背景:在细胞水平上抑制或减缓衰老标志已被提出作为增加机体寿命和健康跨度的途径。因此,人们对抗衰老药物的发现非常感兴趣。然而,这目前需要费力和冗长的寿命分析。这里,我们提出了一种新的筛选读数,用于快速发现抑制体外细胞群老化并延长体内寿命的化合物。
方法:使用Illumina甲基化阵列,我们监测了随培养的成人原代细胞长期传代而发生的DNA甲基化变化.这使我们能够发展,test,并验证CellPopAge时钟,带有底层算法的表观遗传时钟,在现有的表观遗传时钟中,其设计用于检测体外抗衰老化合物。此外,我们测量了衰老的标志物,并在果蝇体内进行了长寿实验,进一步验证我们发现新型抗衰老化合物的方法。最后,我们将我们的表观遗传时钟与其他可用的表观遗传时钟进行基准标记,以巩固其对培养中原代细胞的有用性和专业化。
结果:我们开发了一种新的表观遗传时钟,CellPopAge时钟,准确监测成人原代细胞群的年龄。我们发现CellPopAgeClock可以检测用雷帕霉素或曲美替尼处理的人原代细胞的基于传代的老化速度,成熟的长寿药。然后,我们利用CellPopAge时钟作为筛选工具,用于鉴定减缓细胞群体衰老的化合物,发现新型抗衰老药物,torin2和dactolisib(BEZ-235)。我们证明,用抗衰老化合物处理的人类原代细胞的表观遗传衰老伴随着衰老和衰老生物标志物的减少。最后,我们通过利用特殊配方的碱性介质来增加果蝇的药物生物利用度,从而在体内扩展了我们的筛选平台。我们证明了新型抗衰老药物,torin2和dactolisib(BEZ-235),增加体内寿命。
结论:我们的方法将CpG甲基化分析的范围扩大到在体外使用人细胞准确、快速地检测药物的抗衰老潜力,在体内,提供了一个新颖的加速发现平台,以测试所寻求的抗衰老化合物和老虎机。
BACKGROUND: Restraining or slowing ageing hallmarks at the cellular level have been proposed as a route to increased organismal lifespan and healthspan. Consequently, there is great interest in anti-ageing drug discovery. However, this currently requires laborious and lengthy
longevity analysis. Here, we present a novel screening readout for the expedited discovery of compounds that restrain ageing of cell populations in vitro and enable extension of in vivo lifespan.
METHODS: Using Illumina methylation arrays, we monitored DNA methylation changes accompanying long-term passaging of adult primary human cells in culture. This enabled us to develop, test, and validate the CellPopAge Clock, an epigenetic clock with underlying algorithm, unique among existing epigenetic clocks for its design to detect anti-ageing compounds in vitro. Additionally, we measured markers of senescence and performed
longevity experiments in vivo in Drosophila, to further validate our approach to discover novel anti-ageing compounds. Finally, we bench mark our epigenetic clock with other available epigenetic clocks to consolidate its usefulness and specialisation for primary cells in culture.
RESULTS: We developed a novel epigenetic clock, the CellPopAge Clock, to accurately monitor the age of a population of adult human primary cells. We find that the CellPopAge Clock can detect decelerated passage-based ageing of human primary cells treated with rapamycin or trametinib, well-established
longevity drugs. We then utilise the CellPopAge Clock as a screening tool for the identification of compounds which decelerate ageing of cell populations, uncovering novel anti-ageing drugs, torin2 and dactolisib (BEZ-235). We demonstrate that delayed epigenetic ageing in human primary cells treated with anti-ageing compounds is accompanied by a reduction in senescence and ageing biomarkers. Finally, we extend our screening platform in vivo by taking advantage of a specially formulated holidic medium for increased drug bioavailability in Drosophila. We show that the novel anti-ageing drugs, torin2 and dactolisib (BEZ-235), increase
longevity in vivo.
CONCLUSIONS: Our method expands the scope of CpG methylation profiling to accurately and rapidly detecting anti-ageing potential of drugs using human cells in vitro, and in vivo, providing a novel accelerated discovery platform to test sought after anti-ageing compounds and geroprotectors.