Identification of Celastramycin as a Novel Therapeutic Agent for Pulmonary Arterial Hypertension High-Throughput Screening of 5562 Compounds
Identification of Celastramycin as a Novel Therapeutic Agent for Pulmonary Arterial Hypertension High-Throughput Screening of 5562 Compounds
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DOI：
10.1161/circresaha.119.315229
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发表时间：
2019-07-19
影响因子：
20.1
通讯作者：
Shimokawa, Hiroaki
中科院分区：
文献类型：
--
作者：
Kurosawa, Ryo;Satoh, Kimio;Shimokawa, Hiroaki
理由：肺动脉高压（PAH）的特点是肺动脉平滑肌细胞（PASMC）增殖增强，伴随炎症因子产生增加以及线粒体代谢适应过度增殖状态。然而，临床上使用的药物均以肺血管扩张为目标，对于晚期PAH患者可能效果不佳。目的：我们的目的是发现一种抑制 PASMC 增殖的治疗 PAH 的新药物。方法和结果：我们利用高通量筛选系统从原始库中筛选了5562个化合物，发现了抑制PAH患者PASMCs（PAH-PASMCs）增殖的化合物。我们发现，西拉霉素（一种最初在细菌提取物中发现的苯甲酰吡咯型化合物）以剂量依赖性方式抑制 PAH-PASMC 的增殖，对健康供体的 PASMC 的影响相对较小。然后，我们制备了25种西拉霉素类似物，并选择了先导化合物，其显着抑制PAH-PASMCs的细胞增殖并降低细胞质活性氧水平。机制分析表明，西拉霉素降低了 PAH-PASMC 中 HIF-1 α（缺氧诱导因子 1 α）和 kappa B（核因子-kappa B）的蛋白水平，前者会损害有氧代谢，后者会诱导促炎信号，从而导致炎性细胞因子的分泌减少。重要的是，西拉霉素治疗降低了 PAH-PASMC 中的活性氧水平，同时增加了 Nrf2（核因子红细胞 2 相关因子 2）的蛋白质水平，Nrf2 是细胞对抗氧化应激反应的主要调节因子。此外，西拉霉素治疗改善了线粒体能量代谢，恢复了 PAH-PASMC 中的线粒体网络形成。此外，这些西拉霉素介导的作用受到西拉霉素结合伴侣 ZFC3H1（含锌指 C3H1 结构域的蛋白）的调节。最后，西拉霉素治疗改善了 3 个实验动物模型的肺动脉高压，同时减少了肺部炎症变化。结论：这些结果表明，西拉霉素可改善肺动脉高压，减少 PAH-PASMC 的过度增殖，减少炎症和活性氧水平，并恢复线粒体能量代谢。因此，西拉霉素是一种针对 PAH 的新型药物，其目标是对 PAH-PASMC 具有抗增殖作用。
Rationale: Pulmonary arterial hypertension (PAH) is characterized by enhanced proliferation of pulmonary artery smooth muscle cells (PASMCs) accompanying increased production of inflammatory factors and adaptation of the mitochondrial metabolism to a hyperproliferative state. However, all the drugs in clinical use target pulmonary vascular dilatation, which may not be effective for patients with advanced PAH. Objective: We aimed to discover a novel drug for PAH that inhibits PASMC proliferation. Methods and Results: We screened 5562 compounds from original library using high-throughput screening system to discover compounds which inhibit proliferation of PASMCs from patients with PAH (PAH-PASMCs). We found that celastramycin, a benzoyl pyrrole-type compound originally found in a bacteria extract, inhibited the proliferation of PAH-PASMCs in a dose-dependent manner with relatively small effects on PASMCs from healthy donors. Then, we made 25 analogs of celastramycin and selected the lead compound, which significantly inhibited cell proliferation of PAH-PASMCs and reduced cytosolic reactive oxygen species levels. Mechanistic analysis demonstrated that celastramycin reduced the protein levels of HIF-1 alpha (hypoxia-inducible factor 1 alpha), which impairs aerobic metabolism, and kappa B (nuclear factor-kappa B), which induces proinflammatory signals, in PAH-PASMCs, leading to reduced secretion of inflammatory cytokine. Importantly, celastramycin treatment reduced reactive oxygen species levels in PAH-PASMCs with increased protein levels of Nrf2 (nuclear factor erythroid 2-related factor 2), a master regulator of cellular response against oxidative stress. Furthermore, celastramycin treatment improved mitochondrial energy metabolism with recovered mitochondrial network formation in PAH-PASMCs. Moreover, these celastramycin-mediated effects were regulated by ZFC3H1 (zinc finger C3H1 domain-containing protein), a binding partner of celastramycin. Finally, celastramycin treatment ameliorated pulmonary hypertension in 3 experimental animal models, accompanied by reduced inflammatory changes in the lungs. Conclusions: These results indicate that celastramycin ameliorates pulmonary hypertension, reducing excessive proliferation of PAH-PASMCs with less inflammation and reactive oxygen species levels, and recovered mitochondrial energy metabolism. Thus, celastramycin is a novel drug for PAH that targets antiproliferative effects on PAH-PASMCs.