Molecular pathogenesis of congenital heart disease mediated by neural crest and second heart field cells

神经嵴和第二心野细胞介导先天性心脏病的分子发病机制

基本信息

批准号：
10621288
负责人：
BERNICE E MORROW
金额：
$ 61.84万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-12 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621288
关键词：
Actins Affect Aorta Arteries Branchial arch structure Cardiac Cell Communication Cell Lineage Cell Maturation Cell physiology Cells Cellular biology Chromosome Mapping Data Defect Development DiGeorge Syndrome Distal Embryo Embryonic Development Failure Future GATA3 gene Gene Expression Gene Mutation Gene Silencing Genes Genetic Genomics Heart Heart Abnormalities Human Immunofluorescence Immunologic In Situ Individual Knowledge Mediating Mesoderm Cell Molecular Morphogenesis Morphology Mus Muscle satellite cell Mutation Neural Crest Neural Crest Cell Neural tube Pathogenesis Patients Persistent Truncus Arteriosus Pharyngeal Apparatus Population Predisposition Process Risk Factors Role Signal Transduction Smooth Muscle Smooth Muscle Myocytes Specific qualifier value Structure Technology Testing Tissues Translating Vascular Smooth Muscle aortic arch cardiogenesis cell motility cell type conditional mutant congenital heart disorder gene discovery gene function gene regulatory network heart function in vivo interest migration mouse model mutant progenitor programs single-cell RNA sequencing stem cells whole genome

项目摘要

ABSTRACT The cardiac outflow tract (OFT) with adjoining major arteries is susceptible to developmental insults during embryogenesis that leads to congenital heart disease. Part of the reason for their vulnerability is because the morphogenesis of the OFT requires the interaction of both neural crest cells (NCCs) and adjacent second heart field (SHF) mesoderm cells. This takes place during dynamic expansion of the embryonic pharyngeal apparatus. To understand these two juxtaposed populations, we performed single cell RNA-sequencing (scRNA-seq) after lineage purification, focusing mainly on NCCs. Cardiac NCCs (CNCCs) have traditionally been defined positionally rather than molecularly by specific gene markers of cell fate progression because of their multipotency and changing genetic profiles. Using scRNA-seq, we were able to identify putative CNCCs in the pharyngeal apparatus. This was achieved by using expression of early vascular smooth muscle genes, such as Acta2, as a guide. From this, we identified three CNCC populations at mouse embryonic day, E10.5. We refer to them as the Tbx2 and Tbx3 (Tbx2/3), Isl1 and Acta2 populations, based upon distinct expression of these genes. We suggest that the Tbx2/3 and Isl1 lineage cells may independently evolve and contribute to smooth muscle cells of the pharyngeal arch arteries and OFT, respectively. This proposal is set to explore the origin and fate trajectories of these three populations by dual lineage tracing, by determining the function of genes within, and to build gene regulatory networks controlling their development. Not only will we examine changes in NCCs but also will evaluate surrounding SHF mesoderm cells because altering CNCCs might affect these progenitor cells as well. We also have scRNA-seq data on the SHF mesoderm cells purified from mouse embryos. On the other hand, it is possible that alteration of genes required in the SHF can disrupt CNCC development. As one specific example, Tbx1, the gene for 22q11.2 deletion syndrome is expressed in the SHF but not CNCCs, but it greatly affects their function. We will test the idea that in Tbx1 null mutant embryos, CNCCs fail to progress from Sox10 expressing progenitors and are not able to enter the OFT. Many of the genes to be investigated in this program are associated with congenital heart disease in human patients. Overall, this program will help identify molecular aspects of CNCCs and the orchestration of CNCC-SHF cell fates in embryogenesis.

抽象的邻近主要动脉的心脏流出道 (OFT) 很容易受到发育的影响胚胎发生过程中的损伤会导致先天性心脏病。他们的部分原因是脆弱性是因为 OFT 的形态发生需要两个神经元的相互作用嵴细胞 (NCC) 和邻近的第二心区 (SHF) 中胚层细胞。这发生了胚胎咽器动态扩张期间。要了解这两个并列群体，我们在谱系后进行单细胞 RNA 测序 (scRNA-seq) 纯化，主要集中于NCC。心脏 NCC（CNCC）传统上被定义为由于细胞命运进程的特定基因标记在位置上而不是在分子上他们的多能性和不断变化的遗传特征。使用 scRNA-seq，我们能够识别假定的 CNCC 位于咽部。这是通过使用早期的表达来实现的血管平滑肌基因，如Acta2，作为指导。由此我们确定了三小鼠胚胎日 E10.5 的 CNCC 群体。我们将它们称为 Tbx2 和 Tbx3 (Tbx2/3)、Isl1 和 Acta2 群体，基于这些基因的不同表达。我们表明 Tbx2/3 和 Isl1 谱系细胞可能独立进化并有助于分别为咽弓动脉和 OFT 的平滑肌细胞。本提案已定通过双谱系追踪来探索这三个人群的起源和命运轨迹确定基因的功能，并建立控制其基因的基因调控网络发展。我们不仅会检查 NCC 的变化，还会评估周围环境 SHF 中胚层细胞，因为改变 CNCC 也可能影响这些祖细胞。我们还有从小鼠胚胎中纯化的 SHF 中胚层细胞的 scRNA-seq 数据。上另一方面，SHF 所需基因的改变可能会破坏 CNCC 发展。作为一个具体的例子，Tbx1，22q11.2 缺失综合征的基因是在 SHF 中表达，但在 CNCC 中不表达，但它极大地影响了它们的功能。我们将测试这个想法在 Tbx1 无效突变胚胎中，CNCC 无法从表达 Sox10 的祖细胞发展而来并且无法进入 OFT。该计划中要研究的许多基因是与人类患者的先天性心脏病有关。总体而言，该计划将有助于识别 CNCC 的分子方面以及 CNCC-SHF 细胞命运的编排胚胎发生。