Department of Neurosciences (Rita Levi Montalcini), University of Turin, Turin, Italy.
Department of Pharmacology, Federal University of Santa Catarina, Florianópolis, Brazil.
Front Immunol. 2022 Feb 1;13:837180. doi: 10.3389/fimmu.2022.837180. eCollection 2022.
Sepsis and septic shock are associated with high mortality and are considered one of the major public health concerns. The onset of sepsis is known as a hyper-inflammatory state that contributes to organ failure and mortality. Recent findings suggest a potential role of two non-receptor protein tyrosine kinases, namely Focal adhesion kinase (FAK) and Proline-rich tyrosine kinase 2 (Pyk2), in the inflammation associated with endometriosis, cancer, atherosclerosis and asthma. Here we investigate the role of FAK-Pyk2 in the pathogenesis of sepsis and the potential beneficial effects of the pharmacological modulation of this pathway by administering the potent reversible dual inhibitor of FAK and Pyk2, PF562271 (PF271) in a murine model of cecal ligation and puncture (CLP)-induced sepsis. Five-month-old male C57BL/6 mice underwent CLP or Sham surgery and one hour after the surgical procedure, mice were randomly assigned to receive PF271 (25 mg/kg, s.c.) or vehicle. Twenty-four hours after surgery, organs and plasma were collected for analyses. In another group of mice, survival rate was assessed every 12 h over the subsequent 5 days. Experimental sepsis led to a systemic cytokine storm resulting in the formation of excessive amounts of both pro-inflammatory cytokines (TNF-α, IL-1β, IL-17 and IL-6) and the anti-inflammatory cytokine IL-10. The systemic inflammatory response was accompanied by high plasma levels of ALT, AST (liver injury), creatinine, (renal dysfunction) and lactate, as well as a high, clinical severity score. All parameters were attenuated following PF271 administration. Experimental sepsis induced an overactivation of FAK and Pyk2 in liver and kidney, which was associated to p38 MAPK activation, leading to increased expression/activation of several pro-inflammatory markers, including the NLRP3 inflammasome complex, the adhesion molecules ICAM-1, VCAM-1 and E-selectin and the enzyme NOS-2 and myeloperoxidase. Treatment with PF271 inhibited FAK-Pyk2 activation, thus blunting the inflammatory abnormalities orchestrated by sepsis. Finally, PF271 significantly prolonged the survival of mice subjected to CLP-sepsis. Taken together, our data show for the first time that the FAK-Pyk2 pathway contributes to sepsis-induced inflammation and organ injury/dysfunction and that the pharmacological modulation of this pathway may represents a new strategy for the treatment of sepsis.
脓毒症和脓毒性休克与高死亡率相关,被认为是主要的公共卫生关注问题之一。脓毒症的发作被认为是一种过度炎症状态,导致器官衰竭和死亡。最近的研究结果表明,两种非受体蛋白酪氨酸激酶,即粘着斑激酶(FAK)和富含脯氨酸的酪氨酸激酶 2(Pyk2),在与子宫内膜异位症、癌症、动脉粥样硬化和哮喘相关的炎症中可能发挥作用。在这里,我们研究了 FAK-Pyk2 在脓毒症发病机制中的作用,以及通过给予强效可逆的 FAK 和 Pyk2 双重抑制剂 PF562271(PF271)在盲肠结扎和穿刺(CLP)诱导的脓毒症小鼠模型中对该途径进行药理学调节的潜在有益作用。五个月大的雄性 C57BL/6 小鼠接受 CLP 或假手术,手术后一小时,小鼠被随机分配接受 PF271(25mg/kg,sc)或载体。手术后 24 小时,收集器官和血浆进行分析。在另一组小鼠中,在随后的 5 天内每 12 小时评估一次存活率。实验性脓毒症导致全身细胞因子风暴,导致过量的促炎细胞因子(TNF-α、IL-1β、IL-17 和 IL-6)和抗炎细胞因子 IL-10 的形成。全身炎症反应伴随着高血浆 ALT、AST(肝损伤)、肌酐(肾功能障碍)和乳酸水平,以及高临床严重程度评分。PF271 给药后,所有参数均减弱。实验性脓毒症在肝和肾中引起 FAK 和 Pyk2 的过度激活,这与 p38 MAPK 的激活有关,导致几种促炎标志物的表达/激活增加,包括 NLRP3 炎性体复合物、粘附分子 ICAM-1、VCAM-1 和 E-选择素以及酶 NOS-2 和髓过氧化物酶。PF271 治疗抑制了 FAK-Pyk2 的激活,从而减轻了脓毒症引起的炎症异常。最后,PF271 显著延长了接受 CLP-脓毒症的小鼠的存活时间。总之,我们的数据首次表明,FAK-Pyk2 途径参与了脓毒症引起的炎症和器官损伤/功能障碍,并且该途径的药理学调节可能代表脓毒症治疗的新策略。