Probing Role of Tetrahydrobiopterin in Cerebral Palsy by Using Transgenic Rabbits

利用转基因兔探讨四氢生物蝶呤在脑瘫中的作用

基本信息

批准号：
10312139
负责人：
SIDHARTHA TAN
金额：
$ 63.74万
依托单位：
WAYNE STATE 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：
2020-12-15 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10312139
关键词：
Abbreviations Abruptio Placentae Acute Animal Model Animals Behavior Birth Brain Brain Hypoxia-Ischemia Brain Injuries Brain region Breeding CRISPR/Cas technology Cell Therapy Cells Cerebral Palsy Cessation of life Child Childhood Congenital Disorders Critical Pathways Cues DHFR gene DNA Sequence Alteration Detection Development Diffusion Dihydrofolate Reductase Discipline of obstetrics Disease Early identification Enzymes Equilibrium Etiology Event Family suidae Fetus Flow Cytometry Functional disorder GTP Cyclohydrolase I Genes Genetic Genetic Engineering Genome High Pressure Liquid Chromatography Hospitals Human Human Genetics Hypoxia Inflammation Injury Institution Knock-in Knock-out Lead Life Link Lipopolysaccharides Magnetic Resonance Imaging Mammals Methods Modeling Molecular Motor Motor Manifestations Movement Disorders Muscle Hypertonia Mutation Nature Neurons Neurotransmitters Newborn Infant Nitric Oxide Nitric Oxide Synthase Nitric Oxide Synthase Type I Nitrogen Organ Oryctolagus cuniculus Outcome Oxidants Oxidoreductase Oxygen Parents Pathogenesis Pathway interactions Patients Pattern Perinatal Perinatal Brain Injury Periventricular Leukomalacia Phenotype Placental Insufficiency Predisposing Factor Predisposition Pregnancy Preventive Preventive treatment Productivity Reperfusion Therapy Rodent Model Role Sepiapterin reductase Severities Siblings Societies Superoxides Supplementation Testing Tetrahydrobiopterin deficiency Time Tissues Transgenic Organisms Variant antenatal antioxidant therapy behavioral phenotyping burden of illness cell injury cofactor dihydropteridine reductase effective therapy enzyme pathway fetal fetus hypoxia genetic manipulation human disease indexing innovation insight magnetic resonance imaging biomarker motor deficit motor disorder motor impairment neonatal hypoxic-ischemic brain injury neonate neuroprotection nonhuman primate oxidant stress perinatal brain postnatal prenatal rational design sepiapterin social tetrahydrobiopterin

项目摘要

PROJECT SUMMARY/ABSTRACT Children with movement disorders are a big burden to society. The burden of disease is very high because of the life-long consequences to the patient, caretakers, and social institutions. Currently there are no cures or preventative treatments for cerebral palsy (CP), as the mechanisms of disease remain poorly defined. Human mutations in key enzymatic pathways constitute genetic causes of childhood movement disorders. With the advent of transgenic rabbit models, a golden opportunity has arisen to study the pathogenetic mechanisms in brain leading to movement disorders, as rabbits are more likely to present with movement disorders mimicking that of humans. Rabbits are perinatal brain developers like humans. Mutations in enzymes of tetrahydrobiopterin pathway result in movement disorders. Tetrahydrobiopterin is an enzyme co-factor and its supplementation in congenital deficiency disorders ameliorates the movement disorder. Thus, there may be a critical role of tetrahydrobiopterin in the development of movement disorders, such as CP. We developed a knockout rabbit that introducing a specific mutation in one of the tetrahydrobiopterin synthesis enzymes, sepiapterin reductase. Following fetal hypoxia-ischemia, newborn rabbits present with hypertonia and difficulty with balance. Fetal rabbits showing low developmental tetrahydrobiopterin in discrete brain regions have a greater disposition to develop hypertonia. Magnetic resonance imaging (MRI) allows us to predict which fetuses will develop postnatal hypertonia. This advance allows the identification of early critical pathways causing hypertonia. Our objective is to elucidate molecular mechanisms of perinatal brain injury in human mutations causing childhood movement disorders, by decreasing tetrahydrobiopterin levels using a hetero- and homozygous knockout transgenic approach in the rabbit. The main question asked in this proposal is whether tetrahydrobiopterin in selective brain regions contributes to the development of motor disorders with a severity determined by an added prenatal insult such as hypoxia-ischemia or inflammation. Using genetic knockout of sepiapterin reductase, we can further lower the tetrahydrobiopterin levels in brain and investigate whether the resulting motor deficits are increased or that we need less degree of insult to achieve the same motor deficits. The first Aim determines whether an added fetal insult, hypoxia-ischemia or inflammation from lipopolysaccharide, enhances movement disorders in the sepiapterin het/homozygous reductase knockout rabbit. The second Aim will determine if neuronal or oligodendroglial injury explains the development of movement disorders in the knockout rabbit. We use innovative pre- and postnatal MRI biomarkers of hypertonia with tissue flow cytometry and high-performance liquid chromatography with electrochemical detection. By conducting a time-dependent, organ-specific and cell-specific pathogenetic study, we will obtain a comprehensive picture of the role of this cofactor in perinatal pathogenesis of movement disorders.

项目摘要/摘要运动障碍的儿童是社会的巨大负担。疾病负担很高，因为终身对患者，看护人和社会机构的后果。目前没有治疗方法或由于疾病的机制仍然很差，脑瘫（CP）的预防性处理。人类关键酶促途径中的突变构成了儿童运动障碍的遗传原因。与转基因兔模型的出现，已经出现了一个黄金的机会来研究中的致病机制大脑导致运动障碍，因为兔子更有可能出现模仿运动障碍人类。兔子是像人类一样的围产期脑开发人员。酶突变四氢无生蛋白途径导致运动障碍。四氢无生蛋白是一种酶的副因素及其在先天性缺乏障碍中补充可改善运动障碍。因此，可能有一个四氢无菌蛋白酶在运动障碍的发展中的关键作用，例如CP。我们开发了一个敲除兔，该兔在一种四氢无菌合成酶中引入特定突变，锯齿蛋白还原酶。胎儿缺氧 - 异常，新生儿兔子出现了高血压和困难保持平衡。离散大脑区域中表现出低发育四氢可毒素的胎儿兔子具有发展高渗。磁共振成像（MRI）使我们能够预测胎儿将发展产后高血压。这项进步允许识别早期关键途径引起高渗。我们的目标是阐明人类围产期脑损伤的分子机制突变引起了童年运动障碍，通过使用异性和兔子中的纯合敲除转基因方法。该提议中问的主要问题是是否选择性大脑区域中的四氢无菌蛋白毒素有助于患有严重性的运动障碍由增加产前侮辱（例如缺氧 - 缺血或炎症）确定。使用遗传基因敲除 Sepiapterin还原酶，我们可以进一步降低大脑中的四氢无菌水平，并研究是否是否产生的运动缺陷增加或我们需要较少的侮辱性才能达到相同的运动缺陷。第一个目的决定了增加的胎儿侮辱，缺氧 - 缺血或炎症是脂多糖，增强坟墓/纯合还原酶基因敲除中的运动障碍兔子。第二个目标将确定神经元或寡头损伤是否解释了淘汰兔子中的运动障碍。我们使用创新的产前和产后MRI生物标志物与组织流式细胞术和高性能液相色谱法的高血压与电化学检测。通过进行时间依赖性，器官特异性和细胞特异性的病原体研究，我们将获得该辅助因子在运动障碍的围产期发病机理中的作用的全面图片。