Collins Colm B, Strassheim Derek, Aherne Carol M, Yeckes Alyson R, Jedlicka Paul, de Zoeten Edwin F
*Mucosal Inflammation Program, Department of Pediatrics, University of Colorado School of Medicine, Children's Hospital Colorado, Digestive Health Institute, Aurora, Colorado; †Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado; and ‡Department of Pathology, University of Colorado School of Medicine, Aurora, Colorado.
Inflamm Bowel Dis. 2014 Apr;20(4):685-94. doi: 10.1097/01.MIB.0000442839.28664.75.
Inflammatory bowel diseases are chronic intestinal inflammatory diseases thought to reflect a dysregulated immune response. Although antibody-based inhibition of tumor necrosis factor-α (TNF-α) has provided relief to many inflammatory bowel diseases patients, these therapies are either ineffective in a patient subset or lose their efficacy over time, leaving an unmet need for alternatives. Given the critical role of the heat shock response in regulating inflammation, this study proposed to define the impact of selective inhibition of heat shock protein 90 (HSP90) on intestinal inflammation. Using multiple preclinical mouse models of inflammatory bowel diseases, we demonstrate a potent anti-inflammatory effect of selective inhibition of the HSP90 C-terminal ATPase using the compound novobiocin. Novobiocin-attenuated dextran sulfate sodium-induced colitis and CD45RB adoptive-transfer colitis through the suppression of inflammatory cytokine secretion, including TNF-α. In vitro assays demonstrate that CD4 T cells treated with novobiocin produced significantly less TNF-α measured by intracellular cytokine staining and by enzyme-linked immunosorbent assay. This corresponded to significantly decreased nuclear p65 translocation by Western blot and a decrease in nuclear factor-κB luciferase activity in Jurkat T cells. Finally, to verify the anti-TNF action of novobiocin, 20-week-old TNFΔ mice were treated for 2 weeks with subcutaneous administration of novobiocin. This model has high levels of circulating TNF-α and exhibits spontaneous transmural segmental ileitis. Novobiocin treatment significantly reduced inflammatory cell infiltrate in the ileal lamina propria. HSP90 inhibition with novobiocin offers a novel method of inflammatory cytokine suppression without potential for the development of tolerance that limits current antibody-based methods.
炎症性肠病是一类慢性肠道炎症性疾病,被认为反映了免疫反应失调。尽管基于抗体的肿瘤坏死因子-α(TNF-α)抑制疗法已使许多炎症性肠病患者病情得到缓解,但这些疗法在部分患者中无效,或随着时间推移失去疗效,因此仍存在对替代疗法的未满足需求。鉴于热休克反应在调节炎症中的关键作用,本研究旨在确定选择性抑制热休克蛋白90(HSP90)对肠道炎症的影响。我们使用多种炎症性肠病的临床前小鼠模型,证明了使用化合物新生霉素选择性抑制HSP90 C末端ATP酶具有强大的抗炎作用。新生霉素通过抑制包括TNF-α在内的炎性细胞因子分泌,减轻了葡聚糖硫酸钠诱导的结肠炎和CD45RB过继转移结肠炎。体外试验表明,用新生霉素处理的CD4 T细胞通过细胞内细胞因子染色和酶联免疫吸附测定法检测到产生的TNF-α显著减少。这与蛋白质印迹法检测到的核p65易位显著减少以及Jurkat T细胞中核因子-κB荧光素酶活性降低相对应。最后,为了验证新生霉素的抗TNF作用,对20周龄的TNFΔ小鼠皮下注射新生霉素进行为期2周的治疗。该模型具有高水平的循环TNF-α,并表现出自发性透壁节段性回肠炎。新生霉素治疗显著减少了回肠固有层的炎性细胞浸润。用新生霉素抑制HSP90提供了一种新的抑制炎性细胞因子的方法,且不存在限制当前基于抗体方法的耐受性发展的可能性。