Discovering Small Molecule Activators of Stress-responsive Signaling

维持分泌的蛋白质稳态，或蛋白质稳态，涉及平衡蛋白质的生物合成， 跨膜转移、折叠、降解等，我们假设这些对健康衰老至关重要。 由于对维持蛋白质平衡的分泌室的需求随着发育、衰老和 环境应激，哺乳动物进化出未折叠蛋白反应(UPR)应激反应信号 途径，转录调节分泌蛋白平衡网络能力，以满足需求。近期 人类遗传、化学、生物和活体证据表明，激活保护性的IRE1/XBP1s或 UPR的ATF6分支有重大的希望改善与年龄相关的分泌蛋白稳态和 纠正与多种病因疾病相关的失衡，包括系统性淀粉样变性疾病， 心血管疾病、糖尿病和神经退行性疾病，如阿尔茨海默病和 帕金森氏症。很少有化合物能够实现手臂选择性UPR激活，而那些确实会受到影响的化合物 从阻碍他们翻译发展的限制中解脱。我们利用了基于细胞的转录 记者对高通量筛选(HTS)的小型化分析，以及全细胞转录和 蛋白质组学图谱以了解转录和翻译反应的选择性 我们的点映大热。我们用药物化学阐述了有前景的化合物，建立了第一个- 具有选择性激活保护性IRE1/XBP1s或ATF6的小分子“蛋白抑制调节剂” 提高了普遍定期审议的效力和选择性的信号臂，并探讨了它们的作用机制 通过多种途径。我们将评估我们的蛋白抑制调节剂是否可以诱导保护性、ARM- 幼年和老年动物的选择性UPR激活。我们已经建立了合作来检验这一假设 我们的IRE1/XBP1和ATF6激活剂将有助于改善分泌的病理失衡 与多种疾病相关的蛋白沉积，包括系统性淀粉样变性、退行性眼球 疾病、心血管疾病和神经退行性疾病。此外，我们还将展示这些 化合物在药理上改善与阿尔茨海默病相关的两种病理表型 细胞培养模型：即Aβ和Aβ寡聚体相关的神经元细胞毒性的病理产物。我们 将向科学界提供第一批具有良好特性的小分子，这些小分子优先激活 具有确定的效力和选择性的IRE1/XBP1或ATF6 UPR转录程序。这些 化合物有可能被广泛用于治疗与年龄相关的一系列疾病 疾病。重要的是，这些化合物将向所有具有疾病模型的科学家提供 药理学上的IRE1/XBP1s或ATF6激活可能影响发病机制。是否可以使用 这些化合物提供了广泛影响科学努力的多个方面的承诺，由 包括干细胞生物学等基础科学。

The maintenance of secreted protein homeostasis, or proteostasis, involves balancing protein biosynthesis, translocation across membranes, folding, degradation, etc., which we hypothesize is critical for healthy aging. Since the demands on secretory compartments to maintain proteostasis change with development, aging, and environmental stresses, mammals evolved the Unfolded Protein Response (UPR) stress-responsive signaling pathway, which transcriptionally adjusts secretory proteostasis network capacity to meet demand. Recent human genetic, chemical biologic, and in vivo evidence shows that activating the protective IRE1/XBP1s or ATF6 arms of the UPR has significant promise to ameliorate age-related declines in secretory proteostasis and correct imbalances associated with etiologically-diverse diseases, including systemic amyloid diseases, cardiovascular disorders, diabetes, and neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Few compounds exist to achieve arm-selective UPR activation, and those that do suffer from limitations that prevent their translational development. We have leveraged cell-based transcriptional reporter assays miniaturized for high-throughput screening (HTS), along with whole cell transcriptional and proteomic profiling to understand the selectivity of the transcriptional and translational response generated by our screening hits. We have elaborated promising compounds using medicinal chemistry to establish first-in- class small molecule `proteostasis regulators' that selectively activate the protective IRE1/XBP1s or ATF6 signaling arms of the UPR with improved potency and selectivity, and we seek their mechanism of action through multiple approaches. We will assess whether our proteostasis regulators can induce protective, arm- selective UPR activation in young and old animals. We have established collaborations to test the hypothesis that our IRE1/XBP1s and ATF6 activators will be useful for ameliorating pathologic imbalances in secretory proteostasis associated with multiple diseases, including the systemic amyloidoses, degenerative eye diseases, cardiovascular disease, and neurodegenerative disorders. Furthermore, we will show that these compounds pharmacologically ameliorate two pathologic phenotypes associated with Alzheimer's disease in cell culture models: i.e., the pathologic production of Aβ and Aβ oligomer-associated neuronal cytotoxicity. We will deliver to the scientific community the first well-characterized small molecules that preferentially activate the IRE1/XBP1s or the ATF6 UPR transcriptional programs with a defined potency and selectivity. These compounds have the potential to be widely employed as therapeutics for a spectrum of age-associated diseases. Importantly, these compounds will be made available to all scientists with disease models wherein pharmacologic IRE1/XBP1s or ATF6 activation has the potential to influence pathogenesis. The availability of these compounds offers the promise to broadly influence multiple aspects of scientific endeavor funded by the NIH, including basic science such as stem cell biology.