Inhibition of necroptosis during inflamm-aging and pneumonia

抑制炎症老化和肺炎期间的坏死性凋亡

• 批准号: 9248088
• 负责人: Carlos J Orihuela
• 金额: $ 18.38万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9248088
• 关键词: Adipose tissue; Adult Respiratory Distress Syndrome; Aftercare; Age; Age-Months; Age-Years; Aging; Alveolar Macrophages; Alveolus; Animals; Apolipoprotein E; Arterial Fatty Streak; B-Lymphocytes; Bacterial Pneumonia; Blood; Bone Marrow; Brain; Cardiovascular Diseases; Case Fatality Rates; Cause of Death; Cell Aging; Cell Death; Cell membrane; Cells; Characteristics; Chronic; Chronic Disease; Disease; Elderly; Exposure to; Extravasation; Flow Cytometry; Functional disorder; Gait; Health; Health Status; Heart; Heterozygote; High Fat Diet; Human; Immune; Individual; Infection; Inflammation; Inflammatory; Injury; Kidney; Knockout Mice; Knowledge; Laboratories; Lead; Learning; Liver; Longevity; Lower Respiratory Tract Infection; Lung; MAP Kinase Gene; Measures; Mediating; Mitochondria; Morbidity - disease rate; Mus; Outcome; Oxidative Stress; Pathology; Pathway interactions; Play; Pneumococcal Pneumonia; Pneumonia; Predisposition; Preventive Intervention; Protein Kinase; RIPK1 gene; RIPK3 gene; Reactive Oxygen Species; Reporting; Role; Serum; Severities; Severity of illness; Spleen; Streptococcus pneumoniae; Streptococcus pneumoniae plY protein; T-Lymphocyte; Testing; Therapeutic Intervention; Thymus Gland; Tissues; Toxin; Translational Research; Virus Diseases; Visceral; age related; aged; alveolar bone; cell injury; cytokine; experience; functional decline; improved; inflammatory marker; inhibitor/antagonist; juvenile animal; lung injury; macrophage; mitochondrial dysfunction; monocyte; muscle strength; novel; pathogen; research study; response; sarcopenia; vascular inflammation

Necroptosis is a newly discovered cell death pathway. Regulated by receptor-interacting protein kinase (RIP)1 and RIP3, necroptosis involves the purposeful disruption of the cell membrane by the effecter molecule MLKL. Necroptosis is inflammatory due to the release of cytoplasmic components that act as alarmins. Necroptosis is thought to amplify and promote inflammatory circles that contribute to chronic disease. What role necroptosis plays during aging is unknown. For example, does it contribute to inflamm-aging? Pore-forming toxins produced by Streptococcus pneumoniae and other airway pathogens have been shown to trigger lung cell necroptosis and this is responsible for much of the injury that is observed. The elderly are in particular vulnerable to pneumonia, with lower respiratory tract infections being the 4th leading cause of death in those ≥65 years of age. Thus, inhibition of necroptosis is potential way to protect vulnerable individuals, such as the elderly, from the lung damage that occurs during bacterial pneumonia. We hypothesize that cell death by necroptosis increases with advanced age and this contributes to inflamm- aging. Also, that blocking necroptosis can protect the elderly from lung injury during pneumonia. Herein, we will leverage our expertise on aging, necroptosis, and bacterial pneumonia to test these hypotheses and improve our understanding of necroptosis and its impact during aging. We will: AIM 1. Determine the role of necroptosis on inflamm-aging and age-related decline in function. We have mice deficient in RIP3 and MLKL that cannot undergo necroptosis. We will compare pro-inflammatory cytokine and alarmin profiles in serum from WT, heterozygote, and KO mice at 6, 14, and 24 months of age. We will also examine aged WT mice treated for 7 days with two different necroptosis inhibitors. For all mice, we will examine the activation status of NFkB and MAPK in the thymus, lungs, heart, spleen, liver, kidney, brain, and visceral adipose by western and in immune cells (monocytes, B cells, T cells) by flow cytometry. We will also longitudinally measure activity, gait, and muscle strength to learn if blocking necroptosis impacts health status. AIM 2. Determine if blocking necroptosis protects aged animals against pneumonia. Alveolar macrophages and bone marrow derived macrophages from 6, 14 and 24 month old mice will be tested for their propensity to undergo necroptosis following exposure to the pore-forming toxin pneumolysin. Different aged WT, heterozygote, and MLKL KO mice, along with WT mice treated with necroptosis inhibitors will be intratracheally challenged with pneumolysin or infected with S. pneumoniae. Lung damage and disease severity will be assessed by measuring inflammatory cytokines, pathology, and bacterial burden, respectively. This proposal is in response to PAR-14-191:T1 Translational Research: Novel Interventions for Prevention and Treatment of Age-Related Conditions. We will determine the contribution of necroptosis to inflamm-aging and learn if blocking necroptosis protects against inflamm-aging related decline and pneumonia.

