Cellular and Tissue Rejuvenation through Transcriptional Reprogramming

通过转录重编程实现细胞和组织的年轻化

基本信息

批准号：
10729260
负责人：
HAO LI
金额：
$ 61.82万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10729260
关键词：
Aging Back Behavioral Assay Blood Blood specimen Brain CRISPR/Cas technology Cell Culture Techniques Cells Chromatin Remodeling Factor Civilization Disease Disease model Dreams EZH2 gene Environment Fibroblasts Gene Expression Profiling Genetic Transcription Grant Histology Human Human body Imagination In Vitro Individual Intervention Liver Longevity Methodology Modeling Molecular Mus Organ Personal Satisfaction Pharmaceutical Preparations Phenotype Prevention Rejuvenation Repression Research STAT3 gene Sampling Technology Testing Tissues Transgenic Mice United States National Institutes of Health Youth age related aged artist cell age combinatorial comparative computerized tools fictional works high throughput screening human tissue improved in vivo molecular phenotype new technology novel strategies overexpression programs small molecule stem cells transcription factor transcriptional reprogramming transcriptome translation to humans

项目摘要

Summary Organismal and cell rejuvenation are exciting new approaches to counteract aging, and recent breakthroughs have brought them to the forefront of aging research. For examples, systemic factors in young blood was found to rejuvenate various mouse tissues and brain function, and partial reprogramming with four stem cell transcription factors (TFs) (Yamanaka factors) rejuvenate tissues and cells and extend the lifespan of mice. These discoveries demonstrate that “young” and “old” can be described as different states, and the “old” state can be reversed back into a “young” state through transcriptional reprogramming. We hypothesized that there might exist many solutions to human cell rejuvenation through transcriptional reprogramming, and some of the solutions may be safer and more potent than Yamanaka factors. In a recently completed project supported by an NIH/NIA R21 grant, the Li lab developed a systematic approach to test this hypothesis and to find the solutions. Using a human cell culture model of replicative aging employed by Hayflick (continuously passaged human fibroblast cells), we developed a high throughput screen using Perturb-seq to identify the potential rejuvenating TFs -- those that when over-expressed or repressed in old cells, are capable of reprogramming the global gene expression program from the old state back to a younger state. We identified four TFs/chromatin modifier (E2F3, EZH2, STAT3, ZFX) that when over-expressed or repressed individually, are able to rejuvenate human fibroblast cells aged in vitro. Here we propose to further test the rejuvenating effect of these four factors in human fibroblast cells aged in vivo in their natural tissue environment, and in mouse liver. We will also develop new technologies to screen for more potent rejuvenating TF combinations and test them in aged human fibroblast cells and in mouse liver. If successful, this proposed study will identify TFs/TF combinations that can rejuvenate in vitro and in vivo aged human fibroblast cells and mouse liver. This will set the stage for transgenic mouse study and translation to human therapies. The methodologies developed in this proposal can be generalized to identify combinatorial transcriptional programs that produce any desired cellular phenotypes, e.g., the reversion of the cellular state from disease to normal in cell culture models of diseases.

概括有机体和细胞复兴是对抗衰老的令人兴奋的新方法，也是最近的突破例如，在年轻血液中发现了系统性因素。使小鼠的各种组织和大脑功能恢复活力，并用四种干细胞进行部分重编程转录因子 (TF)（山中因子）可以使组织和细胞恢复活力并延长小鼠的寿命。这些发现表明，“年轻”和“年老”可以描述为不同的状态，而“年老”的状态可以通过转录重编程逆转回“年轻”状态。我们认为，通过转录可能存在多种人类细胞再生的解决方案重新编程，其中一些解决方案可能比最近的山中因子更安全、更有效。已完成由 NIH/NIA R21 拨款支持的项目，李实验室开发了一种系统方法来测试该项目假设并使用复制衰老的人类细胞培养模型找到解决方案。 Hayflick（连续传代的人成纤维细胞），我们使用 Perturb-seq 来识别潜在的恢复活力的转录因子——那些在旧的转录因子中过度表达或抑制的转录因子细胞能够将全局基因表达程序从旧状态重新编程回较年轻的状态我们确定了四种 TF/染色质修饰剂（E2F3、EZH2、STAT3、ZFX），当过度表达或单独抑制，能够使体外老化的人类成纤维细胞恢复活力。这里我们建议进一步测试这四个因素对老化的人成纤维细胞的返老还童作用。我们还将开发新技术来筛选。寻找更有效的恢复活力的 TF 组合，并在衰老的人类成纤维细胞和小鼠肝脏中进行测试。如果成功，这项拟议的研究将确定可以在体外和体内使衰老的人恢复活力的 TF/TF 组合人类成纤维细胞和小鼠肝脏这将为转基因小鼠的研究和转化奠定基础。该提案中开发的方法可以推广到识别组合疗法。产生任何所需细胞表型的转录程序，例如细胞状态的逆转在疾病的细胞培养模型中从疾病到正常。