Understanding Cardiac Progenitors to deliver Regenerative Medicine and Disease Modelling

了解心脏祖细胞以提供再生医学和疾病建模

• 批准号: MR/T041668/1
• 负责人: Matthew Birket
• 金额: $ 113.02万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FT041668%2F1
• 关键词: Understanding; Cardiac; Progenitors; deliver; Regenerative

Heart disease, including damage to heart muscle, kills more people worldwide than any other illness. This means strategies for cardiac regeneration, such as cell replacement therapy after myocardial infarction and 'disease in a dish' modelling in vitro, are urgently needed. Access to cardiovascular cells including cardiomyocytes from human pluripotent stem cells (hPSCs) offers therapeutic potential capable of revolutionising cardiology but, as yet, the promise is unmet. Progress is currently hindered by our poor understanding of, and thus ability to control, cardiac cell development - principally the events of cardiac progenitor cell (CPC) specification, self-renewal and differentiation. The lack of comparative molecular benchmarking with human embryogenesis, required to authenticate cells in vitro, is another major barrier. By consequence, at present, mixtures of poorly specified cardiomyocyte identities emerge from uncontrolled, heterogeneous differentiation. The resultant cells are inadequate for accurate disease modelling (disease phenotypes are highly variable) and would be potentially life-threatening (arrhythmogenic) if transplanted. There are three major components to my study, delivery of which will unlock potential for academic research, clinical and industrial applications: 1) discovering the identity of CPCs and how to control them, 2) authenticating ventricular cardiomyocyte differentiation and 3) testing the system in an in vivo context.The challenge to unravelling the complexities of cardiac cell development is that differentiation is heterogeneous, CPCs are diverse, and cells progress along the differentiation trajectory at different rates. To overcome this, I have created an in vitro model system of single cell genomics linked to clonal lineage tracing and fate assessment. This integrated and unbiased approach will enable me to identify the gene expression and chromatin signatures pertinent to CPC self-renewal and cardiomyocyte differentiation potential. Perturbation experiments in vitro, combined with comparative measurements of the native cells in human embryogenesis, will help me identify the most functionally influential and authentic candidates. Together, these experiments will deliver the knowledge and tools to isolate, expand and differentiate the most desirable CPC identities - those with potential to be accurately programmed to specific target cell types e.g. ventricular cardiomyocytes (a key target for disease modelling and cell therapy). To learn how to accurately control downstream differentiation of CPCs to ventricular cardiomyocytes, I will use the gene expression and chromatin structure of human ventricular myocardium through development to benchmark differentiation in vitro. The relationship between signalling pathways and the regulation and expression of key transcription factors will be systematically resolved, to achieve the accurate, lineage-specific differentiation of CPCs. Finally, to validate these advances in CPC maintenance, tracking and differentiation to translational relevance, I will work in partnership with international collaborators to assess cell behaviour following transplantation into an animal model. In summary, by delivering research innovation of immediate clinical and industrial importance, this project is closely in tune with the UK Government's Industrial Strategy. The predicted advances will benefit applications in regenerative medicine, cardiac disease modelling, biomarker identification and drug screening. Moreover, it will also allow me to establish myself as a global leader in the field and, by partnering with industry, a persistent force towards achieving these translational goals.

心脏病，包括心肌损伤，在世界范围内比任何其他疾病都要多。这意味着迫切需要心脏再生的策略，例如心肌梗死后的细胞替代疗法和体外“培养皿中的疾病”模型。从人类多能干细胞（hPSC）中获得心血管细胞（包括心肌细胞）提供了能够彻底改变心脏病学的治疗潜力，但迄今为止，这一承诺尚未得到满足。目前，我们对心脏细胞发育的理解不足，因此控制心脏细胞发育的能力不足，主要是心脏祖细胞（CPC）的特化、自我更新和分化。另一个主要障碍是缺乏体外鉴定细胞所需的人类胚胎发生的比较分子基准。因此，目前，混合物的不良指定心肌细胞身份出现不受控制的，异质分化。所得到的细胞不足以进行准确的疾病建模（疾病表型是高度可变的），并且如果移植，将可能危及生命（致肿瘤）。我的研究有三个主要组成部分，其交付将释放学术研究，临床和工业应用的潜力：1）发现CPC的身份以及如何控制它们，2）验证心室心肌细胞分化，以及3）在体内环境中测试该系统。解开心脏细胞发育复杂性的挑战在于分化是异质的，CPC是多样的，并且细胞以不同的速率沿着分化轨迹前进。为了克服这一点，我已经创建了一个体外模型系统的单细胞基因组学与克隆谱系追踪和命运评估。这种综合和公正的方法将使我能够确定相关的CPC自我更新和心肌细胞分化潜力的基因表达和染色质签名。体外扰动实验，结合对人类胚胎发生中天然细胞的比较测量，将帮助我确定功能上最有影响力和最真实的候选者。总之，这些实验将提供知识和工具来分离，扩展和区分最理想的CPC身份-那些有可能被准确编程为特定靶细胞类型的CPC身份，例如心室心肌细胞（疾病建模和细胞治疗的关键目标）。 为了了解如何准确地控制CPC向心室心肌细胞的下游分化，我将通过发育使用人心室心肌的基因表达和染色质结构来基准体外分化。信号通路与关键转录因子的调控和表达之间的关系将被系统地解决，以实现CPC的准确的、谱系特异性的分化。最后，为了验证CPC维持，跟踪和分化与翻译相关性的这些进展，我将与国际合作者合作，评估移植到动物模型后的细胞行为。总之，通过提供具有直接临床和工业重要性的研究创新，该项目与英国政府的产业战略密切相关。预测的进展将有利于再生医学，心脏病建模，生物标志物鉴定和药物筛选的应用。此外，它还将使我成为该领域的全球领导者，并通过与行业合作，成为实现这些转化目标的持久力量。

项目成果

The generation and validation of two NKX2-5 fluorescent reporter human embryonic stem cell lines: UMANe002-A-1 and UMANe002-A-2

两种 NKX2-5 荧光报告基因人胚胎干细胞系 UMANe002-A-1 和 UMANe002-A-2 的生成和验证

• DOI: 10.1016/j.scr.2023.103262
• 发表时间: 2024
• 期刊: Stem Cell Research
• 影响因子: 1.2
• 作者: Douglas M

The generation and validation of a dual cardiac HAND1-Tomato NKX2-5-GFP human embryonic stem cell line UMANe002-A-3

双心脏 HAND1-Tomato NKX2-5-GFP 人胚胎干细胞系 UMANe002-A-3 的生成和验证

• DOI: 10.1016/j.scr.2024.103342
• 发表时间: 2024
• 期刊: Stem Cell Research
• 影响因子: 1.2
• 作者: Lynch A