Rejuvenating Aging Human Cells through Transcriptional Reprogramming

老龄化是许多慢性疾病的主要风险因素，如心血管疾病， 神经退行性疾病和癌症。随着全球人口逐渐老龄化，与年龄相关的疾病 已经成为世界上最突出的健康问题之一。既有的医疗实践因此 FAR专注于在疾病出现时独立治疗每一种疾病。在这种传统的方法下，与年龄相关的 疾病正迅速变得集体难以控制，因为为个人减轻一种疾病并不能 防止不可避免的另一个国家的崛起。治疗和预防模式的根本性转变将 要针对衰老的根本原因，而不是它的“症状”。在这一新的范式下，老龄化的复兴 人体组织和器官代表了一个有希望的、根本上的新方向。在这里，我们提出一种 采用非常规方法来应对我们时代最重大的挑战之一--系统地确定 振兴战略。 我们的建议是基于一个新的概念：通过转录使衰老的人类细胞恢复活力 重新编程。这是受到山中成功地从 通过转录重编程而分化的细胞。基本的假设是，无论是“年轻”还是“年轻” 和“老”是细胞的不同状态，由特定的基因表达程序定义，并通过引入 将转录因子(TF)适当地结合到旧细胞中，就有可能直接使 并将它们带回年轻的状态。 为了验证这一假设并确定这样的转录程序，我们将开发一种新的方法 它结合了CRISPR技术用于靶向基因调控，单细胞RNA测序用于基因 表情分析和最先进的生物信息学分析。使用已建立的人类细胞培养模型 对于衰老研究，我们将系统地确定当被引入旧细胞时，能够 将衰老细胞的基因表达程序逆转为年轻细胞的基因表达程序。从高处打出积极的击球 在吞吐量筛选之后，将进行详细的细胞和分子表型分析，以确认 年轻化。确定能够恢复活力的TF组合将为以下工作奠定独特的基础 发现小分子药物的组合可以达到同样的效果。激活每一种药物的药物 可以使用转录报告对单个TF进行筛选，并且可以对不同TF进行阳性匹配 加在一起。这将搜索用于细胞恢复活力的组合药物的不可能问题减少到 更容易处理的问题是寻找一种能激活特定转铁蛋白的单一药物。

Aging is a major risk factor for many chronic diseases such as cardiovascular disease, neurodegenerative disease, and cancer. As the global population progressively ages, age-related diseases have become some of the world’s most prominent health problems. The established medical practice has thus far focused on treating each disease independently as it arises. With this conventional approach, age-related diseases are quickly becoming unmanageable collectively, as alleviating one disease for an individual does not prevent the inevitable rise of another one. A fundamental shift of paradigm in treatment and prevention would be to target the root cause of aging, instead of its “symptoms”. Under this new paradigm, rejuvenation of aging human tissues and organs represents a promising and fundamentally new direction. Here we propose an unconventional approach to tackle one of the grandest challenges of our time – to systematically identify strategies for rejuvenation. Our proposal is based on a novel concept: rejuvenating aging human cells through transcriptional reprogramming. This was inspired by Yamanaka’s success in deriving induced pluripotent stem cells from differentiated cells through transcriptional reprogramming. The fundamental hypothesis is that both “young” and “old” are different states of the cell defined by specific gene expression programs, and that by introducing appropriate combinations of transcription factors (TFs) into the old cells, it is possible to directly rejuvenate the old cells and bring them back to the youthful state. To test this hypothesis and to identify such transcriptional programs, we will develop a novel approach that combines the CRISPR technology for targeted gene regulation, single cell RNA sequencing for gene expression profiling, and state-of-the-art bioinformatic analysis. Using established human cell culture models for aging study, we will systematically identify combinations of TFs that, when introduced to old cells, are able to reverse the gene expression program of the old cells to that of the young cells. Positive hits from the high throughput screening will be followed by detailed cellular and molecular phenotyping to confirm the effect of rejuvenation. The identification of combinations of TFs capable of rejuvenation will set a unique stage for discovering combinations of small molecule drugs that can achieve the same effect. Drugs that activate each individual TF can be screened using transcriptional reporters, and positive hits for different TFs can be combined. This reduce the impossible problem of searching for combination drugs for cell rejuvenation to a much more manageable problem of searching for a single drug that activates a specific TF.