Elucidating the Role of Multinuclearity in Healthy and Diseased Mammalian Cardiomyocytes

阐明多核在健康和患病哺乳动物心肌细胞中的作用

基本信息

批准号：
10555524
负责人：
Christoph Daniel Rau
金额：
$ 48.57万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10555524
关键词：
Adult Affect Age Algorithms Apoptosis Bioinformatics Blood Cardiac Cardiac Myocytes Cardiac Volume Cardiac health Cell Nucleus Cell division Cells Code Collaborations Computational algorithm DNA Data Development Diagnostics Research Disease Elements Gene Expression Genes Genetic Genetic Transcription Genetic Variation Genomics Goals Grant Heart Heart Injuries Heart failure Hypertrophy Incidence Infarction Laboratory Research Link Literature Mononuclear Mus Muscle Cells Myocardial Infarction Nature Nuclear Phenotype Polyploidy Population Population Heterogeneity Predisposition Proliferating Publishing Regenerative capacity Research Research Personnel Resistance Role Severities Site Spatial Distribution Stress Technology Therapeutic Transcript Variant Visual Work cell type combat comparative effective therapy heart cell heart function image reconstruction mortality new technology next generation sequencing novel novel strategies novel therapeutics overexpression response single nucleus RNA-sequencing single-cell RNA sequencing skills stressor three dimensional structure transcriptome transcriptome sequencing transcriptomics trend

Adult Affect Age Algorithms Apoptosis Bioinformatics Blood Cardiac Cardiac Myocytes Cardiac Volume Cardiac health Cell Nucleus Cell division Cells Code Collaborations Computational algorithm DNA Data Development Diagnostics Research Disease Elements Gene Expression Genes Genetic Genetic Transcription Genetic Variation Genomics Goals Grant Heart Heart Injuries Heart failure Hypertrophy Incidence Infarction Laboratory Research Link Literature Mononuclear Mus Muscle Cells Myocardial Infarction Nature Nuclear Phenotype Polyploidy Population Population Heterogeneity Predisposition Proliferating Publishing Regenerative capacity Research Research Personnel Resistance Role Severities Site Spatial Distribution Stress Technology Therapeutic Transcript Variant Visual Work cell type combat comparative effective therapy heart cell heart function image reconstruction mortality new technology next generation sequencing novel novel strategies novel therapeutics overexpression response single nucleus RNA-sequencing single-cell RNA sequencing skills stressor three dimensional structure transcriptome transcriptome sequencing transcriptomics trend

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项目摘要

PROJECT SUMMARY Heart Failure has resisted the downward trend in mortality seen in other diseases and new therapies are needed. Heart failure is driven by the hypertrophy and loss of cardiomyocytes, which comprise the bulk of the heart by volume and provide the main contractile force necessary to move blood throughout the body. Over 90% of cardiomyocytes in mammalian species are polyploidal and/or multinucleated. These extra copies of DNA have been studied in the context of the low regenerative capacity of the heart, yet comparatively little effort has focused on other potential adaptive or maladaptive effects of this increased nuclearity. Historically, the large size of the mature cardiomyocyte has precluded its analysis using unbiased single cell RNA sequencing. Research has focused on RNAseq in isolated nuclei, which by its very nature lacks information about nuclearity within the studied hearts. Recently, new technology has allowed for unbiased, high-throughput single cell RNAseq of whole, mature cardiomyocytes. When the results of these single cell studies are compared to the isolated nuclei studies, researchers find that, using the same clustering algorithms, there are approximately twice as many distinct cellular transcriptome clusters as there are distinct nuclear transcriptome clusters. This result suggests the existence of a form of ‘nuclear code’ in which a multinucleated cell’s transcriptome is ‘encoded’ by nuclei drawn from two or more nuclear clusters, a result supported by observations from other multinucleated cell populations. These encoded cells could display differences in contractility, resistance to apoptosis, or even proliferative potential, with significant implications for cardiac function and the development of heart failure. The goal of this Steven I. Katz proposal is to 1) Utilize cutting edge single cell and single nucleus sequencing technology to identify and confirm this nuclear encoding, 2) Explore the spatial distribution of mono and multi- nucleated cells across the heart, and 3) Identify how this encoding responds to genetic and environmental perturbations in healthy and failing hearts. As requested by the mechanism, in this proposal we set forth a plan for an ambitious new direction for our laboratory’s research, built upon rigorous work documented in the literature and supported by collaborators and subject matter experts. The proposal combines bioinformatic, imaging, and next generation sequencing approaches to identify how changes in the nuclear code of multinucleated cardiomyocytes leads to differences in response to cardiac injury and stress. Upon completion, this grant will result in a more complete understanding of the role of multinuclearity and polyploidy in the mammalian heart, with subsequent revelation of numerous new avenues of research for diagnostic and therapeutic approaches to combat heart failure.

项目摘要心力衰竭抵制了其他疾病中死亡率的下降趋势，而新疗法是需要。心力衰竭是由肥大和心肌丧失驱动的，其中大部分心脏按体积为单位，并提供在整个身体中移动血液所需的主要收缩力。超过哺乳动物物种中90％的心肌细胞是多倍体和/或多核核细胞。这些额外的副本 DNA在心脏的再生能力较低的背景下已研究，但相对较少努力集中在这种增加的核性的其他潜在适应性或不良适应性作用上。从历史上看，大尺寸成熟的心肌细胞已使用公正的单细胞排除了其分析 RNA测序。研究集中于孤立核的RNASEQ，从本质上就缺乏有关核性心脏内核性的信息。最近，新技术允许公正，整体成熟心肌细胞的高通量单细胞RNASEQ。当这些单个单元的结果研究人员将研究与孤立的核研究进行了比较，研究人员发现，使用相同的聚类算法，大约有不同的细胞转录组簇的两倍核转录组簇。该结果表明存在一种形式的“核法典” 多核细胞的转录组是通过两个或多个核簇“编码”的，结果是得到来自其他多核细胞群体的观察结果。这些编码的单元可以显示收缩力，抗凋亡的抵抗力甚至增殖潜力的差异，具有显着意义用于心脏功能和心力衰竭的发展。这个史蒂文·I。Katz提案的目的是1）使用尖端单细胞和单核测序识别和确认此核编码的技术，2）探索单声道和多种多样的空间分布 3）确定对遗传和环境的编码反应如何健康和失败的心脏扰动。根据该机制的要求，在此提案中，我们提出了一个计划为我们实验室的研究建立雄心勃勃的新方向，并以严格的工作为基础文学和合作者和主题专家的支持。该提案结合了生物信息学，成像和下一代测序方法，以确定核法典中的变化多核心肌细胞会导致对心脏损伤和压力的反应差异。完成后，该赠款将使对多核性和多倍体在哺乳动物的心脏，随后启示了许多新的诊断研究途径打击心力衰竭的治疗方法。