Regulation of the transcription factor HLH-30/TFEB in aging

项目总结/摘要 自噬是一种细胞再循环过程，在衰老和年龄相关疾病中起着关键作用。自噬 最初被描述为主要在翻译后水平调节的过程，但在转录水平上起作用。 最近，监管开始出现。相对较新的概念，转录调控的， 自噬过程提供了一个重要的新切入点， 疾病 最近在哺乳动物中的研究已经发现螺旋-环-螺旋转录因子EB（TFEB）是一种中枢转录因子， 自噬的调节器。我们报道了C.线虫HLH-30蛋白是TFEB的功能性直向同源物 并在C.线虫寿命测定（Lapierre等，Nature Communications，2013）。 具体地说，我们已经发现，HLH-30是所需的寿命延长观察到的六个独立的 长寿模型，所有这些都是已知的依赖于自噬。值得注意的是，HLH-30定位于 这些长寿突变体的肠细胞核。重要的是，我们发现TFEB与 在哺乳动物寿命模型中调节，表明TFEB的寿命功能也是保守的。 因此，我们假设TFEB是调节衰老的核心参与者，至少部分是通过协调 自噬通量，因此，代表了一个有吸引力的长寿因素，以分子理解， 设计促进健康和防止衰老的方法。 为了解决这一假设，我们建议使用尖端的遗传学和生物化学相结合的方法， 方法来了解空间和核的要求HLH-30功能在C。线虫寿命 (Aim 1）;以解决下游靶基因，除了那些被确定为在自噬中起作用的基因，是否对 长寿（目标2）;最后，确定自噬和HLH的遗传和药理学调节因子。 30/TFEB功能（目标3）。 这些研究是重要的，因为它们是第一个确定新的遗传调节因子和翻译后调节因子。 HLH-30/TFEB的改进，它们将为未来的研究提供重要信息。 分析哺乳动物TFEB，自噬以及代谢的中央调节器，在有机体老化。 此外，我们的研究是创新的，因为我们采用C。线虫和哺乳动物细胞， 快速识别在衰老中起作用的新的保守基因，并开发药理学解决方案， 增强自噬，以预防与年龄有关的疾病。

PROJECT SUMMARY/ABSTRACT Autophagy is a cellular recycling process with critical roles in aging and age-related diseases. Autophagy was originally described as a process mainly regulated at the post-translational level, but a role for transcriptional regulation has recently started to emerge. The relatively new concept of transcriptional regulation of the autophagy process provides an important new entry point toward developing therapies against age-related diseases. Recent studies in mammals have uncovered that the helix-loop-helix transcription factor EB (TFEB) is a central regulator of autophagy. We have reported that the C. elegans HLH-30 protein is a functional ortholog of TFEB and plays a broad role in C. elegans lifespan determination (Lapierre et al., Nature Communications, 2013). Specifically, we have found that HLH-30 is required for the lifespan extension observed in six independent longevity models, all of which are also known to be dependent on autophagy. Notably, HLH-30 is localized in the nucleus of intestinal cells in all of these long-lived mutants. Importantly, we have found TFEB to be similarly regulated in a mammalian longevity model suggesting that the longevity function of TFEB is also conserved. Therefore, we hypothesize that TFEB is a central player that modulates aging, at least in part by coordinating autophagic flux and, as such, represents an attractive longevity factor to molecularly understand in order to design approaches to promote healthspan and prevent aging. To address this hypothesis, we propose to use a combination of cutting-edge genetics and biochemical approaches to understand the spatial- and nuclear requirements for HLH-30 function in C. elegans lifespan (Aim 1); to address if downstream target genes besides those identified to function in autophagy are critical for longevity (Aim 2); and, finally, to identify genetic and pharmacological regulators of autophagy and HLH- 30/TFEB function (Aim 3). These studies are significant as they are the first to identify novel genetic regulators and post-translational modifications of HLH-30/TFEB in an unbiased fashion, and they will generate important information for future analysis of mammalian TFEB, a central regulator of autophagy as well as metabolism, in organismal aging. Furthermore, our studies are innovative because we employ C. elegans and mammalian cells in parallel to rapidly identify novel, conserved genes with roles in aging and to develop pharmacological solutions to enhance autophagy in order to prevent age-related diseases.