Ferroptosis and Polyunsaturated Fatty Acid Metabolism

项目概述：我们研究计划的总体目标是开发化学工具和体内管道 阐明内源性氧化多不饱和脂肪酸（PUFAs）调节铁凋亡的机制。 铁凋亡是最近发现的一种铁依赖性非凋亡性程序性细胞死亡， 通过增加膜脂质过氧化作用。像其他程序性细胞死亡一样，铁凋亡具有广泛的影响， 对细胞和胚胎发育、细胞衰老、组织稳态和许多疾病的影响。若干关键蛋白 和调节铁凋亡的化合物，但铁凋亡的潜在机制和 其内源性介质知之甚少。这个建议是基于我们围绕PUFA的研究 氧化代谢与健康我们的研究重点是产生新的化学工具，以调查 PUFA代谢产物对人体健康的作用机制。具体来说，我们开发了多种合成方法， 以制备细胞色素P450（CYP450）PUFA代谢物。我们还设计并合成了相应的 PUFA小说模仿。我们应用这些合成的代谢物和模拟物来鉴定许多新的生物活性物质， PUFA代谢产物的影响。探讨多不饱和脂肪酸生物学效应背后的机制 代谢产物，我们建立了C.作为一种新的生物模型在我们的实验室由于巨大的遗传工具 适用于高通量筛选。所有这些特征都极大地促进了机械化 问题研究利用C.我们发现，内源性PUFAs和相应的代谢产物诱导 仅在生殖细胞和神经元中存在铁凋亡。基于这些发现，我们对此的中心假设 推测PUFA代谢物是铁凋亡的内源性脂质介质。大大推进 为了理解这种新的程序性细胞死亡，R35将解决两个问题：1） 铁凋亡的内源性信号分子和2）铁凋亡在细胞类型特异性 举止？在接下来的五年里，我们将使用PUFA代谢物，代谢物模拟物和代谢抑制剂， 我们将结合一种新的模式生物（C.以及它们庞大的遗传工具来回答 我们的科学问题，1）确定内源性多不饱和脂肪酸代谢物，触发或抑制铁凋亡和2） 开发一种新的体内平台来描绘细胞特异性铁凋亡调控途径。 了解氧化的PUFA代谢物如何在特定细胞类型中触发铁凋亡将对以下方面产生影响： 铁性下垂症如何影响人类健康该提案的成果将为我们的 未来多层次的研究。确定的脂质介质和蛋白质靶点将在 用于研究翻译和机制研究的哺乳动物模型。脂质介质和蛋白质的发现 靶点也使我们能够开发候选药物来治疗与铁凋亡相关的疾病。发达 转基因菌株和体内平台将使我们能够鉴定新的信号分子或蛋白质靶点， 调节细胞类型特异性铁凋亡。我们将继续开发新的化学工具来研究铁凋亡。

Project Summary: The overall goal of our research program is to develop chemical tools and an in vivo pipeline to delineate mechanisms of ferroptosis regulation by endogenous oxidized polyunsaturated fatty acids (PUFAs). Ferroptosis is a recently discovered iron-dependent non-apoptotic programmed cell death that is characterized by an increase in membrane lipid peroxidation. Like other programmed cell death, ferroptosis has a broad impact on cell and embryo development, cell senescence, tissue homeostasis, and many diseases. Several key proteins and compounds that regulate ferroptosis have been identified, but the underlying mechanism of ferroptosis and its endogenous mediators are poorly understood. This proposal is based on our research centered around PUFA oxidative metabolism and health. Our research focuses on generating novel chemical tools to investigate the mechanism of PUFA metabolites on human health. Specifically, we have developed multiple synthetic methods to prepare cytochrome P450 (CYP) PUFA metabolites. We also designed and synthesized the corresponding PUFA novel mimics. We applied these synthesized metabolites and mimics to identify many new biological effects of PUFA CYP metabolites. To investigate the mechanism behind the biological effects of CYP PUFA metabolites, we established C. elegans as a novel biological model in our lab due to the vast genetic tools available and the adaptability for high-throughput screening. All of these features greatly facilitate mechanistic studies. Using C. elegans, we found that endogenous PUFAs and the corresponding CYP metabolites induce ferroptosis only in the germline and neurons. Based on these findings, our central hypothesis for this proposal is that PUFA metabolites are endogenous lipid mediators of ferroptosis. To significantly advance the understanding of this novel programmed cell death, this R35 will address two questions: 1) What are the endogenous signaling molecules of ferroptosis? And 2) How is ferroptosis regulated in a cell type-specific manner? In the next five years, we will use PUFA metabolites, metabolite mimics, and metabolic inhibitors that we will synthesize in combination with a novel model organism (C. elegans) and their vast genetic tools to answer our scientific questions by 1) identifying endogenous PUFA metabolites that trigger or inhibit ferroptosis and 2) developing a novel in vivo platform to delineate cell-specific ferroptosis regulatory pathways. Understanding how oxidized PUFA metabolites trigger ferroptosis in specific cell types will have implications for how ferroptosis may affect human health. The outcomes from this proposal will build a strong foundation for our future research on multiple levels. The identified lipid mediators and protein targets will be further tested in mammalian models for research translation and mechanistic studies. The discovery of lipid mediators and protein targets also allows us to develop drug candidates to treat diseases related to ferroptosis. The developed transgenic strains and in vivo platform will allow us to identify novel signaling molecules or protein targets that regulate cell-type specific ferroptosis. We will continue to develop novel chemical tools to investigate ferroptosis.