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

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

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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）是最常见的先天缺陷之一胎儿心脏缺陷可能导致流产，终止所有怀孕的25％。环境因素可能会影响多达80％ CCD，但对环境的化学物质如何影响很少知道心脏的形成。我们建议开发一种心脏（形成心脏）使用干细胞进行筛选化学物质的测定，这些干细胞概括了早期事件在心脏形成中，心肌细胞，血管内皮细胞和血管平滑肌细胞与中胚层多胎心脏区分开前体。该测定在384个井板中进行，并使用鼠胚胎干细胞（MESC）用报告基因进行生物工程，以报告心肌细胞，血管内皮细胞和血管光滑的比例从文化中浮现出来。在初步实验中，有550种已知化学物质用心poiesis分析和抑制剂筛选途径调节剂发现Wnt途径的强烈上调心脏的产生心肌细胞。这与Wnt途径对已知的影响一致心脏发展和WNT活性失调的可疑作用在生产CCD。第一阶段目标是进一步改善测定法识别可以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子，开发方法以最大化可以从每次筛选中获得的信息，并开发提高一致性并进一步微型化测定法的方法。