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.

项目概要 自噬/溶酶体途径的功能障碍在发育中具有病理意义 与年龄相关的疾病，例如神经退行性疾病。在秀丽隐杆线虫中，几种长寿模型依赖于 增加自噬以延长寿命，表明自噬在衰老中发挥着关键作用。动物 当营养传感器 TOR（自噬的负调节因子）处于正常状态时，细胞也可以享有更长的寿命。 抑制。尽管如此，人们对自噬如何介导其有益作用仍知之甚少。我们最近 报道称，自噬可以通过过度表达假定的溶酶体脂肪酶 LIPL-4 来诱导， 这导致寿命显着延长、脂肪分解增强并改变 TOR 信号传导， 表明脂质代谢、自噬和衰老之间存在联系。 LIPL-4表现出很强的同源性 含有人溶酶体酸性脂肪酶 (LAL)，这是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以下疾病的发生有关： 阿尔茨海默病。我的新结果表明，过度表达 LIPL-4 可改善 C. 阿尔茨海默病的线虫模型。因此，本提案将检验 LIPL-4 的假设， 与 LAL 类似，介导溶酶体脂质水解，旨在阐明 LIPL-4 如何调节 自噬并减轻 Aβ 毒性。在目标 1 中，我将确认 LIPL-4 的细胞内作用位点和 确定其与 TOR 信号传导的关系。在目标 2 中，我将测试 LIPL-4 和 LAL 是否功能正常 在秀丽隐杆线虫中可以互换。 LIPL-4诱导自噬和自噬的作用机制 调节衰老也将得到阐明。在目标 3 中，我将研究 LIPL-4 如何调节延迟 秀丽隐杆线虫中阿尔茨海默病的发病。我还将执行高通量筛选 (HTS) 发现激活 LAL 介导的脂肪分解的新颖且特定的候选物，作为对抗的策略 神经变性。通过确定溶酶体脂肪分解在衰老中的作用，我的建议将提供 在此基础上发现预防阿尔茨海默病的新药。 为期 2 年的博士后 K99 阶段将包括描述 LIPL-4 在 溶酶体功能、脂质代谢和衰老。基于细胞的检测报告系统兼容 HTS 将用于寻找增强 LAL 表达的新药物。 3年独立R00阶段 将有助于进一步了解 LIPL-4 在溶酶体脂肪分解、脂质信号传导和衰老中的作用 扩展到脂质动力学、代谢和蛋白质稳态的研究。 LAL 的主要候选激活剂 将使用线虫中的阿尔茨海默病模型和细胞培养模型进行验证。这个提议 包括尖端方法，例如蛋白质组分析、CARS 显微镜和 HTS 相结合 在线虫疾病模型的创新应用。总之，K99/R00 补助金代表 为我提供学习新技术和发展专业技能以取得成功的独特机会 转变为衰老研究领域的独立科学家。

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

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

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