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

原授标项目摘要（为强调与设备申请相关的内容而添加粗体文本） 铁硫簇（ISCs）是一种重要的蛋白质辅因子，其调节异常与多种疾病有关。 包括弗里德赖希共济失调在内的衰弱性疾病。这种病理学部分是由于在铁中使用ISCs- 反应蛋白（IRP），其控制铁饥饿反应中mRNA的翻译。我们最近 发现通过抑制ISC生物合成，我们可以强烈激活这种IRP介导的铁饥饿反应， 使癌细胞对由铁凋亡引起的细胞死亡敏感。我们的初步数据，这构成了我们的前提， 应用程序，指出一个意想不到的模式IRP2的调控，由ISCs和证明，该机制， 哪些ISCs调节铁饥饿反应并因此影响人类疾病仍不清楚。我们 长期目标是剖析ISCs调节细胞对变化的反应的机制， 铁和氧的可用性，以及这种反应是如何介导的IRP。我们预计这些发现 将导致确定疾病的病理，其中先前ISC参与不清楚，以新的 治疗ISC失调的疾病，并将促进新的方法，使癌细胞敏感， 铁性下垂这项资助的目的是剖析ISC如何控制IRP2激活，并全面 定义IRP1和IRP2的靶点以及它们被激活的条件。我们的总体 假设ISCs整合铁和氧水平输入以影响特异性翻译反应 通过IRP的差异调节。我们的基本原理是，识别IRP的具体机制， 被激活，它们激活的靶点将有助于发现治疗癌症的新策略， ISC代谢紊乱。我们的具体目标将测试以下假设：（目标1）ISCs可以激活 通过IRP 2介导的机制的铁饥饿反应;（目的2）IRP表现出差异IRE结合 以响应铁和氧水平调节。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 和（2）鉴定IRP调节的靶基因和它们被调节的条件。这种贡献 重要的是，因为IRP的失调经常发生在人类病理学中，并且诱导IRP的激活。 铁饥饿反应使癌细胞对铁凋亡细胞死亡敏感。这项研究是创新的，因为我们 挑战细胞铁传感的范式，以达到细胞响应的全面机制， 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 RNA受铁饥饿反应的调节，并通过上游刺激来定义其激活。成果 这项研究的结果有望打破我们对铁传感的理解，并对治疗 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.