Supraphysiologic Shear Stresses Associated with Cardiopulmonary Bypass are Sufficient to Activate RIKP3 Signaling

与心肺绕道相关的超生理剪切应力足以激活 RIKP3 信号传导

基本信息

批准号：
10446535
负责人：
VISHAL NIGAM
金额：
$ 82.98万
依托单位：
SEATTLE CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-11 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10446535
关键词：
Adrenal Cortex Hormones Animal Model Biomechanics Blood Blood Cells Bypass Calcium Calcium Signaling Cardiac Surgery procedures Cardiopulmonary Bypass Cell Death Cells Cellular Structures Cessation of life Childhood Clinical Congenital Heart Defects Critical Pathways Data Drug usage Exposure to Functional disorder Gene Expression Goals Health Care Costs Impairment In Vitro Infant Inflammation Inflammatory Inflammatory Response Knowledge Leukocytes Lytic Mediating Mediator of activation protein Membrane Molecular Morbidity - disease rate Myeloid Cells Neonatal Operative Surgical Procedures Organ Outcome Pathway interactions Patients Pattern Phenotype Phosphorylation Phosphotransferases Physiological Plasma Play Process Protein-Serine-Threonine Kinases Publishing RIPK3 gene Research Role STIM1 gene Signal Pathway Signal Transduction Techniques Temperature Testing Tissues Translating base cell cortex cytokine effectiveness evaluation experience experimental study improved outcome in vivo inhibitor insight migration monocyte mortality neonatal patient novel novel strategies palliation porcine model preclinical development receptor repaired response shear stress small molecule systemic inflammatory response treatment strategy

项目摘要

The vast majority of pediatric open-heart surgeries require the patient to be supported by cardiopulmonary bypass (CPB). Exposure to CPB causes systemic inflammation and resultant multi-organ dysfunction. Post-CPB inflammation is believed to be caused by exposure of the patient’s blood cells to the plastic tubing of the CPB circuit and unphysiological high shear stress. However, the mechanisms underlying this process are unclear. Our long-term goal is to understand how these insults contribute to post-CPB inflammation and translate this knowledge into novel treatment strategies. The scientific premise for this project is that Receptor-Interacting serine/threonine-Protein Kinase 3 (RIPK3) signaling is a novel mediator of CPB associated inflammation that can be targeted to improve outcomes for neonatal cardiac surgery patients. Data from neonatal CBP patients, a large animal model of CPB, and in vitro experiments form the basis of our scientific premise. We recently published that RIPK3 and necroptosis play a role in post-CPB inflammation. Our data demonstrate that RIPK3 plays a role in post-CPB inflammation. We established that supraphysiologic shear stresses present during CPB are sufficient to activate RIPK3 signaling in vitro and in a piglet model of CPB. Mechanistically, we found that shear stress-initiated calcium signaling pathways are critical to the activation of monocytic cells. We have identified specific pathways that can be targeted with small molecules to reduce CPB-activation of RIPK3 with goal of reducing systemic inflammatory response and organ dysfunction. We hypothesize that CPB-associated shear stress activates RIPK3 mediated inflammation. The objectives of this proposal are 1) to determine how CPB activates RIPK3 signaling, 2) elucidate how RIPK3 signaling contributes to CPB-associated inflammation/organ dysfunction, and 3) determine if blocking RIPK3 signaling is sufficient to reduce CPB- associated inflammation and organ dysfunction. Our approach will consist of two specific aims: Aim 1. Determine how shear stress activates RIPK3 signaling in circulating myeloid cells. We postulate that supraphysiologic shear stress is sufficient to activate RIPK3 signaling. We will characterize the shear stress thresholds and molecular mechanism responsible for RIPK3 activation during CPB with a focus on the roles that the cell cortex, calcium signaling cascade, and shear responsive kinases play in this response. Aim 2. Demonstrate that RIPK3 signaling mediates CPB-associated inflammation and organ dysfunction. We postulate that RIPK3 signaling is required for the inflammatory response to CPB. RIPK3 can help propagate inflammation via necroptosis, the release of cytokines, and leukocyte migration. We will perform in vitro and in vivo experiments to demonstrate that targeting RIPK3 signaling reduces CPB-associated inflammation. This research is novel and significant – elucidating how CPB activates RIPK3 signaling and necroptosis could enable a new treatment paradigm for CPB patients, improve outcomes, and reduce healthcare costs, since the proposed signaling pathways can be targeted by small molecules in clinical use or pre-clinical development.

绝大多数儿科心脏直视手术都需要患者获得心肺支持暴露于体外循环（CPB）会导致全身炎症并导致体外循环后的多器官功能障碍。炎症被认为是由于患者的血细胞暴露于 CPB 的塑料管引起的然而，这一过程的机制尚不清楚。我们的长期目标是了解这些侮辱如何导致 CPB 后炎症，并将其转化为该项目的科学前提是受体相互作用。丝氨酸/苏氨酸蛋白激酶 3 (RIPK3) 信号传导是 CPB 相关炎症的新型介质，可以有针对性地改善新生儿心脏手术患者的结果，来自新生儿 CBP 患者的数据。 CPB 的大型动物模型和体外实验构成了我们最近的科学前提的基础。发表了 RIPK3 和坏死性凋亡在 CPB 后炎症中发挥作用的研究，我们的数据表明 RIPK3。我们确定在 CPB 期间存在超生理剪切应力。足以在体外和 CPB 仔猪模型中激活 RIPK3 信号传导，我们发现剪切应力引发的钙信号通路对于单核细胞的激活至关重要。确定了可以用小分子靶向的特定途径，以减少 RIPK3 的 CPB 激活减少全身炎症反应和器官功能障碍的目标。剪切应力激活 RIPK3 介导的炎症该提案的目标是 1) 确定如何激活。 CPB 激活 RIPK3 信号传导，2) 阐明 RIPK3 信号传导如何促进 CPB 相关炎症/器官功能障碍，3) 确定阻断 RIPK3 信号传导是否足以减少 CPB- 我们的方法将包括两个具体目标：目标 1. 确定剪切应力如何激活循环骨髓细胞中的 RIPK3 信号传导。我们假设超生理剪切应力足以激活 RIPK3 信号传导。重点关注 CPB 期间 RIPK3 激活的剪切应力阈值和分子机制细胞皮质、钙信号级联和剪切响应激酶在此反应中发挥的作用。目标 2. 证明 RIPK3 信号传导介导 CPB 相关炎症和器官功能障碍。我们假设 RIPK3 信号传导是 CPB 炎症反应所必需的，可以帮助传播。通过坏死性凋亡、细胞因子的释放和白细胞迁移产生的炎症我们将在体外和体内进行。体内实验证明靶向 RIPK3 信号传导可减少 CPB 相关炎症。这项研究新颖且意义重大——阐明 CPB 如何激活 RIPK3 信号传导和坏死性凋亡为 CPB 患者提供新的治疗模式，改善结果并降低医疗成本，因为所提出的信号通路可以在临床使用或临床前开发中被小分子靶向。