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.

抽象的具有毗邻主要动脉的心脏流出道（通常）容易发育胚胎发生过程中导致先天性心脏病的侮辱。他们的部分原因脆弱性是因为OFT的形态发生需要两个神经的相互作用波峰细胞（NCC）和相邻的第二心脏场（SHF）中胚层细胞。这发生了在胚胎咽部装置的动态膨胀过程中。了解这两个并置的种群，我们在谱系后进行了单细胞RNA测序（SCRNA-SEQ）纯化，主要集中在NCC上。传统上定义了心脏NCC（CNCC）由于细胞命运进展的特定基因标记，而不是分子上它们的多履行和变化的遗传特征。使用scrna-seq，我们能够识别咽部设备中的推定CNCC。这是通过使用早期表达来实现的血管平滑肌基因，例如ACTA2，作为指南。从中，我们确定了三个小鼠胚胎日的CNCC种群，E10.5。我们称它们为TBX2和TBX3 （TBX2/3），ISL1和ACTA2种群，基于这些基因的独特表达。我们建议TBX2/3和ISL1谱系细胞可以独立发展并有助于咽弓动脉及其经常的平滑肌细胞。该建议已设定通过双重谱系跟踪，探索这三个人群的起源和命运轨迹确定基因的功能，并建立控制其基因调节网络发展。我们不仅会检查NCC的变化，而且还会评估周围 SHF中胚层细胞，因为改变CNCC可能也会影响这些祖细胞。我们还具有从小鼠胚胎纯化的SHF中胚层细胞上的SCRNA-SEQ数据。在另一方面，SHF中所需的基因的改变可能会破坏CNCC 发展。作为一个具体示例，TBX1，22Q11.2缺失综合征的基因是在SHF中表达，但不表达CNCC，但极大地影响了它们的功能。我们将测试这个想法在TBX1 NULL突变胚胎中，CNCC无法从SOX10表达祖细胞中进展并且无法进入。在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，该计划将有所帮助确定CNCC的分子方面和CNCC-SHF细胞命运的编排胚胎发生。