Novel anti-NPC aggregation strategy against brain ischemia-reperfusion injury

中风是一种毁灭性的疾病，影响着数百万美国人。但大多数 先前的中风药物开发计划由于各种原因而不成功。它们中的一个可以是 合适的治疗靶点仍有待确定。我们最近的研究表明， 新生肽链（NPCs）的聚集可能在缺血-再灌注损伤中起作用。NPC，即， 新合成或部分合成的多肽是未折叠蛋白质的主要来源， 正常细胞中的毒性聚集。为了避免毒性聚集，细胞NPC必须受到保护， 分子伴侣我们最新的研究表明，脑缺血损伤多组分子， 伴侣蛋白，导致大规模的NPC聚集与ER，高尔基体和线粒体结构， 再灌注NPC聚集体相关细胞器的数量逐渐增加， 脑缺血后发生神经元死亡。尽管我们的研究强烈支持这一假设，但 仍然是一个很大程度上悬而未决的问题，因为没有研究表明，阻止NPC聚集 保护脑免受缺血-再灌注损伤，以及用于控制NPC聚集的分子靶标 仍有待确认。此外，研究这些基本的翻译能力可能是至关重要的 科学发现。我们最近发现，新开发的eIF 4 E抑制剂具有很强的抗- NPC聚集效应，并在动物缺血模型中提供强大的神经保护作用。Aim 1设计 对于新假设的概念验证研究，即大规模NPC聚集在 eIF 4 E是一个新的、最好的抗NPC聚集的治疗靶点。这 一个新的假说以前没有被验证过，但对于开发新的治疗策略至关重要 抗缺血再灌注损伤。在这个目标中，我们将使用新开发的引发抑制剂， 几项新技术：（i）确定针对大规模NPC聚集的最佳治疗靶点; 以及（ii）研究新的抗NPC聚集策略是否保护蛋白质质量控制 系统和防止脑缺血后多个细胞器衰竭。目的2提出临床研究 新的抗NPC聚集策略的翻译能力。这一目标基于新的确凿证据， 缺血后和腹膜内注射eIF 4 E抑制剂在动物脑中提供稳健的神经保护 缺血模型。我们将初步确定最佳治疗靶点，然后进行全面的 根据STAIR标准和新的NIH指南Rigor和 复制品包括：（i）两种临床相关的局灶性缺血模型;（ii）雄性和雌性，（iii）3- 和12个月年龄组，（iv）缺血后治疗，（v）7天和28天终点，以及（vi）进行 彻底的分子病理学和神经学评估这些研究将捕获许多临床 机制和神经保护的相关方面。

Project Summary: Stroke is a devastating disease affecting millions of Americans. However, most prior stroke drug development programs were unsuccessful for a variety of reasons. One of them may be that the appropriate therapeutic targets remain to be identified. Our recent studies show that massive aggregation of nascent peptide chains (NPCs) may play a role in ischemia-reperfusion injury. NPCs, i.e., newly or partially synthesized polypeptides, are the major source of unfolded proteins and highly prone to toxic aggregation in a normal cell. To avoid toxic aggregation, cellular NPCs must be protected by molecular chaperones. Our latest studies show that brain ischemia damages multiple groups of molecular chaperones, resulting in massive NPC aggregation with the ER, Golgi, and mitochondria structures during reperfusion. The amounts of NPC aggregate-associated organelles increase progressively until delayed neuronal death occurs after brain ischemia. Although our studies strongly support this hypothesis, this remains a largely unsettled issue because no study has ever shown that blocking of NPC aggregation protects the brain from ischemia-reperfusion injury, and molecular target(s) for managing NPC aggregation remain to be identified. Furthermore, it may be vitally important to study the translation ability of these basic science discoveries. We have recently found that the newly developed eIF4E inhibitors have strong anti- NPC aggregation effects and offer robust neuroprotection in animal ischemia models. Aim 1 is designed for proof-of-concept studies of the novel hypothesis that massive NPC aggregation plays a key role in ischemia-reperfusion injury, and eIF4E is a new and best therapeutic target against NPC aggregation. This new hypothesis has not been tested previously but is essential to develop new therapeutic strategies against ischemia-reperfusion injury. In this Aim, we will use newly developed initiation inhibitors and several new technologies: (i) to identify the best therapeutic target(s) against massive NPC aggregation; and (ii) to study whether the novel anti-NPC aggregation strategy protects the protein quality control systems and prevents multiple organelle failure after brain ischemia. Aim 2 proposes studies of the clinical translation ability of the new anti-NPC aggregation strategy. This Aim is based on solid new evidence that postischemic and intraperitoneal injection of eIF4E inhibitors offers robust neuroprotection in animal brain ischemia models. We will initially identify the best therapeutic target(s), and then carry out comprehensive studies of the neuroprotection following the STAIR criteria and the new NIH guideline of Rigor and Reproducibility including: (i) two clinically relevant focal ischemia models; (ii) both male and female, (iii) 3- and 12-month age groups, (iv) post-ischemic treatment, (v) 7- and 28-day endpoints, and (vi) performing thorough molecular, pathological and neurological evaluations. These studies will capture many clinically relevant aspects of the mechanisms and neuroprotection.