Role of LIPL-4 in lysosomal lipolysis and aging

LIPL-4 在溶酶体脂肪分解和衰老中的作用

• 批准号: 9217535
• 负责人: Louis Rene Lapierre
• 金额: $ 23.7万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9217535
• 关键词: Acid Lipase; Address; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Animals; Autophagocytosis; Autophagosome; Biological Assay; Caenorhabditis elegans; Cell Culture Techniques; Cells; Chemicals; Cholesterol; Cholesterol Esters; Coupled; Data; Development; Disease; Disease model; Enzymes; Functional disorder; Genes; Genetic Screening; Grant; Homeostasis; Human; Hydrolysis; Impairment; Insulin; Lead; Learning; Link; Lipase; Lipids; Lipolysis; Longevity; Lysosomes; Mammalian Cell; Mediating; Metabolism; Methods; Microscopy; Modeling; Molecular; Mus; Nematoda; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nutrient; Organelles; Organism; Paralysed; Pathologic; Pathway interactions; Pharmacology; Phase; Physiological; Process; Proteins; Proteomics; Publishing; Recycling; Reporter; Reporting; Research; Research Proposals; Risk; Role; Scientist; Senile Plaques; Signal Pathway; Signal Transduction; Sirolimus; Site; System; Testing; Therapeutic; Translating; Vesicle; Work; Yeasts; abeta toxicity; age related; aging population; base; cholesterol trafficking; experimental study; fly; high throughput screening; innovation; insulin signaling; lipid metabolism; new technology; novel; novel therapeutics; overexpression; prevent; proteostasis; sensor; skills; sterol esterase; therapeutic target; transcription factor

PROJECT SUMMARY Dysfunctions in the autophagy/lysosomal pathway are pathologically significant in the development of age-related diseases, such as neurodegeneration. In C. elegans, several longevity models rely on increased autophagy for lifespan extension, suggesting a critical role for autophagy in aging. Animals can also enjoy longer lifespan when the nutrient-sensor TOR, a negative regulator of autophagy, is inhibited. Nonetheless, how autophagy mediates its beneficial effects is poorly understood. We recently reported that autophagy could be induced by over-expressing the putative lysosomal lipase LIPL-4, which resulted in a significant lifespan extension, enhanced lipolysis and altered TOR signaling, suggesting a link between lipid metabolism, autophagy and aging. LIPL-4 displays strong homology with human lysosomal acid lipase (LAL), a key enzyme in the hydrolysis of cholesterol via autophagy. Notably, impaired LAL-mediated cholesterol processing has been linked to the development of Alzheimer's disease. My new results show that over-expressing LIPL-4 ameliorates Aβ toxicity in a C. elegans model of Alzheimer's disease. Therefore, this proposal will test the hypothesis that LIPL-4, similar to LAL, mediates lysosomal lipid hydrolysis and will aim to elucidate how LIPL-4 modulates autophagy and mitigates Aβ toxicity. In Aim 1, I will confirm the intracellular site of action of LIPL-4 and determine its relationship to TOR signaling. In Aim 2, I will test whether LIPL-4 and LAL are functionally interchangeable in C. elegans. The mechanism of action by which LIPL-4 induces autophagy and modulates aging will also be elucidated. In Aim 3, I will investigate how LIPL-4 mediates a delay in the onset of Alzheimer's disease in C. elegans. I will also perform a high-throughput screen (HTS) to discover novel and specific candidate that activates LAL-mediated lipolysis, as a strategy against neurodegeneration. By determining the role of lysosomal lipolysis in aging, my proposal will provide a basis on which novel drugs can be discovered to prevent Alzheimer's disease. The 2-year postdoctoral K99 phase will consist in the characterization of the role of LIPL-4 in lysosomal function, lipid metabolism and aging. Cell-based assay reporter systems compatible with HTS will be used to find novel drugs to enhance LAL expression. The 3-year independent R00 phase will serve to further understand the role of LIPL-4 in lysosomal lipolysis, lipid signaling and aging and expand into studies on lipid dynamics, metabolism and proteostasis. Lead candidate activators of LAL will be validated using Alzheimer's disease model in C. elegans and cell culture models. This proposal includes cutting-edge approaches, such as proteomic analyses, CARS microscopy and HTS combined with the innovative use of disease models in C. elegans. In summary, the K99/R00 grant represents a unique opportunity for me to learn new technologies and develop my professional skills to successfully transition into an independent scientist in aging research.

