权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Potential usefulness of siRNAs for gene therapy ofsepsis-induced multiple organ failure

siRNA 在脓毒症引起的多器官衰竭基因治疗中的潜在用途

基本信息

批准号：
18590233
负责人：
HATTORI Yuichi
金额：
$ 2.46万
依托单位：
University of Toyama
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590233/
关键词：
Sepsis RNA interference Apoptosis Endothelial cells Acute lung injury Caspase-8 -3 FADD Small interfering RNA(siRNA)RNA千渉短鎖干渉RNA (siRNA)内皮由来NO合成酵素 caspase-3

项目摘要

Rationale: Not only better understanding of the molecular mechanisms involved in the pathogenesis of sepsis and its resultant organ failure, but also new therapeutic approaches and targets are urgently needed. Accumulating evidence suggests that apoptosis plays an important role in the pathophysiology of sepsis, and apoptosis may be potentially detrimental in septic acute lung injury (ALI). Also, vascular endothelial cell apoptosis may play a role in the pathogenesis of the septic syndrome.Objectives: We tested the hypothesis that systemic administration of small interfering RNA (siRNA) targeting Fas-associated death domain (FADD), which recruits procaspase-8 into the death-inducing signaling complex, may be protective in septic ALI and mortality We also evaluated the therapeutic efficacy of caspase-8/caspase-3 siRNAs in a murine model of polymicrobial endotoxic shock.Methods: Polymicrobial sepsis was induced by cecal ligation and puncture (CLP) in BALB/c mice. In vivo delivery of siRN … More As was performed by using a transfection reagent (Lipofectamine^(TM) RNAiMAX) at 10 hours after CLP. As a negative control, animals received non-sense (scrambled) siRNA.Measurements and Main Results: In CLP-induced septic mice, surface expression of death receptors were up-regulated and FADD was highly expressed and activated. DNA fragmentation ladder and TUNEL (transferase-mediated dUTP nick end labeling) assays showed that treatment with siRNAs suppressed apoptosis induction in lungs and vascular tissues following sepsis. Moreover, these siRNA treatments prevented the development of ALI in CLP mice, as indicated by great improvements of blood-gas derangements, histologic lung damage, and increased pulmonary inflammatory cells. Furthermore, hematoxylin/eosin-stained sections of septic aorta displayed partial detachment of endothelial cells from the basal membrane. These histopathologic changes in endothelial cells were drastically prevented by systemic treatment with FADD siRNA or caspase-8/-3 siRNAs. Finally, administration of FADD siRNA or caspase-8/-3 dramatically improved the survival of CLP mice.Conclusions: These results indicate the pathophysiological significance of the death receptor apoptotic pathway, including FADD, in septic ALI and the potential usefulness of FADD siRNA for gene therapy of the septic syndrome. In addition, gene silencing of caspase-8 and caspase-3 with siRNAs provided profound protection against polymicrobial endotoxic shock. The prevention of vascular endothelial cell apoptosis appears to be, at least in part, responsible for their beneficial effects in endotoxic shock. Less

理论基础：不仅需要更好地了解脓毒症及其导致的器官衰竭的发病机制，而且迫切需要新的治疗方法和靶点。越来越多的证据表明细胞凋亡在脓毒症的病理生理过程中起着重要作用，细胞凋亡在脓毒症急性肺损伤(ALI)中可能具有潜在的危害作用。此外，血管内皮细胞的凋亡可能在脓毒症综合征的发病机制中起作用。目的：我们验证了全身应用针对Fas相关死亡结构域(FADD)的小干扰RNA(SiRNA)可能对感染性ALI和死亡率具有保护作用的假说。我们还评价了caspase-8/caspase-3 siRNA在多菌性内毒素休克小鼠模型中的治疗效果。SiRN…的体内递送在CLP后10h使用转染剂(Lipofetamine^(TM)RNAiMAX)进行更多的检查。实验方法和主要结果：在CLP诱导的脓毒症小鼠中，死亡受体的表面表达上调，FADD高表达和激活。DNA梯状条带和TUNEL(转移酶介导的dUTP缺口末端标记)分析表明，siRNAs治疗可抑制脓毒症后肺和血管组织的细胞凋亡诱导。此外，这些siRNA治疗阻止了CLP小鼠ALI的发展，表现为血气紊乱、组织学肺损伤和肺炎性细胞增加的显著改善。此外，败血症大动脉苏木精/伊红染色切片显示内皮细胞部分脱离基底膜。用FADD siRNA或caspase-8/-3 siRNA系统治疗后，内皮细胞的这些组织病理学改变被显著地预防。结论：包括FADD在内的死亡受体凋亡通路在脓毒症ALI中具有重要的病理生理学意义，FADD siRNA在败血症综合征的基因治疗中具有潜在的应用价值。此外，通过siRNAs沉默caspase-8和caspase-3的基因，可以有效地对抗多菌素性内毒素休克。防止血管内皮细胞凋亡似乎至少在一定程度上是它们对内毒素休克的有益作用的原因。较少