Center for Immunology and Inflammation, The Feinstein Institutes for Medical Research, Manhasset, NY.
Department of Surgery, Zucker School of Medicine at Hofstra/Northwell, Manhasset, NY.
J Immunol. 2023 Oct 1;211(7):1144-1153. doi: 10.4049/jimmunol.2300228.
Sepsis is an infectious inflammatory disease that often results in acute lung injury (ALI). Cold-inducible RNA-binding protein (CIRP) is an intracellular RNA chaperon that binds to mRNA's poly(A) tail. However, CIRP can be released in sepsis, and extracellular CIRP (eCIRP) is a damage-associated molecular pattern, exaggerating inflammation, ALI, and mortality. In this study, we developed an engineered poly(A) mRNA mimic, AAAAAAAAAAAA, named A12, with 2'-O-methyl ribose modification and terminal phosphorothioate linkages to protect it from RNase degradation, exhibiting an increased half-life. A12 selectively and strongly interacted with the RNA-binding motif of eCIRP, thereby preventing eCIRP's binding to its receptor, TLR4. In vitro treatment with A12 significantly decreased eCIRP-induced macrophage MAPK and NF-κB activation and inflammatory transcription factor upregulation. A12 also attenuated proinflammatory cytokine production induced by eCIRP in vitro and in vivo in macrophages and mice, respectively. We revealed that treating cecal ligation and puncture-induced sepsis with A12 significantly reduced serum organ injury markers and cytokine levels and ALI, and it decreased bacterial loads in the blood and peritoneal fluid, ultimately improving their survival. Thus, A12's ability to attenuate the clinical models of sepsis sheds lights on inflammatory disease pathophysiology and prevention of the disease progress.
脓毒症是一种感染性炎症性疾病，常导致急性肺损伤（ALI）。冷诱导 RNA 结合蛋白（CIRP）是一种细胞内 RNA 伴侣，可与 mRNA 的 poly(A)尾结合。然而，CIRP 可在脓毒症中释放，细胞外 CIRP（eCIRP）是一种损伤相关分子模式，可加剧炎症、ALI 和死亡率。在这项研究中，我们开发了一种工程化的 poly(A)mRNA 模拟物，名为 A12，它具有 2'-O-甲基核糖修饰和末端硫代磷酸酯键，以防止其被 RNase 降解，从而延长半衰期。A12 选择性且强烈地与 eCIRP 的 RNA 结合基序相互作用，从而阻止 eCIRP 与其受体 TLR4 结合。体外用 A12 处理可显著降低 eCIRP 诱导的巨噬细胞 MAPK 和 NF-κB 激活以及炎症转录因子的上调。A12 还可分别减弱 eCIRP 在体外和体内诱导的巨噬细胞和小鼠促炎细胞因子的产生。我们揭示，用 A12 治疗盲肠结扎和穿刺诱导的脓毒症可显著降低血清器官损伤标志物和细胞因子水平以及 ALI，并降低血液和腹腔液中的细菌负荷，最终提高其生存率。因此，A12 减轻脓毒症临床模型的能力阐明了炎症性疾病的病理生理学和疾病进展的预防。
