Rejuvenating Aging Human Cells through Transcriptional Reprogramming

通过转录重编程使衰老的人类细胞恢复活力

• 批准号: 9978411
• 负责人: HAO LI
• 金额: $ 28.23万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978411
• 关键词: Age; Aging; Back; Bioinformatics; Biological Models; CRISPR/Cas technology; Cardiovascular Diseases; Cell Culture Techniques; Cell Differentiation process; Cells; Chronic Disease; Disease; Disease model; Dreams; Drug Combinations; Fibroblasts; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genetic Transcription; Health; Human; Imagination; Individual; Malignant Neoplasms; Medical; Methodology; Modeling; Molecular; Morphology; Neurodegenerative Disorders; Organ; Pharmaceutical Preparations; Phenotype; Plant Roots; Population; Prevention; Rejuvenation; Reporter; Research; Reverse Transcription; Risk Factors; Symptoms; Technology; Testing; Therapeutic; Time; Tissue Therapy; Youth; age related; artist; base; combinatorial; fictional works; high throughput screening; human tissue; induced pluripotent stem cell; molecular phenotype; new technology; novel; novel strategies; novel therapeutics; programs; single-cell RNA sequencing; small molecule; stem cell differentiation; success; transcription factor

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.

衰老是许多慢性疾病的主要危险因素，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，与年龄有关的疾病 已经成为世界上最突出的健康问题之一。因此，现有的医疗实践 远集中在治疗每一种疾病，因为它出现独立。通过这种传统的方法， 疾病正迅速变得无法集体管理，因为为个人减轻一种疾病并不 阻止另一个不可避免的崛起。治疗和预防模式的根本转变将 要针对老化的根本原因，而不是其“症状”。在这种新的范式下， 人体组织和器官代表了一个有希望的和全新的方向。在这里，我们提出一个 采用非常规方法来应对我们这个时代最重大的挑战之一-系统地识别 复兴战略。 我们的建议是基于一个新的概念：通过转录恢复衰老的人类细胞 重新编程这是受到山中伸弥成功地从人类胚胎中提取诱导多能干细胞的启发。 通过转录重编程分化细胞。最基本的假设是， 和“老”是由特定基因表达程序定义的细胞的不同状态， 通过将转录因子（TF）的适当组合导入老细胞中，可以直接使衰老细胞再生。 使衰老的细胞恢复到年轻的状态 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， 该技术结合了CRISPR技术用于靶向基因调控，单细胞RNA测序用于基因调控， 表达谱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，我们将系统地鉴定TF的组合，当将其引入老年细胞时， 将老细胞的基因表达程序逆转为年轻细胞的。积极的打击从高 通量筛选之后将进行详细的细胞和分子表型分析，以确认 复兴前程的鉴定能够恢复活力的TF组合将为以下方面奠定独特的基础： 发现可以达到相同效果的小分子药物组合。药物激活每一个 可以使用转录报告基因筛选单个TF，并且可以对不同TF进行阳性命中 加起来这减少了寻找细胞再生的组合药物的不可能问题， 寻找激活特定TF的单一药物的问题要容易得多。