NPC1L1 and Metabolic Diseases

NPC1L1 与代谢疾病

• 批准号: 8239578
• 负责人: Liqing Yu
• 金额: $ 32.3万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8239578
• 关键词: ASBT protein; Address; Adipose tissue; Affect; Age; Animals; Bile Acids; Biliary; Biochemical; Biochemistry; Blood; Body Size; Body Weight; Brain; Brown Fat; Catabolism; Cholesterol; Cholesterol Homeostasis; Data; Development; Diet; Disease; Dyslipidemias; Eating; Energy Metabolism; Enterocytes; Enterohepatic Circulation; Enzymes; Etiology; Excretory function; FDA approved; Fasting; Fatty Liver; Fatty acid glycerol esters; Feces; Gatekeeping; Gender; Gene Expression; Genes; Genetic; Glucose; Goals; Heart; Homeostasis; Hormones; Human; Inflammation; Insulin Resistance; Intestines; Iodothyronine Deiodinase; Knock-out; Knockout Mice; Link; Lipids; Liver; Liver diseases; Measures; Messenger RNA; Metabolic; Metabolic Diseases; Metabolism; Molecular; Molecular Probes; Morbidity - disease rate; Mus; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Obesity; Organ; Pathway interactions; Phenotype; Physical activity; Pilot Projects; Prevention; Proteins; Public Health; Relative (related person); Reproduction; Research; Resistance; Risk Factors; Role; Signaling Molecule; Skeletal Muscle; Small Intestines; Societies; Testing; Thyroid Hormones; Tissues; Transgenic Mice; Weight Gain; Wild Type Mouse; base; cholesterol absorption; cholesterol biosynthesis; clinically relevant; cytokine; dietary excess; ezetimibe; feeding; gene function; global health; glucose metabolism; human tissue; ileum; inhibitor/antagonist; insight; insulin sensitivity; lipid metabolism; mortality; non-alcoholic fatty liver; novel; novel strategies; obesity treatment; prevent; public health relevance; uptake

DESCRIPTION (provided by applicant): Niemann-Pick C1-Like 1 (NPC1L1) was originally identified as an essential protein for intestinal cholesterol absorption. It is the target of ezetmibe, a cholesterol absorption inhibitor that is widely used to lower blood cholesterol in humans. Surprisingly, genetic inactivation or ezetimibe inhibition of NPC1L1 in mice was recently found to protect against high fat diet (HFD)-induced obesity (DIO) and associated metabolic disorders. The molecular mechanisms underlying these novel findings remain unknown. The overall goal of the proposed studies is to define how NPC1L1 deficiency affects DIO. In preliminary studies, we found that food intake was identical between wild-type (WT) and NPC1L1 knockout (L1KO) mice, yet L1KO mice were protected against DIO and fatty liver, which was associated with increased expression of genes promoting energy expenditure in brown adipose tissue (BAT) and skeletal muscle. Thus, we hypothesize that NPC1L1 deficiency protects mice from DIO by increasing energy expenditure. We will directly measure energy expenditure, and molecularly define how NPC1L1 deficiency stimulates energy dissipation by measuring metabolic changes at biochemical and gene expression levels in WT and L1KO mice under dietary excess, ezetimibe treatment or fasting conditions. In mice, NPC1L1 is highly expressed in small intestine and only low levels of NPC1L1 mRNA can be detected in non-intestinal tissues. We hypothesize that loss of intestinal NPC1L1 is responsible for resistance to DIO. To directly test this hypothesis, we will cross our newly-created transgenic mice specifically expressing human NPC1L1 in small intestine to L1KO mice, thereby re-establishing intestinal NPC1L1 expression, and determine if this will rescue weight gain in L1KO mice. We will also determine if NPC1L1 deficiency requires a thyroid hormone activating enzyme type 2 iodothyronine deiodinase (D2) to prevent DIO since we found that D2 expression was substantially elevated in BAT and muscle in L1KO mice, and D2 activation is known to promote energy expenditure. To probe the molecular link between NPC1L1 deficiency and D2 activation, we will examine if bile acid metabolism differs between WT and L1KO mice on HFD because we found that many bile acid sensitive genes were upregulated in the ileum of L1KO mice and bile acids can function as signaling molecules to stimulate energy expenditure via a D2- dependent mechanism. Taken together, these studies will provide important new insight into the role of NPC1L1 in metabolic diseases, and have the potential to reveal new approaches for the prevention and treatment of obesity and associated metabolic risk factors, which contribute substantially to disease morbidity and mortality. PUBLIC HEALTH RELEVANCE: The overall goal of this proposal is to define how genetic inactivation or pharmacologic inhibition of intestinal NPC1L1 influences the development of obesity induced by a high fat diet. Findings from proposed studies hold the promise of revealing new approaches for the prevention and treatment of metabolic diseases such as obesity, type 2 diabetes and nonalcoholic fatty liver diseases. Given that NPC1L1 is the target of ezetimibe, an FDA-approved intestinal cholesterol absorption inhibitor that is widely used to lower blood cholesterol in humans, these studies have enormous translational potential.

