Integration of Genomic Biomarkers with the devTOX Human Embryonic Stem Cells Scre

描述（由申请方提供）：目前用于评估药物和环境因子导致出生缺陷的可能性的试验涉及使用动物模型。这些模型不仅成本高、耗时长，而且与人类数据的一致性也很差。Stemina TM Biomarker Discovery（Stemina）开发了一种筛选试验，该试验利用人胚胎干（hES）细胞和代谢组学，在全人类模型系统中研究暴露于供试化合物的hES细胞分泌组，以鉴定具有致畸潜力的化合物。集成实验室系统（ILS）和Stemina正式建立合作伙伴关系，以推进DevTOXTM，Stemina的基于干细胞的毒理学测试平台。该SBIR将深化和扩展独特的devTOXTM平台，通过询问毒物暴露对hES细胞的人类基因组和表观基因组的影响。关注对表观基因组的影响的毒性测试筛选是基于毒性的筛选的一个方面，除了少数例外，在基于高含量细胞的毒性测定中几乎不存在。重点 该SBIR的目的是：1.使用devTOXTM平台进行时间过程和剂量反应研究，旨在评价指示异常hES细胞功能的已知和研究性表观遗传生物标志物，以及2.将基于毒理基因组学的mRNA和miRNA生物标志物整合到devTOXTM平台，以评估毒物对基因组和表观基因组的影响，具体目标有三个。具体目的1是评估与在细胞培养扩增期间维持hES细胞的自我更新和多能性相关的生物标志物基因和miRNAS的稳定性。具体目标2评估测试化合物的参考组对hES细胞中的基因组和表观基因组应答生物标志物的影响。具体目标3将在DevTOXTM中测试由陶氏化学公司提供的10种盲态化学品，并整合基因组和表观基因组反应生物标志物，然后将结果与陶氏化学公司其他发育毒理学筛选的“内部”测试结果进行比较。我们预计，基因表达和表观基因组分析与devTOXTM的整合将提高devTOXTM平台的预测性，增加其作为预测潜在发育和生殖毒性的证据权重工具的使用。 公共卫生相关性：干细胞在人体所有细胞中具有独特的自我更新能力，也就是说，它们可以保持在原始的非特化状态。在适当的条件下，它们可以产生专门的身体细胞（分化），如心脏，肝脏或胰腺。人类胚胎干细胞正在被开发为评估发育毒性的毒性测试平台。这些细胞提供了一个独特的模型系统，以了解和评估环境因子和新候选药物的影响，以预测或预测人类的毒性。

DESCRIPTION (provided by applicant): Current assays employed to assess the potential of drugs and environmental agents for their potential to cause birth defects involve the use of animal models. Not only are these models costly and time consuming, but they also have poor concordance to human data. SteminaTM Biomarker Discovery (Stemina) has developed a screening assay that utilizes human embryonic stem (hES) cells and metabolomics to study the secretome of hES cells exposed to test compounds in an all human model system to identify compounds with teratogenic potential. Integrated Laboratory Systems (ILS) and Stemina have formalized a partnership to advance devTOXTM, Stemina's stem cell-based toxicology testing platform. This SBIR will deepen and extend the unique devTOXTM platform by interrogating impact on the human genome and epigenome of hES cells from toxicant exposures. Toxicity testing screens focused on effects to the epigenome are aspect of toxicity based screening that with a few exceptions is virtually absent in high content cellular based toxicity assays. The focus of this SBIR is to: 1. conduct time course and dose-response studies directed at evaluating known and investigative epigenetic biomarkers indicative of aberrant hES cell function using the devTOXTM platform and 2. integrate toxicogenomics-based mRNA and miRNA biomarkers to the devTOXTM platform to assess the impact of toxicants on the genome and epigenome in three specific aims. Specific Aim 1 is to assess stability of biomarker genes and miRNAS associated with maintaining self- renewal and pluripotency of hES cells during cell culture expansion. Specific aim 2 Assess the impact of a reference set of test compounds on genomic and epigenomic response biomarkers in hES cells. Specific aim 3 will test 10 blinded chemicals provided by DOW Chemical Company in DevTOXTM integrated with genomic and epigenomic response biomarkers, then compare results to Dow "in-house" test results from other developmental toxicology screens. We anticipate that the integration of the gene expression and epigenomic profiling with the devTOXTM will increase the predictivity of the devTOXTM platform increasing its use as a weight- of-evidence tool in predicting potential development and reproductive toxicity. PUBLIC HEALTH RELEVANCE: Stem cells have the unique ability among all of the cells of the human body of self- renewal, that is, they can remain in a primitive unspecialized state. Under the right conditions, they can give rise to specialized cells of the body (differentiation) like th heart, liver, or pancreas. Human embryonic stem cells are being developed as a toxicity testing platform for the assessment of developmental toxicity. These cells present a unique model system to understand and assess the effects of environmental agents and new drug candidates to predict or anticipate toxicity in humans.