Stem cell toxicology assays for cardiac differentiation

心脏分化的干细胞毒理学测定

基本信息

批准号：
8620564
负责人：
PATRICK M MCDONOUGH
金额：
$ 22.45万
依托单位：
VALA SCIENCES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-19 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8620564
关键词：
Affect Algorithms Appearance Area Biological Assay Biological Markers Biological Models Biomedical Engineering Blood Vessels Cardiac Cardiac Myocytes Cell Count Cell Differentiation process Cell Line Cells Chemicals Collaborations Congenital Abnormality Congenital Heart Defects Data Development Drug toxicity Embryo Embryology Embryonic Development Endothelial Cells Engineering Environment Environmental Health Environmental Risk Factor Event Genomics Goals Government Agencies Health Services Heart Human Incidence Institutes Live Birth Medical Research Mesoderm Methods MicroRNAs Mus Mutation Myocardium National Institute of Environmental Health Sciences Pathway interactions Phase Poison Pregnancy Production Proteins Provider Reporter Genes Reporting Research Research Institute Role Running Science Small Business Innovation Research Grant Smooth Muscle Smooth Muscle Myocytes Source Spontaneous abortion Stem cells System Testing Toxic Environmental Substances Toxic effect Toxicity Tests Toxicology Variant Vascular Endothelial Cell Vascular Endothelium Vascular Smooth Muscle base cardiogenesis cell type design drug discovery embryonic stem cell environmental chemical fetal heart cell heart function human embryonic stem cell human stem cells improved in vitro testing inhibitor/antagonist miniaturize novel programs public health relevance research study response screening standard care toxicant

项目摘要

Congenital cardiac defects (CCDs) are among the most common birth defects and fetal cardiac defects likely contribute to miscarriages, which terminate up to 25% of all pregnancies. Environmental factors may influence up to 80% of all CCDs, but little is known about how chemicals from the environment may affect formation of the heart. We propose to develop a cardiopoiesis (formation of the heart) assay to screen chemicals using stem cells, which recapitulate early event in heart formation, in which cardiac myocytes, vascular endothelial cells, and vascular smooth muscle cells differentiate from mesodermal multipotent cardiac precursors. The assay is performed in 384 well plates and uses murine embryonic stem cells (mESCs) bioengineered with reporter genes to report the proportion of cardiac myocytes, vascular endothelial cells, and vascular smooth that emerge from the cultures. In preliminary experiments, 550 known chemical pathway modulators were screened with the cardiopoiesis assay, and inhibitors of the Wnt pathway were found to strongly upregulate production of cardiac myocytes. This is consistent with the known influence of the Wnt pathway on cardiac development and the suspected role of dysregulated Wnt activity in producing CCDs. Phase I goals will be to further improve the assay by identifying compounds or genomic constructs that can serve as reliable positive or negative effectors in the system, to develop methods to maximize the information that can be garnered from each screening run, and to develop methods to improve the consistency and further miniaturize the assay.

先天性心脏缺陷（CCD）是最常见的出生缺陷之一胎儿心脏缺陷可能会导致流产，流产终止于占所有怀孕的 25%。环境因素可能影响高达 80% CCD，但人们对环境中的化学物质如何影响知之甚少心脏的形成。我们建议发展心脏生成（形成心脏）利用干细胞筛选化学物质的试验，重现早期事件在心脏形成过程中，心肌细胞、血管内皮细胞和血管平滑肌细胞从中胚层多能心肌细胞分化前体。该测定在 384 孔板中进行，并使用小鼠使用报告基因进行生物工程改造的胚胎干细胞（mESC）可报告心肌细胞、血管内皮细胞、血管平滑肌细胞比例从文化中产生的。在初步实验中，550 种已知化学物质通过心肌生成测定筛选途径调节剂，并通过抑制剂筛选途径调节剂发现 Wnt 通路的一部分可以强烈上调心肌细胞的产生肌细胞。这与 Wnt 通路对心脏发育和 Wnt 活性失调的可疑作用生产 CCD。第一阶段的目标是进一步改进检测方法鉴定可作为可靠阳性的化合物或基因组构建体或系统中的负面效应器，以开发方法来最大化可以从每次筛选运行中收集的信息，并开发提高一致性并进一步小型化测定的方法。