Unipotency of Tbx5+ embryonic cardiac progenitor cells to cardiomyocytes.

Tbx5 胚胎心脏祖细胞对心肌细胞的单能性。

• 批准号: MR/J007625/1
• 负责人: Kenta Yashiro
• 金额: $ 41万
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FJ007625%2F1
• 关键词: Unipotency; Tbx5+; embryonic; cardiac; progenitor

Heart failure is a major cause of death in industrialized countries. Although the present medicines can attenuate the symptoms to some extent, the expected outcome is still poor. Therefore a new treatment is needed as soon as possible. Since heart failure is caused by the loss or poor functioning of heart muscle cells, destroyed or damaged during a heart attack, regeneration therapy seems promising by the replenishment of the damaged heart muscle with new heart muscle cells or heart muscle stem cells (cardiac progenitor cells; CPCs) created in the laboratory. However our current knowledge is too limited to be able to create sufficiently safe and effective cells for clinical use. Thus a much better understanding of CPCs is critical to bring such regeneration therapy into the clinical arena.It is known that CPCs are composed of two different populations known as the first and the second heart field. The first and the second heart fields are the origin of left side and the right side of the heart, respectively. A factor called Tbx5 is present in the first heart field and another two factors called Nkx2-5 and Isl1 are present in both the first and the second heart fields. CPCs positive for Nkx2-5 or Isl1 are known to give rise to heart muscle cells as well as other components of the heart. On the other hand, although CPCs positive for Tbx5, the first heart field, are known to produce heart muscle cells, whether the first heart field can also produce other cell types present in the heart is still largely unknown.To better understand CPCs, we have been studying CPCs from the earliest stage during embryogenesis with a state-of-art technique, single cell cDNA analysis. This technique allows us to study each a single CPC one by one, which brings detailed information with a resolution that other methods have never been able to provide. According to our data, we found the novel evidence that the first heart field CPCs are initially positive only for Tbx5, then become triple positive for Nkx2-5, Tbx5 and Isl1, and subsequently become double positive for Nkx2-5 and Tbx5. To confirm conclusively this sequential change of the first heart field CPCs according to the temporal order, we initially tried to trace Tbx5-expressing CPCs in the mouse embryonic heart by deliberately labeling Tbx5-expressing CPCs permanently with a fluorescent protein, eYFP. From this pilot study, we assume that the first heart field of Tbx5-expressing CPCs produces only heart muscle cells, which is quite different from Nkx2-5-expressing or Isl1-expressing CPCs.In this project, we will confirm whether our original assumption that Tbx5-expressing CPCs produce only heart muscle cells is true or not. We will trace Tbx5-expressing CPCs in detail with permanent eYFP label. This will enable us to consolidate the evidence of the sequential change of Tbx5-expressing CPCs as well as to clearly identify what type of the cells Tbx5-expressing CPCs give rise to. Furthermore, we aim to establish a new method to study Tbx5-expressing cardiac progenitor cells derived from mouse embryonic stem cells. Using this new method, we can further study Tbx5-expressing CPCs in vitro without using of animals, and we can test the use of Tbx5-expressing CPCs for the treatment of animal models of heart failure by the efficient enrichment of Tbx5-expressing CPCs. Thus, successful completion of this project will clearly demonstrate whether Tbx5-expressing CPCs can only provide heart muscle cells. Such novel knowledge in basic science as well as the establishment of a useful new tool for basic science study will greatly improve our understanding of heart muscle development. We strongly believe that completion of this project is one of the many steps towards the overall goal of the establishment of future regeneration therapies for heart failure.

