Mechanisms Specific to the Beneficial Effects of Dietary Restriction

项目总结 限制饮食(DR)是促进健康老龄化和抗老年性疾病的最有效的方法 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的营养感官介质有 例如mTOR、FOXO/AS和sirtuins，这些中央节点概括了 反应，使它们成为次优的治疗靶点。我们的长期目标是发现分子 仅专门调节DR的长寿效应的机制，以开发最佳 治疗学。DR的一个关键介质是AMP激活的蛋白激酶(AMPK)，这是一种激活的细胞燃料计 当能量水平较低时。然而，像DR一样，AMPK延长了寿命，但代价是生长受损和 繁殖。 这个应用程序的目标是使用遗传易处理的模型系统线虫来识别 AMPK和DR特异性调节长寿的机制，以阐明第一个分子 目标只概括了博士对健康的影响。中心假设是有益的和 DR的有害影响可能是分离的。为了支持这一假说，体内特定的氨基酸组合 最近，这种饮食被证明在保持正常生殖的同时延长了寿命，证实了 DR对寿命的积极影响不需要额外的有害副作用。此外，在之前的 资助期间，我们证明了转录共激活因子和AMPK直接靶标CRTC-1可以 将AMPK的长寿效应与其他负面副作用，如发育迟缓， 生殖抑制和体型缩小。 我们现在试图确定CRTC-1具体调节长寿的机制。这样做的理由是 项目是，在我们可以从DR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 1)仅概括DR的积极影响的机制和2)在生命后期应用时有效的机制， 年龄相关疾病的诊断后。基于强大的初步数据，我们将测试三个具体目标。1) 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靶标以描绘该关键能量下游的额外通路的作用 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长寿中的作用。 总而言之，我们希望这项工作将提供第一个解偶联阳离子的分子途径的例子 以及DR的负面影响，这是将DR研究过渡到促进人类健康老龄化的关键一步。

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.