Regulation Of Metabolism And Gene Expression By Iron-Sulfur Clusters - Supplement

原始获奖项目摘要(添加粗体以强调与设备请求相关的内容) 铁-硫簇(ISCs)是一种必需的蛋白质辅因子，其失调与广泛的 包括弗里德里希共济失调在内的衰弱疾病。这种病理在一定程度上是由于在铁中使用ISCs- 反应蛋白(IRPS)，在铁饥饿反应中控制mRNAs的翻译。我们最近 研究发现，通过抑制ISC的生物合成，我们可以有力地激活IRP介导的铁饥饿反应， 通过铁下垂使癌细胞对细胞死亡敏感。我们的初步数据，这构成了我们 应用程序，指出了ISCs对IRP2的一种意想不到的调节模式，并通过 哪些ISCs调节铁饥饿反应，从而影响人类疾病，目前尚不清楚。我们的 长期目标是剖析iSCs调节细胞对变化的反应的机制。 铁和氧的可利用性以及这种反应是如何由IRPS介导的。我们预计这些发现 将导致识别先前ISC参与不清楚的疾病的病理，到新的 治疗ISC调节失调的疾病，并将促进新的方法使癌细胞对 铁性下垂。这项资助的目的是剖析ISCs如何控制IRP2的激活，并全面地 定义IRP1和IRP2的目标以及它们被激活的条件。我们最重要的是 假设ISCs整合铁和氧水平的输入来影响特定的翻译反应 通过对IRPS的差异调控。我们的基本原理是确定内部审查制度的具体机制 被激活，它们激活的靶点将使发现治疗癌症和 ISC代谢紊乱。我们的具体目标将检验以下假设：(目标1)ISCs可以激活 IRP2介导的铁饥饿反应；(AIM 2)IRPS表现出不同的IRE结合 以响应铁和氧气水平的调节。在完成这些目标后，我们将(1)获得 了解细胞如何感知铁和氧水平，并使用ISC传感器整合这些输入 以及(2)鉴定IRP调控的靶基因及其调控条件。这一贡献是 重要的是因为IRPS的调节失调在人类的病理中经常发生，并诱导 铁饥饿反应使癌细胞对铁性细胞死亡敏感。这项研究具有创新性，因为我们 挑战细胞铁感应的范式，以达到细胞响应的综合机制 这种重要营养素水平的变化，因为我们利用迄今为止独特的方法来识别 受铁饥饿反应调节的RNA，并通过上游刺激定义它们的激活。结果是 这项研究的前景打破了我们对铁感应的理解，并对治疗糖尿病具有广泛的意义 癌症以及与铁代谢和储存缺陷有关的人类疾病。

Project Abstract from Original Award (bold text added for emphasis related to the equipment request) Iron-sulfur clusters (ISCs) are essential protein cofactors whose dysregulation is linked to a wide range of debilitating diseases including Friedreich’s ataxia. This pathology is due in part to the use of ISCs in iron- responsive proteins (IRPs), which control the translation of mRNAs in the iron-starvation response. We recently found that by suppressing ISC biosynthesis we can robustly activate this IRP-mediated iron-starvation response, sensitizing cancer cells to cell death by ferroptosis. Our preliminary data, which form the premise of our application, point to an unexpected mode of IRP2 regulation by ISCs and demonstrate that the mechanism by which ISCs modulate the iron-starvation response, and therefore impact human disease, remains unclear. Our long-term goal is to dissect the mechanisms by which ISCs regulates cellular responses to changing iron and oxygen availability and how this response is mediated by IRPs. We anticipate that these discoveries will lead to the identification of pathologies for diseases where prior ISC involvement was unclear, to new treatments for diseases of ISC dysregulation, and will facilitate novel methods to sensitize cancer cells to ferroptosis. The objective of this grant is to dissect how ISCs control IRP2 activation and to comprehensively define the targets of IRP1 and IRP2 and the conditions under which they are activated. Our overarching hypothesis is that ISCs integrate iron and oxygen level inputs to effect specific translational responses via differential regulation of IRPs. Our rationale is that identification of the specific mechanisms by which IRPs are activated and the targets that they activate will enable the discovery of novel strategies to treat cancer and disorders of ISC metabolism. Our specific aims will test the following hypotheses: (Aim 1) ISCs can activate the iron-starvation response through an IRP2-mediated mechanism; (Aim 2) IRPs exhibit differential IRE binding in response to iron and oxygen level modulation. Upon completion of these aims we will (1) gain an understanding of how cells sense iron and oxygen levels and integrate these inputs using ISC sensors and (2) identify IRP regulated target genes and the conditions in which they are regulated. This contribution is significant because dysregulation of IRPs occurs frequently in human pathologies, and inducing activation of the iron starvation response sensitizes cancer cells to ferroptotic cell death. This research is innovative because we challenge paradigms in cellular iron-sensing to arrive at a comprehensive mechanism by which cells respond to changes in the level of this important nutrient, and because we utilize heretofore unique approaches to identify RNAs regulated by the iron starvation response and define their activation by upstream stimuli. The outcomes of this study promise disrupt our understanding of iron sensing and have broad implications for the treatment of cancer as well as human diseases related to defects in iron metabolism and storage.