项目摘要 自噬/溶酶体途径的功能障碍在发育中具有病理意义 与年龄有关的疾病，例如神经变性。在秀丽隐杆线虫中，几种寿命模型依靠 自噬增加了寿命延长，这表明自噬在衰老中起着至关重要的作用。动物 当营养传播器TOR（一种自动噬菌体的负调节器）为，也可以享受更长的寿命 抑制。尽管如此，自动噬如何培养其有益效果。我们最近 据报道，可以通过过度表达假定的溶酶体脂肪酶Lipl-4，可以诱导自噬。 这导致了显着的寿命延长，增强的脂解和TOR信号的改变， 提出脂质代谢，自噬和衰老之间的联系。 LIPL-4显示强大的同源性 与人类溶酶体脂肪酶（LAL）一起，这是一种通过自噬水解胆固醇的关键酶。 值得注意的是，LAL介导的胆固醇加工受损与发展有关 阿尔茨海默氏病。我的新结果表明，过度表达Lipl-4可以缓解C中的Aβ毒性。 阿尔茨海默氏病的秀丽隐杆线模型。因此，该提案将检验以下假设：lipl-4， 与LAL相似，介导溶酶体脂质水解，并将旨在阐明LIPL-4如何调节 自噬和减轻Aβ毒性。在AIM 1中，我将确认Lipl-4和 确定其与TOR信号的关系。在AIM 2中，我将测试Lipl-4和LAL是否在功能上是 可在秀丽隐杆线虫中互换。 Lipl-4诱导自噬和的作用机制 调节衰老也将被阐明。在AIM 3中，我将研究Lipl-4如何介导 秀丽隐杆线虫中阿尔茨海默氏病的发作。我还将执行高通量屏幕（HTS） 发现激活LAL介导的脂解的新颖和特定候选者，作为反对的策略 神经变性。通过确定溶酶体脂解在衰老中的作用，我的建议将提供 可以发现新型药物以防止阿尔茨海默氏病。 两年的博士后K99阶段将包括Lipl-4在 溶酶体功能，脂质代谢和衰老。基于细胞的测定报告机系统与 HTS将用于寻找新的药物以增强LAL表达。三年独立R00阶段 将进一步了解LIPL-4在溶酶体脂解，脂质信号传导和衰老中的作用 扩展到有关脂质动力学，代谢和蛋白质的研究。 LAL的主要候选激活剂 将使用秀丽隐杆线虫和细胞培养模型中的阿尔茨海默氏病模型进行验证。这个建议 包括尖端方法，例如蛋白质组学分析，汽车显微镜和HTS组合 通过在秀丽隐杆线虫中疾病模型的创新使用。总而言之，K99/R00赠款代表 我有独特的机会学习新技术并发展我的专业技能以成功 过渡到衰老研究的独立科学家。

项目成果

Visible light reduces C. elegans longevity.

• DOI: 10.1038/s41467-018-02934-5
• 发表时间: 2018-03-02
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: De Magalhaes Filho CD;Henriquez B;Seah NE;Evans RM;Lapierre LR;Dillin A
• 通讯作者: Dillin A

Emerging topics in C. elegans aging research: Transcriptional regulation, stress response and epigenetics.

• DOI: 10.1016/j.mad.2018.08.001
• 发表时间: 2019-01
• 期刊: Mechanisms of ageing and development
• 影响因子: 5.3
• 作者: Denzel MS;Lapierre LR;Mack HID
• 通讯作者: Mack HID

Give me a SINE: how Selective Inhibitors of Nuclear Export modulate autophagy and aging.

• DOI: 10.1080/23723556.2018.1502511
• 发表时间: 2018
• 期刊: Molecular & cellular oncology
• 影响因子: 2.1
• 作者: Kumar AV;Thakurta TG;Silvestrini MJ;Johnson JR;Reenan RA;Lapierre LR
• 通讯作者: Lapierre LR

Reduced ech-6 expression attenuates fat-induced lifespan shortening in C. elegans.

• DOI: 10.1038/s41598-022-07397-9
• 发表时间: 2022-03-01
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Liu YJ;Gao AW;Smith RL;Janssens GE;Panneman DM;Jongejan A;van Weeghel M;Vaz FM;Silvestrini MJ;Lapierre LR;MacInnes AW;Houtkooper RH
• 通讯作者: Houtkooper RH