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

• 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
• 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

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.

项目概要 心力衰竭抵抗了其他疾病死亡率下降的趋势，新疗法正在出现 需要。心力衰竭是由心肌细胞肥大和损失引起的，心肌细胞构成了心脏的大部分。 心脏的容量并提供将血液输送到全身所需的主要收缩力。超过 哺乳动物中 90% 的心肌细胞是多倍体和/或多核的。这些额外的副本 DNA 已在心脏再生能力较低的背景下进行了研究，但相对较少 人们的努力集中在这种增加的核性所带来的其他潜在的适应性或适应不良影响上。 从历史上看，成熟心肌细胞的大尺寸阻碍了使用无偏单细胞对其进行分析 RNA测序。研究重点是孤立核中的 RNAseq，其本质上缺乏 有关所研究心脏内核的信息。最近，新技术使得公正、 完整成熟心肌细胞的高通量单细胞 RNAseq。当这些单细胞的结果 将研究与孤立核研究进行比较，研究人员发现，使用相同的聚类 算法中，不同细胞转录组簇的数量大约是不同细胞转录组簇数量的两倍 核转录组簇。这一结果表明存在一种“核代码”形式，其中 多核细胞的转录组是由两个或多个核簇中的核“编码”的，结果 得到其他多核细胞群观察结果的支持。这些编码单元可以显示 收缩性、细胞凋亡抵抗力甚至增殖潜力的差异，具有重大影响 用于心脏功能和心力衰竭的发展。 Steven I. Katz 提案的目标是 1) 利用尖端的单细胞和单核测序 技术来识别和确认这种核编码，2）探索单核和多核的空间分布 心脏中的有核细胞，以及 3) 确定这种编码如何响应遗传和环境 健康和衰竭心脏的干扰。根据该机制的要求，我们在本提案中提出了 为我们实验室的研究计划一个雄心勃勃的新方向，建立在严格的工作记录的基础上 文献并得到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来确定核密码的变化 多核心肌细胞导致对心脏损伤和应激的反应存在差异。完成后， 这笔赠款将使人们更全面地了解多核和多倍体在 哺乳动物心脏，随后揭示了许多新的诊断和研究途径 对抗心力衰竭的治疗方法。