描述（由申请方提供）：尼曼-匹克C1样蛋白1（NPC 1 L1）最初被鉴定为肠道胆固醇吸收的必需蛋白。它是ezetmibe的靶点，ezetmibe是一种胆固醇吸收抑制剂，广泛用于降低人类的血液胆固醇。令人惊讶的是，最近发现小鼠中NPC 1 L1的基因失活或依折麦布抑制可防止高脂饮食（HFD）诱导的肥胖（DIO）和相关代谢紊乱。这些新发现背后的分子机制仍然未知。拟议研究的总体目标是确定NPC 1 L1缺陷如何影响DIO。在初步研究中，我们发现野生型（WT）和NPC 1 L1基因敲除（L1 KO）小鼠之间的食物摄入量相同，但L1 KO小鼠可防止DIO和脂肪肝，这与棕色脂肪组织（BAT）和骨骼肌中促进能量消耗的基因表达增加有关。因此，我们假设NPC 1 L1缺陷通过增加能量消耗来保护小鼠免受DIO。我们将直接测量能量消耗，并通过测量WT和L1 KO小鼠在饮食过量、依折麦布治疗或禁食条件下生化和基因表达水平的代谢变化，从分子水平上确定NPC 1 L1缺乏如何刺激能量耗散。在小鼠中，NPC 1 L1在小肠中高度表达，而在非肠组织中只能检测到低水平的NPC 1 L1 mRNA。我们推测，肠道NPC 1 L1的丢失是对DIO耐药的原因。为了直接验证这一假设，我们将新创建的在小肠中特异性表达人NPC 1 L1的转基因小鼠与L1 KO小鼠杂交，从而重建肠道NPC 1 L1表达，并确定这是否会挽救L1 KO小鼠的体重增加。我们还将确定NPC 1 L1缺乏是否需要甲状腺激素激活酶2型碘甲腺原氨酸脱碘酶（D2）来预防DIO，因为我们发现L1 KO小鼠的BAT和肌肉中D2表达显著升高，并且已知D2激活可促进能量消耗。为了探索NPC 1 L1缺陷和D2激活之间的分子联系，我们将检查HFD的WT和L1 KO小鼠之间的胆汁酸代谢是否不同，因为我们发现许多胆汁酸敏感基因在L1 KO小鼠的回肠中上调，胆汁酸可以作为信号分子通过D2依赖性机制刺激能量消耗。综上所述，这些研究将为NPC 1 L1在代谢性疾病中的作用提供重要的新见解，并有可能揭示预防和治疗肥胖及相关代谢风险因素的新方法，这些因素对疾病发病率和死亡率有很大影响。 公共卫生关系：本提案的总体目标是确定肠道NPC 1 L1的遗传失活或药理学抑制如何影响由高脂饮食诱导的肥胖症的发展。拟议研究的结果有望揭示预防和治疗代谢性疾病的新方法，如肥胖，2型糖尿病和非酒精性脂肪肝疾病。考虑到NPC 1 L1是依折麦布的靶点，依折麦布是一种FDA批准的肠道胆固醇吸收抑制剂，广泛用于降低人体血液胆固醇，这些研究具有巨大的转化潜力。