷斯如何指导这些细胞类型在衰竭的心脏（二尖瓣）瓣膜中的基因表达。 和不会衰竭的心脏此外，由于心脏瓣膜的结构形式对于调节其功能至关重要， 功能，我们建议进一步研究心脏瓣膜中已识别细胞类型的空间组织， 特别是二尖瓣，具有和不具有心力衰竭和心脏瓣膜缺陷/疾病的人心脏。 因此，这项补充我们原来的母基金的总体目标是产生单细胞多组学， 心脏瓣膜，特别是二尖瓣的空间成像数据，来自一个大的，表型良好的队列， 患有和不患有心力衰竭和二尖瓣缺陷/疾病的患者。然后我们将创建全面的 细胞类型特异性基因调控的图谱，包括整合的基因组图谱和所有 细胞和每种细胞类型中的顺式调节程序。

Project Summary The heart consists of a multitude of diverse cardiac cell types that form cardiac structures critical for maintaining heart function. These cell populations include not only cardiomyocytes, cardiac fibroblasts, epicardial cells, endothelial/endocardial cells and smooth muscle cells, but also more specialized cell types comprising the cardiac valves, cardiac conduction system, etc. Thus, regulated maintenance of these cell types is crucial for optimal heart performance, and disrupting the function of specific cell lineages can result in distinct heart diseases including heart failure (HF), which is a major leading cause of morbidity and mortality worldwide. However, what are the specific cell lineages affected during HF and how do gene regulatory networks (GRNs) control genetic programs that direct their pathologic outcomes are key biomedical questions that we seek to address in our parent R01HL156576 grant. Toward this end, this grant proposal has mainly focused on analyzing the cardiac chambers (i.e., ventricles and atria) but not cardiac valves, which can be defective and diseased in HF and in some cases can be the cause of HF. Thus, to expand our efforts and examine all cell-types participating in failing and non-failing human hearts in vivo including the cardiac valves, we propose to implement joint single cell/nuclear (sc/sn) RNA-seq and ATAC-seq technologies (i.e., single cell multi-omics) on not only the cardiac chambers as originally proposed in our parent grant but also the cardiac valves, particularly mitral valve, in response to the Notice of Special Interest (NOSI): Administrative Supplements to Encourage Research in Valvular Heart Disease (CAROL Act, NOT-HL-23-078). In addition to identifying distinct CV cell-types and their related transcriptional profiles and chromatin landscape, we further seek to elucidate the interactions between cell-type specific cis-regulatory elements (CREs), which mediate the GRNs that control how CREs direct gene expression of these cell types in the cardiac (mitral) valves of failing and non-failing hearts. Furthermore, because the structural form of the cardiac valve is critical for regulating its function, we propose to further investigate the spatial organization of identified cell types in the cardiac valves, particularly mitral valves, of human hearts with and without heart failure and cardiac valve defects/disease. Thus, the overall goal of this supplement to our original parent grant is to generate single-cell multi-omics and spatial imaging data of cardiac valves, particularly mitral valves, from a large, well-phenotyped cohort of patients with and without heart failure and mitral valve defects/disease. We will then create comprehensive maps of cell-type specific gene regulation including integrated genomic profiles and spatial localization of all cells and cis-regulatory programs in each cell type.