Prospective Evaluation of PLatelets Acetyl-CoA Carboxylase Phosphorylation State in SEPtic Shock Patients. Impact of Platelets Metabolism to Inflammatory Response.
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 150
- 试验地点
- 2
- 主要终点
- Platelets Acetyl-CoA Carboxylase phosphorylation rate
研究概览
简要总结
Knowing the dramatic increase in thrombin generation during sepsis, our research hypothesis is that AMPK-induced ACC phosphorylation in platelets is increased and that this might modulate platelets metabolism and more particularly platelets inflammatory mediators content, coming from AA and lipids.
详细描述
Background
Sepsis, a systemic inflammatory reaction occurring in response to an infection, is a major public health problem and is recognized as the leading cause of death in the intensive care unit (ICU). This severe inflammatory reaction causes endothelial activation and dysfunction. This phenomenon is closely associated with the activation of coagulation cascade via exposure of tissue factor (TF) at endothelial cell surface. TF serves as an anchor for factors of coagulation VII and X allowing their activation. This results in thrombin (IIa) generation and fibrin production.
Thrombin is also a potent platelet agonist (activator) resulting in platelet activation and consumption. The pro-coagulant activity of activated platelets during sepsis enhances the formation of micro-thrombosis that impair organ perfusion and may lead to death. Thrombocytopenia is common and is a strong negative prognostic marker in septic shock patients.
Beyond coagulation, platelets are known to play a key role in inflammation and immune response. Platelets activation can be initiated by inflamed endothelium or circulatory inflammatory cytokines, which in turn, modulates the inflammatory and thrombotic response. Activated platelets interact with immune cells, like neutrophils. Furthermore, platelets can act as microbial "sensors" by expressing members of Toll-Like Receptors (TLRs) family, binding ligands from several infectious agents. Recent data highlight the interplay between platelets and neutrophils extracellular traps (NETs). Platelets facilitate NETs formation and, inversely, NETS activate platelets. NETs connect platelets, thrombosis, immune response and inflammation and therefore are of particular interest in the septic context.
Platelets express Acetyl-CoA carboxylase 1 (ACC1), which is the first step enzyme of lipogenesis. ACC Phosphorylation on Serine 79 by AMP-activated protein kinase (AMPK) leads to its inhibition and AMPK is activated by thrombin. We demonstrated that AMPK-ACC axis controls platelets phospholipids content, which influence TXA2 and dense granule release and, in turn, thrombus formation. TXA2 is generated from phospholipids containing arachidonic acid (AA). Alternatively, AA can be metabolized by lipooxygenase (LOX) pathway producing lipoxins (LX) and resolvin, which are rather involved in the resolution of inflammation. The relation between AMPK-ACC signalling and lipooxygenase pathway has never been investigated.
研究设计
- 研究类型
- Interventional
- 分配方式
- Non Randomized
- 干预模型
- Parallel
- 主要目的
- Basic Science
- 盲法
- None
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Informed consent of patients, their relatives or independent physician
- •Septic shock defined as a sepsis with vasopressor therapy needed to elevate MAP ≥65 mmHg, and lactate > 2 mmol/L, despite adequate fluid resuscitation of 30mL/kg of intravenous crystalloid within 6 hours. Inclusion within 48 hours of ICU admission.
- •Covid-19 patients with ARDS and PaO2/FiO2 <
- •Inclusion within 5 days after ICU admission.
排除标准
- •Patients on therapeutic anticoagulation therapy (oral or parenteral) including heparins, fondaparinux, vitamin K antagonist, novel oral anticoagulants, for any reasons DESPITE therapeutic anticoagulation as a treatment for Covid-19 patients.
- •Recent (less than 1 month) chemotherapy
- •Active inflammatory disease
- •Haemophilia and other coagulopathy
- •Previous history of thrombocytopenia (<100 000 platelets/mm3)
- •Cirrhosis (Child Plug > A)
- •Recent (less than 48 hours) major surgery
- •Exclusion criteria for Covid-19 patients:
- •- bacterial co-infection
结局指标
主要结局
Platelets Acetyl-CoA Carboxylase phosphorylation rate
时间窗: At the time of inclusion
ACC phosphorylation on Ser79 (phosphoACC) in platelets of patients will be assessed using western blotting. Results will be expressed in arbitrary units (A.U). A signal above 0.5 A.U. has already been shown to be above 2 standard deviation in a healthy population.
次要结局
- Mortality rate(30 days and 1-year follow-up)
- Tubulin acetylation rate(At the time of inclusion)
- Platelets activation(At the time of inclusion)
- Neutrophils extracellular trap formation(At the time of inclusion)
- Respiratory failure(At the time of inclusion and through study completion up to day 30)
- Total platelets lipid content and composition(At the time of inclusion)
- Platelet function assessment(At the time of inclusion)
- Sepsis severity(At the time of inclusion)
- Thrombin generation marker rate(At the time of inclusion)