Proteasome activation to protect the white matter in neonatal hypoxic-ischemic encephalopathy.

蛋白酶体激活保护新生儿缺氧缺血性脑病中的白质。

基本信息

批准号：
10223450
负责人：
Jennifer Kim Lee
金额：
$ 55.46万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10223450
关键词：
3-Dimensional Acute Adjuvant Adjuvant Therapy Affect Apoptosis Asphyxia Neonatorum Atrophic Axon Bioavailable Biochemistry Biology Blood gas Brain Brain Hypoxia-Ischemia Brain Injuries Cell Culture Techniques Cell Death Cell Differentiation process Cell Lineage Cell Maturation Cells Cellular biology Cerebrum Clinical Clinical Research Cognition Cognitive Cognitive deficits Data Dependovirus Dose Electrolytes Electron Microscopy Exhibits Failure Family suidae Genetic Glucose Goals Health Human Impairment Infant Injury Intravenous Magnetic Resonance Imaging Mediating Mediator of activation protein Methods Modeling Monitor Myelin Myelin Proteins Myelin Sheath Neonatal Neonatal Intensive Care Nervous System Trauma Neurocognitive Neurocognitive Deficit Neurologic Neurological outcome Neurons Newborn Infant Oligodendroglia Olives - dietary Outcome Oxidative Stress Oxides Oxygen Patients Pharmaceutical Preparations Physiological Processes Post-Translational Protein Processing Property Prosencephalon Proteasome Inhibition Proteasome Inhibitor Protein Biosynthesis Proteins Quality Control Recovery Regimen Regulation Resistance Rewarming Risk Role Sedation procedure Serotyping Small Interfering RNA Term Birth Testing Therapeutic Ubiquitination Ventilator Virus axon injury behavior test clinical care clinically relevant cohort cytokine cytotoxic deprivation disability experimental study functional outcomes hemodynamics improved knock-down mortality risk motor impairment multicatalytic endopeptidase complex myelination natural hypothermia neonatal death neonatal hypoxic-ischemic brain injury neonate neuropathology novel oligodendrocyte precursor oxidation oxidative damage porcine model postnatal precursor cell prenatal preservation protein degradation side effect small hairpin RNA small molecule small molecule inhibitor therapeutic target white matter white matter injury

项目摘要

Neonatal hypoxic-ischemic encephalopathy (HIE) from birth asphyxia causes persistent and severe neurologic disabilities, even in patients who receive therapeutic hypothermia. We found in clinical studies that white matter injury on MRI persists after hypothermic treatment. Thus, hypothermia is not fully protective. White matter injury is a prominent yet understudied component of the neurologic disabilities observed in neonates who receive hypothermia for HIE. Therapeutic adjuncts that protect the white matter might reduce the risk of permanent neurologic injury in HIE. Hypoxia-ischemia (HI) in neonatal pig, which has human-like white matter tracts, produces brain damage similar to that of full-term human newborns with HIE, including the white matter injuries observed in clinical studies. Our model includes clinically relevant whole-body hypothermia, rewarming at 0.5°C/h, sedation, continuous hemodynamic monitoring, ventilator support, and correction of blood gas and electrolyte abnormalities to mimic clinical neonatal intensive care. Preliminary data suggest that insufficient proteasome function mediates persistent white matter injury after HI and hypothermia. We postulate that white matter proteasome insufficiency causes a failure to clear oxidatively damaged proteins, causing oligodendrocyte apoptosis, potential disruption of oligodendrocyte precursor maturation, myelin and axonal injury, and white matter volume loss after HI and hypothermia. We will elucidate the proteasome’s role in white matter injury after whole-body HI and overnight hypothermia in neonatal swine. White matter injury and oligodendrocyte biology will be studied with neuropathology (including oligodendrocyte precursor maturation, stereology, cell death, and electron microscopy) and biochemistry (including protein post-translational modification and proteasome composition and activity) through 1 month recovery after HI. T- maze neurocognitive behavior testing with neuropathology correlation will provide a functional outcome. We developed new methods to genetically modulate proteasome activity in distinct, targeted regions of white matter in neonatal pig forebrain using virus-mediated enforced expression of a proteasome activator subunit or proteasome inhibition with short hairpin small interfering RNA. We will also use a small molecule proteasome inhibitor to determine whether proteasome inhibition aggravates white matter injury. Moreover, we will test the potential of the drug oleuropein to protect white matter. Oleuropein is a readily bioavailable compound with proteasome activating properties and few clinical side effects. An intravenous oleuropein dosing regimen will be used that protects oligodendrocytes and myelin, increases proteasome expression, and promotes clearance of oxidized proteins after HI and hypothermia. We will identify whether oleuropein acts on the standard proteasome or the immunoproteasome. Cultured human oligodendrocyte experiments will validate the proteasome as a therapeutic target and oleuropein’s actions after oxygen glucose deprivation. This project will advance the neonatal HI and cell biology fields by investigating novel mechanisms by which proteasome insufficiency mediates hypothermia-resistant injury in white matter. We will discover whether proteasome activation is a relevant therapeutic adjunct to hypothermia to protect white matter and improve neurologic outcomes in HIE.

