Phenotypic Screening for Longevity Interventions Using Single-cell Epigenetic Signatures

项目总结 在了解衰老生物学方面的最新进展增加了药物的前景 促进人类健康老龄化的干预措施。然而，这种干预应该有 基本上没有毒性或副作用。在实践中，药物测试的吞吐量(目前 在动物中进行)是确定促进 健康的衰老。因此，一个挑战是确定能够诱导返老还童的候选药物。 和/或健康老化，以高通量方式最大限度地减少副作用。在此，我们建议 利用一种新的基于表观遗传成像的高含量筛选方法 单细胞中的风景。为了迎接挑战，我们建议利用一部小说 我们开发的技术植根于表观基因组图谱的分析 单细胞水平、自动显微镜和机器学习。“显微成像技术” 表观遗传景观“(MIEL)捕捉了表观遗传标记的核染色模式(例如 乙酰化组蛋白和甲基化组蛋白)，并使用机器学习来准确区分 在这样的模式之间。与此应用程序相关，我们测试了MIEL方法是否 适合于在细胞水平上建立基于表型细胞的长寿诱导剂的分析。 我们的初步结果验证了MIEL试验在高含量筛选应用中的有效性 新的候选长寿化合物。总而言之，我们开发并验证了一种高 基于内容细胞的筛选方法能够识别长寿/健康衰老 基于它们对表观遗传特征的影响的化合物。我们的具体目标如下： 具体目标1.筛选新的细胞再生诱导剂。具体目标2.确认 新型细胞复壮诱导剂。

PROJECT SUMMARY Recent advances in understanding the biology of aging have raised the prospect of drug interventions to promote healthy aging in humans. However, such interventions should have essentially no toxicity or side effects. In practice, the throughput of drug testing (currently conducted in animals) is one of the major limitations for identifying interventions to promote healthy aging. Hence, a challenge is to identify candidate drugs able to induce rejuvenation and/or healthy aging, with minimal side effects in a high throughput fashion. Here we propose to take advantage of a novel high content screening approach based on imaging of epigenetic landscape in single cells. To meet the challenge, we propose to take advantage of a novel technique we have developed that is rooted in the analysis of epigenome topography at the single cell level, automated microscopy, and machine learning. “Microscopic Imaging of Epigenetic Landscapes” (MIEL) captures patterns of nuclear staining of epigenetic marks (e.g. acetylated and methylated histones) and employs machine learning to accurately distinguish between such patterns. Pertinent to this application, we have tested whether MIEL approach is suitable for building a phenotypic cell-based assay for inducers of longevity at the cellular level. Our preliminary results validated MIEL assay for a high content screening application to identify novel candidate pro-longevity compounds. In sum, we have developed and validated a high content cell-based screening assay capable of identifying pro-longevity/healthy aging compounds based on their effect on epigenetic signature. Our specific Aims are as follows: Specific Aim 1. Screen for novel inducers of cellular rejuvenation. Specific Aim 2. Validation of novel inducers of cellular rejuvenation.