The role of Stearate in the regulation of vascular calcification in chronic kidne

硬脂酸盐在调节慢性肾血管钙化中的作用

基本信息

批准号：
8727657
负责人：
Makoto Miyazaki
金额：
$ 37.98万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8727657
关键词：
Acids Address Alkaline Phosphatase Apoptosis Arteries Attenuated Blood Vessels Ca(2+)-Transporting ATPase Calcium Cardiovascular Diseases Cardiovascular Physiology Cause of Death Cell Culture Techniques Chronic Chronic Kidney Failure Cyclic AMP-Dependent Protein Kinases Dietary Fats Echocardiography Endoplasmic Reticulum Enzymes Eukaryotic Initiation Factor-2 Event Fatty Acids Fluorescent Antibody Technique Goals Human In Vitro Knockout Mice Lead Lipids Mammalian Cell Measurement Mediating Metabolism Minerals Modeling Molecular Morphology Mus Osteoblasts Osteoclasts Osteogenesis Pathogenesis Pathway interactions Patients Phosphatidic Acid Phosphorylation Phosphotransferases Plant Roots Prevention strategy Process Protein Isoforms Proteins Regulation Reporting Research Role Saturated Fatty Acids Scheme Serum Smooth Muscle Myocytes Staining method Stains Stearates Stearic Acids Stearoyl-CoA Desaturase Structure Subfamily lentivirinae Techniques Testing Translations Vascular calcification Western Blotting Wild Type Mouse absorption calcification design effective therapy endoplasmic reticulum stress fluidity human MYH11 protein in vitro Model in vivo inhibitor/antagonist lipid biosynthesis lipine loss of function mineralization mouse model osteogenic overexpression protein expression public health relevance research study response small hairpin RNA transcription factor transcription factor CHOP

项目摘要

DESCRIPTION (provided by applicant): Cardiovascular disease such as vascular calcification is a leading cause of death in subjects with CKD. However, there is no effective therapy for vascular calcification available. Stearate, a major saturated fatty acid in mammalian cells, is derived through two pathways: dietary fat absorption and de novo lipogenesis. Our long-term goal is to identify new pharmacological strategies for the prevention of vascular calcification. Our previous study has revealed that stearate derived from de novo lipogenesis promotes aortic calcification. Both stearoyl-CoA desaturase (SCD)-1 and SCD2 are central enzymes involved in the metabolism of stearic acid. As shown in our preliminary results, increasing stearate by adding exogenous stearate or by inhibiting SCD1 and SCD2 augmented vascular calcification in vitro and in vivo. Recently, we found that levels of serum stearate were significantly increased in subjects with chronic kidney disease (CKD), due to lower SCD activity. The pro-calcific and pro-osteogenic effects of stearate were highly correlated with levels of distearoyl-phosphatidic acid (18:0-PA) in the ER and the expression of ATF4, which is a central transcription factor in the regulation of osteogenesis. These effects were also associated with the expression of CHOP, a transcription factor that promotes apoptosis. We therefore hypothesize that 18:0-PA derived from stearate synthesized by SCD in the endoplasmic reticulum induces vascular osteogenesis and calcification by activating the PERK-eIF2¿-ATF4-CHOP pathway of the unfolded protein response in vivo. We will design experiments to address this hypothesis using in vivo and in vitro approaches. The two specific aims of this proposal are 1) To determine the molecular mechanism underlying stearate-mediated vascular calcification in vitro, and 2) To determine whether the activation of the ATF4-CHOP pathway mediates vascular calcification induced by increased stearate due to VSMC deficiency of SCD1 and SCD2 in vivo.

描述（由申请人提供）：心血管疾病（例如血管钙化）是 CKD 患者死亡的主要原因。然而，硬脂酸是哺乳动物细胞中的一种主要饱和脂肪酸，目前还没有有效的治疗方法。两种途径：膳食脂肪吸收和从头脂肪生成我们的长期目标是确定预防血管钙化的新药理学策略。 novo 脂肪生成促进主动脉钙化。硬脂酰辅酶 A 去饱和酶 (SCD)-1 和 SCD2 都是参与硬脂酸代谢的核心酶，通过添加外源性硬脂酸或抑制 SCD1 和 SCD2 可以增加硬脂酸含量。最近，我们发现血清硬脂酸盐水平显着升高。患有高度慢性肾病 (CKD) 的受试者，由于 SCD 活性较低，硬脂酸盐的促钙化和促骨生成作用与 ER 中二硬脂酰磷脂酸 (18:0-PA) 的水平以及ATF4 是骨生成调节的核心转录因子，这些效应也与促进细胞凋亡的转录因子 CHOP 的表达有关。因此，我们捕获了源自 18:0-PA 的信号。内质网中 SCD 合成的硬脂酸盐通过激活 PERK-eIF2 诱导血管成骨和钙化体内未折叠蛋白反应的 ATF4-CHOP 途径我们将使用体内和体外方法设计实验来解决这一假设，该提案的两个具体目标是 1) 确定硬脂酸介导的血管钙化的分子机制。 2) 确定 ATF4-CHOP 通路的激活是否介导由于体内 VSMC 缺乏 SCD1 和 SCD2 导致硬脂酸盐增加诱导的血管钙化。