Mechanisms Specific to the Beneficial Effects of Dietary Restriction

项目摘要 饮食限制（DR）是促进健康衰老和抵抗年龄发病的最有效方法 在动物模型中。然而，DR的治疗潜力受到相关负面生理效应的限制， 包括生长、免疫力和生殖能力受损。尽管DR的营养感应介质 已经鉴定的，如mTOR，FOXO/As和sirtuins，这些中心节点概括了整个 反应，使它们成为次优治疗靶点。我们的长期目标是发现 专门介导DR的促长寿效应的机制， 治疗学DR的关键介质是AMP活化蛋白激酶（AMPK），其是一种细胞燃料计量器， 当能量水平低的时候。然而，与DR一样，AMPK以受损的生长为代价延长寿命， 生殖 本申请的目的是使用遗传上易处理的模型系统C。elegans识别 AMPK和DR特异性介导长寿的机制，以阐明第一个分子机制， 目标，概括只有亲健康的影响，DR的中心假设是，有益的， DR的有害影响可以被分开。为了支持这一假设，在某些特定的氨基酸组合， 最近的研究表明，这种饮食可以延长寿命，同时保持正常的繁殖， DR对寿命的积极影响不需要强制性的有害副作用。此外，在此前的 我们证明，转录辅激活因子和AMPK直接靶向CRTC-1， 将AMPK的长寿效应与其他负面副作用如发育迟缓分开， 生殖抑制和减小的身体尺寸。 我们现在试图确定CRTC-1特异性介导长寿的机制。这样做的理由 在我们从DR中产生可行的治疗方法用于临床应用之前，我们必须首先确定 这些机制1）仅概括了DR的积极作用，2）在生命后期应用时有效， 年龄相关疾病的诊断。基于强有力的初步数据，我们将测试三个具体目标。第一章 我们将研究神经元CRTC-1在促进健康衰老中的功能作用。2)我们将使用CRISPR 编辑内源性AMPK靶点，以描述该关键能量下游的其他途径的作用 传感器3）我们将确定线粒体和过氧化物酶体重构在AMPK和DR寿命中的贡献。 总的来说，我们希望这项工作能够提供第一个解偶联阳性的分子途径的例子。 以及DR的负面影响，这是将DR研究转变为促进健康人类衰老的关键一步。

PROJECT SUMMARY Dietary restriction (DR) is the most potent method for promoting healthy aging and age-onset disease resistance in animal models. However, DR’s therapeutic potential is limited by associated negative physiological effects, including impaired growth, immunity and reproductive capacity. Although nutrientsensing mediators of DR have been identified, such as mTOR, FOXO/As and the sirtuins, these central nodes recapitulate the entirety of the response, making them sub-optimal therapeutic targets. Our long-term objective is to uncover molecular mechanisms that specifically mediate only the pro-longevity effects of DR to develop optimal therapeutics. A key mediator of DR is AMP-activated protein kinase (AMPK), a cellular fuel gauge activated when energy levels are low. However, like DR, AMPK increases lifespan at the cost of impaired growth and reproduction. The objective in this application is to use the genetically tractable model system C. elegans to identify mechanisms by which AMPK and DR specifically mediates longevity, in order to elucidate the first molecular targets that recapitulate only the pro-health effects of DR. The central hypothesis is that beneficial and detrimental effects of DR can be uncoupled. In support of this hypothesis, specific amino acid combinations in the diet have recently been shown to increase lifespan while maintaining normal reproduction, establishing that the positive effects of DR on lifespan do not require obligate detrimental side effects. In addition, in the previous funding period we demonstrated that the transcriptional coactivator and AMPK direct target CRTC-1 could uncouple the longevity effects of AMPK from other negative side effects such as developmental delay, reproduction suppression and reduced body size. We now seek to identify the mechanisms by which CRTC-1 specifically mediates longevity. The rationale for this project is that, before we can generate viable therapies from DR for clinical application we must first identify mechanisms that 1) recapitulate only the positive effects of DR and 2) are effective when applied late in life, post-diagnosis of age-related disease. Based upon strong preliminary data we will test three specific aims. 1) We will examine the functional role of neuronal CRTC-1 in promoting healthy aging. 2) We will use CRISPR editing of endogenous AMPK targets to delineate the roles of additional pathways downstream of this key energy sensor 3) We will define the contribution of mitochondrial and peroxisome remodeling in AMPK and DR longevity. Collectively, we expect this work to provide the first example of molecular pathways that uncouple the positive and negative effects of DR, a critical step in transitioning DR research to the promotion of healthy human aging.