坏死性凋亡是一种新发现的细胞死亡途径。受受体相互作用蛋白激酶（RIP）1调节 和RIP3，坏死性凋亡涉及效应分子MLKL有目的地破坏细胞膜。 坏死性凋亡是由于释放细胞质成分作为警报素引起的炎症。坏死性上睑下垂是 被认为放大和促进炎症循环，导致慢性疾病。坏死性凋亡有什么作用 衰老的过程是未知的。例如，它是否有助于炎症老化？ 肺炎链球菌和其他呼吸道病原体产生的成孔毒素已被证明， 引发肺细胞坏死性凋亡，这是造成所观察到的大部分损伤的原因。老年人在 特别容易感染肺炎，下呼吸道感染是第四大死因 在≥ 65岁的患者中。因此，抑制坏死性凋亡是保护易受伤害的人的潜在途径。 例如老年人，在细菌性肺炎期间发生的肺损伤。 我们假设，随着年龄的增长，坏死性凋亡引起的细胞死亡增加，这有助于炎症反应。 衰老此外，阻断坏死性凋亡可以保护老年人在肺炎期间免受肺损伤。在此，我们将 利用我们在衰老、坏死性凋亡和细菌性肺炎方面的专业知识来验证这些假设， 我们对坏死性凋亡及其在衰老过程中的影响的理解。我们将： AIM 1.确定坏死性凋亡在炎症老化和年龄相关功能下降中的作用。我们有 RIP3和MLKL缺陷小鼠不能发生坏死性凋亡。我们将比较促炎细胞因子 和6、14和24月龄的WT、杂合子和KO小鼠的血清中的alarmin谱。我们将 还检查了用两种不同的坏死性凋亡抑制剂处理7天的老年WT小鼠。对于所有的老鼠，我们将 检测胸腺、肺、心脏、脾、肝、肾、脑中NF κ B和MAPK的活化状态， 免疫细胞（单核细胞、B细胞、T细胞）中通过Western和流式细胞术检测。我们还将 纵向测量活动、步态和肌肉力量，以了解阻断坏死性凋亡是否影响健康状况。 AIM 2.确定阻断坏死性凋亡是否能保护老年动物免受肺炎的侵害。肺泡 将测试来自6、14和24月龄小鼠的巨噬细胞和骨髓衍生的巨噬细胞的 在暴露于成孔毒素肺炎球菌溶血素后发生坏死性凋亡的倾向。不同年龄 WT、杂合子和MLKL KO小鼠，沿着用坏死性凋亡抑制剂处理的WT小鼠， 用肺炎链球菌溶血素进行肠内攻毒或感染S.肺炎。肺损伤和疾病 通过分别测量炎性细胞因子、病理学和细菌负荷来评估严重程度。 本提案是对PAR-14 - 191：T1转化研究：新的干预措施， 预防和治疗与糖尿病相关的疾病。我们将确定坏死性凋亡对 了解阻断坏死性凋亡是否能预防炎症-衰老相关的衰退和肺炎。