心力衰竭是工业化国家死亡的主要原因。尽管目前的药物可以在某种程度上减轻症状，但预期的结果仍然很差。因此，尽快需要新的治疗方法。由于心力衰竭是由心脏肌肉细胞的丧失或功能不佳引起的，在心脏病发作过程中破坏或受损，因此再生疗法似乎有望通过在实验室中形成的新心肌细胞或心脏肌肉干细胞（心脏祖细胞； CPC）补充受损的心肌。但是，我们目前的知识太限制了，无法创建足够安全有效的细胞供临床使用。因此，对CPC的更好理解对于将这种再生疗法带入临床领域至关重要。众所周知，CPC由两个不同的人群组成，称为第一个和第二个心脏场。第一个和第二个心脏场是左侧和心脏右侧的起源。第一个心脏场中存在一个称为TBX5的因素，另外两个称为NKX2-5和ISL1的因素都存在于第一个和第二个心脏场中。已知NKX2-5或ISL1阳性的CPC会产生心脏肌肉细胞以及心脏的其他成分。另一方面，尽管众所周知，tbx5呈阳性的CPC呈阳性，它会产生心脏肌肉细胞，但第一个心脏领域是否还能产生心脏中存在的其他细胞类型，但仍未知。为了更好地了解CPC，我们一直在研究CPC从胚胎中研究CPC在胚胎生成过程中，从而通过既有前部技术，单个细胞cdna分析，，单细胞cdna分析。这项技术使我们能够一一研究每个CPC，从而带有其他方法无法提供的分辨率的详细信息。根据我们的数据，我们发现了新的证据表明，第一个心脏场CPC最初仅对TBX5呈阳性，然后在NKX2-5，TBX5和ISL1中变为三重阳性，随后在NKX2-5和TBX5中成为双重阳性。为了根据时间顺序结束对第一心脏场CPC的顺序变化，我们最初试图通过故意用荧光蛋白EYFP永久标记表达TBX5 CPC的小鼠胚胎心脏中表达TBX5的CPC。从这项试验研究中，我们假设表达TBX5的CPC的第一个心脏场仅产生心脏肌肉细胞，这与表达NKX2-5表达或表达ISL1的CPCS。我们将详细介绍具有永久EYFP标签的详细表达TBX5的CPC。这将使我们能够巩固表达TBX5的CPC的顺序变化的证据，并清楚地识别出表达TBX5 CPC的细胞类型的类型。此外，我们旨在建立一种研究源自小鼠胚胎干细胞的表达TBX5心脏祖细胞的新方法。使用这种新方法，我们可以在不使用动物的情况下进一步研究表达TBX5的CPC，并且可以通过有效地富集表达TBX5的CPC来测试使用TBX5表达CPC来治疗心力衰竭的动物模型。因此，该项目的成功完成将清楚地证明表达TBX5的CPC是否只能提供心肌细胞。这种新的基础科学知识以及建立有用的基础科学研究工具将大大提高我们对心肌发展的理解。我们坚信，该项目的完成是朝着建立未来再生疗法进行心力衰竭的整体目标的众多步骤之一。

项目成果

GFRA2 Identifies Cardiac Progenitors and Mediates Cardiomyocyte Differentiation in a RET-Independent Signaling Pathway.

• DOI: 10.1016/j.celrep.2016.06.050
• 发表时间: 2016-07-26
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Ishida H;Saba R;Kokkinopoulos I;Hashimoto M;Yamaguchi O;Nowotschin S;Shiraishi M;Ruchaya P;Miller D;Harmer S;Poliandri A;Kogaki S;Sakata Y;Dunkel L;Tinker A;Hadjantonakis AK;Sawa Y;Sasaki H;Ozono K;Suzuki K;Yashiro K
• 通讯作者: Yashiro K

Retinoic acid signaling regulates sonic hedgehog and bone morphogenetic protein signalings during genital tubercle development.

• DOI: 10.1002/bdrb.20344
• 发表时间: 2012-02
• 期刊: Birth defects research. Part B, Developmental and reproductive toxicology
• 作者: Liu L;Suzuki K;Nakagata N;Mihara K;Matsumaru D;Ogino Y;Yashiro K;Hamada H;Liu Z;Evans SM;Mendelsohn C;Yamada G
• 通讯作者: Yamada G