Human Gene Knockdowns that May Extend Lifespan

描述(申请人提供)：FOXO转录因子延长动物的寿命并延缓与年龄相关的疾病，许多研究现已将FOXO3a DNA变异与人类的非凡长寿联系在一起。因此，现在似乎是时候寻找可能调节FOXO依赖或其他长寿途径的人类基因了。FOXO蛋白可以通过多种方式被激活以延长动物的寿命。例如，线虫FOXO可以延长寿命，响应胰岛素/IGF-1信号的减少，5-羟色胺信号的改变，以及AMP激酶活性的增加，热休克因子活性的增加，Lin-4 microRNA活性的增加，Jun激酶活性的增加和其他输入。因此，可能有许多基因扰动可以延长人类的健康寿命；其中一些扰动可能比其他更安全和有效。因为不可能对长寿的人类进行基因筛查，我们转而在人类细胞中进行基因筛查。我们的实验策略是基于观察到，到目前为止测试的所有FOXO依赖的延长寿命的途径(以及许多其他延长寿命的途径)都增加了对氧化应激的抵抗力。尽管氧化应激抵抗在延长寿命中的作用尚不清楚，但两者之间的相关性是 足够紧密，以至于在许多模式生物中，对氧化应激抗性的筛选已经产生了长寿的突变。因此，为了获得一组潜在的人类长寿基因，凯尼恩实验室在人类原代细胞系中进行了全基因组siRNA筛选，以检测其对氧化应激的抗性。这些命中的基因包括已知的线虫FOXO调节器，其他长寿蛋白的调节器，如TOR和NRF2，以及新基因。从这一组中，凯尼恩实验室将确定新的人类长寿和健康跨度基因的良好候选者。要做到这一点，他们将确定哪些基因敲除会触发其他与寿命相关的因素，如异种生物抵抗力或自噬。此外，他们将询问哪些基因敲除扰乱了FOXO3a、TOR或NRF2的活动。最后，为了将这些基因与长寿联系起来，他们将测试它们的能力 影响线虫寿命以及百岁老人家庭中线虫表达的变化。这一新的方法将确定新的潜在药物目标，以延长人类生命的年轻和富有成效的岁月，并延缓与年龄相关的疾病，如癌症、心脏病和/或神经退行性疾病。

DESCRIPTION (provided by applicant): FOXO transcription factors extend lifespan and delay age-related disease in animals, and many studies have now linked FOXO3A DNA variants to exceptional longevity in humans. Thus, the time seems right to look for human genes that are likely to regulate FOXO-dependent, or other, longevity pathways. FOXO proteins can be activated in many ways to extend animal lifespan. For example, C. elegans FOXO can promote longevity in response to reduced insulin/IGF-1 signaling, altered serotonin signaling, and elevated AMP kinase activity, elevated heat-shock factor activity, elevated lin-4 microRNA activity, elevated Jun kinase activity and other inputs. Thus, there could be many gene perturbations that can extend healthy lifespan in humans; and some of these perturbations may be safer and more effective than others. Because it is not possible to do genetic screens for long-lived humans, we are doing genetic screens in human cells instead. Our experimental strategy is based on the observation that all FOXO- dependent life-extending pathways tested so far (as well as many other life-extension pathways) increase resistance to oxidative stress. Although the role of oxidative stress resistance in life extension is not clear, the correlation is tight enough that in many model organisms, screens for oxidative stress resistance have yielded long-lived mutants. Therefore, to obtain a set of potential human longevity genes, the Kenyon lab has carried out a genome-wide siRNA screen for oxidative stress resistance in a human primary cell line. The gene hits include known C. elegans FOXO regulators, regulators of other longevity proteins such as TOR and NRF2, and new genes as well. From this set, the Kenyon lab will identify good candidates for new human longevity and healthspan genes. To do this, they will determine which knockdowns trigger other correlates of longevity, such as xenobiotic resistance or autophagy. In addition, they will ask which knockdowns perturb the activities of FOXO3A, TOR or NRF2. Finally, to link these genes to longevity, they will test for their ability to influence lifespan in C. elegans and for their altered expression in centenarian families. This fresh approach will define new potential drug targets for extending the youthful and productive years of human life, and for delaying age-related diseases such as cancer, heart disease and/or neurodegenerative disease.