Development of a novel treatment for hyperbilirubinemia-induced kernicterus

开发治疗高胆红素血症引起的核黄疸的新疗法

基本信息

批准号：
9926721
负责人：
DARIA MOCHLY-ROSEN
金额：
$ 32.94万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9926721
关键词：
Address Adult Affect African American Age Aldehydes Animal Model Antioxidants Asians Auditory Basic Science Bilirubin Binding Sites Brain Injuries Cells Cessation of life Clinical Clinical Trials Complex Crystallization Crystallography Cultured Cells Developed Countries Developing Countries Development Drug Screening Enzymes Erythrocytes Family Genes Genetic Diseases Glucosephosphate Dehydrogenase Glucosephosphate Dehydrogenase Deficiency Glutathione Goals Gunn Rats Hemolysis High Prevalence Hospitals Human Hyperbilirubinemia Icterus Impairment In Vitro Incidence Infant Kernicterus Knock-in Letters Measures Modeling Molecular Molecular Chaperones Morbidity - disease rate Motor Mutation Myocardial Infarction NADP Neonatal Nervous System Trauma Neurologic Deficit Neuronal Injury Neurons Neurosciences Neurosciences Research Newborn Infant Parkinson Disease Pathologic Pharmaceutical Chemistry Pharmaceutical Preparations Phototherapy Point Mutation Population Positioning Attribute Predisposition Production Radiodermatitis Rattus Registries Research Risk Rodent Model Roentgen Rays Severities Societies Structural defect Structure Structure-Activity Relationship Testing Time Toxic effect Transgenic Mice Whole Blood Exchange Transfusion aldehyde dehydrogenases clinical care cytotoxicity disability drug development drug discovery epidemiology study experience human disease human model in vivo mortality motor disorder mouse model mutant neurotoxicity novel novel strategies novel therapeutics prevent protein structure public health relevance screening skills small molecule small molecule libraries

项目摘要

DESCRIPTION (provided by applicant): Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common genetic disorder in humans, with more than 160 point mutations in this gene, and over 400 million people affected. Although most research relates to G6PD deficiency and increased hemolysis (breakdown of red blood cells) and accumulation of bilirubin in adults, epidemiological studies indicate that G6PD deficiency is also a major cause of pathologic neonatal bilirubin accumulation (jaundice) and a contributor to morbidity, including neurological injury (kernicterus). The hypothesis to be tested here is that correcting the activity and stability of G6PD mutants as well as increasing the activity of the wild-type enzyme will decrease bilirubin-induced neurotoxicity in infants. In this proposal, the plan to begin developing a treatment for kernicterus using a totally novel approach, by developing activators of wild-type (Wt) and mutant G6PDs is described. The project includes four aims: AIM 1: Identify small molecule chaperones that increase the catalytic activity of Wt and common G6PD mutants using an in vitro screen of a library of small molecules. AIM 2: Determine the X-ray crystal structure of the Wt and mutant G6PD enzymes in complex with the G6PD chaperone(s). Aim 3: Evaluate the ability of the chaperone(s) to protect cultured cells expressing Wt or the mutant G6PDs from bilirubin-induced cytotoxicity and elucidate the molecular basis of their effects. AIM 4: Evaluate the ability of the small molecule chaperone(s) to reduce bilirubin-induced neurotoxicity in rodent models of newborn hyperbilirubinemia, in vivo. Two established rat models of kernicterus (Gunn rats and bilirubin-injected Wt rats) and three new mouse models [transgenic mice, mimicking three common G6PD mutations with kernicterus] will be used. Small molecules, identified in Aim 3 to protect neurons from bilirubin-induced toxicity, will be tested for their ability to prevent or reduce neurological injury (measured as loss of motor and auditory skills) due to hyperbilirubinemia in one or more of these rodent models, in vivo. The experience of Dr. Stevenson (co-PI) in basic research related to hyperbilirubinemia and in clinical care of such newborns, the expertise of Dr. Wakatsuki in crystallography, together with our expertise in drug discovery and development and in neuroscience research, and the expertise of other advisors in drug screening, medicinal chemistry, in gene editing and in assessing auditory and motor dysfunctions, place the team in a unique position to address the above goals.

描述（由申请人提供）：6-磷酸葡萄糖脱氢酶（G6PD）缺乏症是人类最常见的遗传性疾病，该基因有超过 160 个点突变，超过 4 亿人受到影响，尽管大多数研究都与 G6PD 缺乏症有关。以及成人溶血（红细胞分解）和胆红素积累增加，流行病学研究表明 G6PD 缺乏也是病理性新生儿的主要原因胆红素积累（黄疸）和发病率的一个因素，包括神经损伤（核黄疸）。这里要检验的假设是纠正活动。 G6PD 突变体的稳定性和稳定性以及增加野生型酶的活性将减少胆红素引起的婴儿神经毒性。描述了一种使用全新方法治疗核黄疸的方法，即开发野生型 (Wt) 和突变型 G6PD 激活剂。该项目包括四个目标：目的 1：识别可增加 Wt 和常见 G6PD 突变体催化活性的小分子伴侣。使用小分子文库的体外筛选目的 2：确定与 G6PD 复合的 Wt 和突变型 G6PD 酶的 X 射线晶体结构。目标 3：评估伴侣保护表达 Wt 或突变型 G6PD 的培养细胞免受胆红素诱导的细胞毒性的能力，并阐明其作用的分子基础。目标 4：评估小伴侣的能力。分子伴侣可减少新生高胆红素血症啮齿动物模型中胆红素诱导的神经毒性，体内两种已建立的大鼠模型。将使用核黄疸（Gunn 大鼠和注射胆红素的 Wt 大鼠）和三种新的小鼠模型（转基因小鼠，模拟三种常见的核黄疸 G6PD 突变），在目标 3 中确定可保护神经元免受胆红素诱导的毒性。在这些啮齿动物模型中的一种或多种体内测试了它们预防或减少由于高胆红素血症引起的神经损伤（以运动和听觉技能丧失为衡量标准）的能力。 Stevenson 博士（联合 PI）在与高胆红素血症相关的基础研究和此类新生儿的临床护理方面的经验，Wakatsuki 博士在晶体学方面的专业知识，以及我们在药物发现和开发以及神经科学研究方面的专业知识，以及药物筛选、药物化学、基因编辑以及评估听觉和运动功能障碍方面的其他顾问使该团队处于实现上述目标的独特地位。