Stem cell toxicology assays for cardiac differentiation

心脏分化的干细胞毒理学测定

• 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
• 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

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.

先天性心脏缺陷是最常见的出生缺陷之一 和胎儿心脏缺陷可能导致流产，终止高达 25%的孕妇环境因素可能影响高达80%的 CCD，但很少有人知道如何从环境中的化学物质可能会影响 心脏的形成。我们建议开发心源性（心脏的形成） 心脏）试验，利用干细胞来筛选化学物质，这重演了早期事件 在心脏形成中，心肌细胞、血管内皮细胞和 中胚层多能心肌细胞分化为血管平滑肌细胞 前体该测定在384孔板中进行，并使用鼠 胚胎干细胞（mESC）与报告基因的生物工程，以报告 心肌细胞、血管内皮细胞和血管平滑肌细胞的比例 从文化中涌现出来。在初步实验中，550种已知化学物质 用心肌生成试验筛选通路调节剂， 发现Wnt途径的表达强烈上调心脏 肌细胞这与Wnt途径对细胞凋亡的已知影响是一致的。 心脏发育和Wnt活性失调在 生产CCD。第一阶段的目标是通过以下方式进一步改进检测方法： 鉴定可以作为可靠的阳性鉴定的化合物或基因组构建体， 或系统中的负面效应，以开发方法，以最大限度地提高 可以从每次筛选运行中收集的信息， 方法，以提高一致性，并进一步简化测定。