ATP Binding Cassette Transporters in Health and Disease

健康和疾病中的 ATP 结合盒转运蛋白

基本信息

批准号：
10237095
负责人：
Elizabeth Joanna Tarling
金额：
$ 60.81万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-09 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10237095
关键词：
ATP-Binding Cassette Transporters Acute Address Adult Alcohol consumption Animals Bile Acids Bile fluid Biliary Cannulations Cause of Death Cholestasis Classification Clustered Regularly Interspaced Short Palindromic Repeats Defect Development Disease Disease Progression Embryo Enzymes Event Fibrosis Functional disorder Gene Expression Genes Health Health system Hepatic Hepatomegaly Histologic Histopathology Human Hybrids Impairment Inbred Strains Mice Lead Lipids Liver Liver diseases Measures Metabolic Metabolism Methods Molecular Mus Nutrient Onset of illness Pathogenesis Pathogenicity Pathology Pathway interactions Physiological Processes Plasma Progressive intrahepatic cholestasis Proteins Proteomics Public Health Regulation Reporting Resistance Role Systems Biology Testing Therapeutic Intervention Triglycerides Very Long Chain Fatty Acid Virus Diseases Wild Type Mouse bile acid metabolism branched chain fatty acid chronic liver disease design disorder prevention effective therapy in vitro Assay in vivo lipid metabolism lipid transport lipidome lipidomics liver injury loss of function non-alcoholic fatty liver disease nonalcoholic steatohepatitis novel peroxisome prevent protein expression tool western diet

项目摘要

ABSTRACT Metabolic and chronic liver diseases are among the leading causes of death in the US. The liver is a central hub that coordinately regulates the metabolism of many nutrients, including lipids. The liver does not store lipids in the long-term, and lipid accumulation in the liver results in different diseases. Triglyceride accumulation in the liver causes steatosis which can progress to non-alcoholic steatohepatitis (NASH), both part of the non-alcoholic fatty liver disease (NAFLD) spectrum. Accumulation of bile acids in the liver because of viral infections, alcohol use or more advanced liver damage causes cholestasis. Identification of the molecular mechanisms of specific disease-promoting pathways is an essential step before pathways can be safely targeted for disease prevention. Our studies will further the understanding of the role of peroxisomal ABCD transporters in the liver. We have used an unbiased systems biology approach to identify new players in the regulation of lipid metabolism in the liver. Through these methods, we identified the peroxisomal transporter ABCD3 as a novel regulator of hepatic lipid metabolism. Abcd3−/− mice are partially lethal and loss of ABCD3 in surviving animals alters the hepatic lipidome and results in hepatomegaly and profoundly reduced biliary bile acids. To study ABCD3 in vivo, we have developed and validated a novel AAV-CRISPR strategy to disrupt Abcd3 exclusively in the liver, allowing us to disrupt Abcd3 in adult wild-type mice in a temporal fashion to determine the sequelae of events leading to the defects observed after loss of ABCD3. Using these tools, we show that acute loss of hepatic ABCD3 in adult mice is sufficient to recapitulate the dramatic reduction in biliary bile acids. When fed a Western diet (WD), loss of hepatic ABCD3 results in liver lipid accumulation as well as elevated plasma liver enzymes and bile acids, all hallmarks of NASH. We have designed two specific aims; in Aim 1 we will test the hypothesis that ABCD3 deficiency results in cholestasis and NASH. In Aim 2, we will identify specific substrates for ABCD3 and test the hypothesis that peroxisomal lipid defects are pathogenic and key for the development of NASH. Our studies demonstrate that loss of ABCD3, which is lethal in humans, results in cholestasis and NASH in a setting of elevated lipid levels. Completion of these studies will further the understanding of the role of peroxisomal ABCD transporters in the liver, and implicate peroxisomal lipid metabolism as an important contributor in the pathogenesis of NASH.

抽象的代谢和慢性肝脏疾病是美国死亡的主要原因之一。肝脏是中央集线器协调调节包括脂质在内的许多营养素的代谢。肝脏不会将脂质储存在肝脏中的长期和脂质积累会导致不同的疾病。甘油三酸酯在肝脏会导致脂肪变性，可以发展为非酒精性脂肪性肝炎（NASH），这两者都是非酒精性的脂肪肝病（NAFLD）谱系。由于病毒感染，酒精，胆汁酸积聚使用或更晚期的肝损害会导致胆汁淤积。鉴定特异性的分子机制促进疾病的途径是将途径安全针对预防疾病的必要步骤。我们的研究将进一步理解过氧化物体ABCD转运蛋白在肝脏中的作用。我们有使用公正的系统生物学方法来识别在调节脂质代谢中的新参与者肝。通过这些方法，我们确定了过氧化物体转运蛋白ABCD3是肝的新调节剂脂质代谢。 ABCD3 - / - 小鼠部分致命，幸存动物的ABCD3丧失改变了肝脂肪组并导致肝肿大，并严重降低了胆汁胆酸。为了在体内研究ABCD3，我们已经开发并验证了一种新颖的AAV-CRISPR策略，以专门破坏肝脏中的ABCD3，从而允许我们以临时方式破坏成年野生型小鼠的ABCD3，以确定事件的后遗症 ABCD3丢失后观察到的缺陷。使用这些工具，我们表明成人肝ABCD3的急性损失小鼠足以概括胆汁胆酸的急剧减少。喂西方饮食（WD）时，损失肝aBCD3的大量导致肝脂质积累以及血浆肝酶和胆汁酸的升高纳什的标志。我们设计了两个具体的目标。在AIM 1中，我们将测试ABCD3的假设缺乏会导致胆汁淤积和纳什。在AIM 2中，我们将确定ABCD3的特定基材并测试假设过氧化物体脂质缺陷是致病性的，并且是NASH发展的关键。我们的研究证明在人类中致命的ABCD3的损失导致胆汁淤积和纳什脂质水平升高。这些研究的完成将进一步理解过氧化物体ABCD的作用肝脏中的转运蛋白，并将过氧化物体脂质代谢牵涉到作为重要因素纳什的发病机理。