Phenotypic Screening for Longevity Interventions Using Single-cell Epigenetic Signatures

使用单细胞表观遗传特征进行长寿干预的表型筛选

• 批准号: 10225468
• 负责人: ALEXEY V TERSKIKH
• 金额: $ 24.38万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10225468
• 关键词: Age; Aging; Animals; Antibodies; Apoptosis; Aspirin; Biological Assay; Biology of Aging; Blood Cells; C57BL/6 Mouse; Cell Count; Cell Nucleus; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Custom; Designer Drugs; Disease; Dose; Drug Prospecting; Drug Screening; Drug Targeting; Epigenetic Process; Flowcharts; Genome; Glioblastoma; Hepatocyte; Heterochromatin; Histones; Human; Image; In Vitro; Inbreeding; Incidence; Intervention; Knowledge; Libraries; Longevity; Machine Learning; Mediating; Microscopy; Modeling; Molecular; Mus; Nature; Normal Cell; Nuclear; Pattern; Pharmaceutical Preparations; Phenotype; Plant Roots; Powder dose form; Rejuvenation; Sirolimus; Specificity; Splenocyte; Stains; Sum; Techniques; Testing; Toxic effect; Validation; Yeasts; base; cost; deep sequencing; drug candidate; drug testing; epigenetic profiling; epigenome; healthy aging; histone methylation; in vivo; microscopic imaging; neoplastic cell; novel; novel therapeutics; research and development; response; screening; side effect; small molecule libraries; tool

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.

项目摘要 对衰老生物学理解的最新进展提高了药物治疗的前景。 促进人类健康老龄化的干预措施。然而，这种干预措施应 基本上没有毒性或副作用。在实践中，药物检测的吞吐量（目前 在动物中进行）是确定干预措施的主要限制之一， 健康老龄化因此，一个挑战是确定能够诱导返老还童的候选药物 和/或健康的老化，以高通量的方式具有最小的副作用。在此，我们建议 利用基于表观遗传成像的新的高内容筛选方法， 单细胞中的景观。为了迎接挑战，我们建议利用一本小说 我们已经开发的技术，是植根于表观基因组地形分析在 单细胞水平、自动显微镜和机器学习。“显微成像 表观遗传景观”（MIEL）捕获表观遗传标记的核染色模式（例如， 乙酰化和甲基化组蛋白），并采用机器学习来准确区分 在这些模式之间。针对这一应用，我们已经测试了MIEL方法是否 适用于在细胞水平上建立基于表型细胞的长寿诱导物测定。 我们的初步结果验证了MIEL测定用于高含量筛选应用以鉴定 新的候选促长寿化合物。总之，我们已经开发并验证了一个高 能够鉴定促长寿/健康衰老的基于内容物细胞的筛选测定 化合物基于其对表观遗传特征的影响。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。验证 细胞再生的新诱导剂。