新生儿缺氧性 - 缺血性脑病（HIE）从出生窒息引起持续和严重的神经系统残疾，即使在接受治疗性体温过低的患者中也是如此。我们在临床研究中发现白质损伤在低温治疗后，MRI持续存在。这是体温过低的。白质伤害是突出的然而，在接受HIE体温过低的新生儿中观察到的神经系统疾病的成分。保护白质的治疗辅助物可能会降低HIE永久神经系统损伤的风险。新生儿猪中的缺氧 - 缺血（HI），具有人类的白质区，会产生类似的脑损伤与HIE的完整人类新生儿，包括在临床研究中观察到的白质损伤。我们的模型包括临床上相关的全身低温，在0.5°C/h时重新传输，镇静，连续血流动力学对模仿临床新生儿的血液和电解质异常的监测，通风支持和纠正重症监护。初步数据表明，蛋白酶体功能不足培养持续性白质损伤在嗨和体温过低之后。我们假设白质蛋白酶体不足会导致无法氧化清除受损的蛋白质，导致少突胶质细胞凋亡，潜在的少突胶质细胞前体成熟的破坏， HI和体温过低后的髓磷脂和轴突损伤以及白质体积损失。我们将阐明蛋白酶体在全身HI和隔夜体温过低后在白质损伤中的作用新生儿猪。白质损伤和少突胶质细胞生物学将研究神经病理学（包括少突胶质细胞前体的成熟，立体学，细胞死亡和电子显微镜）和生物化学（包括蛋白质后翻译后修饰，蛋白酶体组成和活性），直到1个月后恢复。 t- 与神经病理相关性的迷宫神经认知行为测试将提供功能结果。我们开发了在新生儿猪中不同的，靶向的白质区域中遗传调节蛋白酶体活性的新方法前脑使用病毒介导的蛋白酶体激活剂亚基或蛋白酶体抑制的强化表达短发夹小干扰RNA。我们还将使用小分子蛋白酶体抑制剂来确定是否是否蛋白酶体抑制会加剧白质损伤。此外，我们将测试药物蛋白蛋白保护的潜力白质。 Oleuropein是一种易于生物利用化合物，具有蛋白酶体激活特性，临床方面很少效果。将使用静脉注射油蛋白剂量方案，以保护少突胶质细胞和髓磷脂，增加蛋白酶体的表达，并促进HI和低温后氧化蛋白的清除。我们将确定是否油蛋白作用于标准蛋白酶体或免疫蛋白酶体。培养的人类少突胶质细胞实验将验证蛋白酶体为氧葡萄糖剥夺后的治疗靶标和油蛋白酶的作用。这项目将通过研究蛋白酶体的新机制来推动新生儿HI和细胞生物学领域功能不全介导白质中的抗性低温损伤。我们将发现蛋白酶体激活是否是体温过低的相关治疗辅助性保护白质并改善了HIE的神经系统效果。