Transendothelial transport mediated modulations to the High Density Lipoprotein

跨内皮转运介导的高密度脂蛋白调节

• 批准号: 10245152
• 负责人: VASANTHY NARAYANASWAMI
• 金额: $ 11.06万
• 依托单位: CALIFORNIA STATE UNIVERSITY LONG BEACH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10245152
• 关键词: ATP-Binding Cassette Transporters; Address; Amphipathic Alpha Helix; Antiatherogenic; Antioxidants; Apolipoprotein A-I; Apolipoproteins; Arterial Fatty Streak; Arterial Intimas; Arylesterase; Attention; Biomedical Research; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cellular biology; Cholesterol; Collaborations; Communities; Development; Endothelial Cells; Evaluation; Exhibits; Fluorescent Probes; Funding; Future; Goals; High Density Lipoprotein Cholesterol; High Density Lipoproteins; Impairment; In Vitro; Joints; Knowledge; Leadership; Life Style; Light; Lipids; Lipoproteins; Literature; Location; Low Density Lipoprotein oxidation; Mass Spectrum Analysis; Mediating; Mentors; Mentorship; Metabolism; Modification; Molecular Conformation; Outcome; Particle Size; Patients; Pharmacology; Plasma; Post-Translational Protein Processing; Property; Proteins; Proteomics; Publications; Publishing; Reporting; Research; Research Training; Risk; Role; Structure; Structure-Activity Relationship; Students; Testing; Ursidae Family; antioxidant enzyme; apolipoprotein E-3; atheroprotective; base; biomarker identification; cardiovascular disorder risk; cardiovascular risk factor; conformational alteration; flexibility; gene therapy; improved; insight; lipidomics; macrophage; monolayer; particle; prevent; reconstitution; response; skills; symposium; training opportunity; transcytosis

Title: Transendothelial transport mediated modulations to the High Density Lipoprotein Summary Despite the vast amount of literature that indicates low plasma levels of HDL-cholesterol (HDL-C) is a risk factor for cardiovascular disease (CVD), the last decade has seen a paradigm shift in the concept that it may not be HDL-C levels per se but the functionality of HDL that is a determining factor in CVD. The shift in focus is in light of several studies that show that neither pharmacological nor genetic intervention to increase HDL-C levels, lower the risk for CVD. Thus, there is a pressing need to fill the gap in knowledge regarding the role of HDL in CVD from a mechanistic perspective and understand structure-function relationships in HDL. In the current proposal, we hypothesize that apolipoprotein (apo) AI and apoE3, two critical apos on HDL, undergo structural alterations and post translational modifications (PTM) and that HDL undergoes particle modulation as a consequence of transendothelial transport from plasma to the arterial intima with significant functional penalties. We will test this hypothesis by interrogating structure-function changes to the HDL with two specific aims: (1) Identify structural and proteomic alterations to the HDL associated with transendothelial transport across aortic endothelial cells, and, (2) Determine changes in HDL function following transendothelial transport. We will carry out spectroscopic analysis of reconstituted HDL (rHDL) bearing spatially sensitive fluorescent probes at flexible locations on apoAI or apoE3 to obtain insight into conformational alterations following transcytosis. Based on previous findings about oxidatively modified apoAI in atherosclerotic plaques, we postulate that the apos are susceptible to oxidative modification during transcytosis. To address this, we will determine PTM in transcytosed HDL, specifically on apoAI, apoE3 and selected proteins involved in lipoprotein metabolism by mass spectrometry. We will also perform lipoproteomic analysis to identify changes to the protein and lipid composition of transcytosed HDL. In an independent but complementary approach, we will examine changes in two major functional effects of transcytosed HDL: its ability to promote cholesterol efflux from macrophages, and, its antioxidant activity. Completion of these studies will significantly advance our understanding of the relationship between structure/composition and the atheroprotective effect of HDL at the vascular wall. The expected outcome of the proposed studies is to have a deeper understanding of modulations in HDL that render it dysfunctional. Establishing this relationship will aid in development of HDL- based therapies and identification of biomarkers of CVD risk. The research enhancement objectives of the PI are to: (i) develop expertise in HDL and endothelial cell biology, (ii) increase competitiveness to apply for major external funding by publishing research findings, (iii) strengthen existing and develop new collaborations, submit joint publications and lay the groundwork for developing proposals, and, (iv) establish mentoring, networking, and leadership skills, and offer research training opportunities and mentorship for students from diverse backgrounds in biomedical research.

标题：经内皮转运介导的高密度脂蛋白调节 总结 尽管大量的文献表明低血浆HDL-胆固醇（HDL-C）水平是一种风险， 心血管疾病（CVD）的因素，过去十年已经看到了概念的范式转变，它可能 不是HDL-C水平本身，而是HDL的功能，这是CVD的决定因素。焦点的转移是 鉴于几项研究表明，无论是药物还是遗传干预， 降低CVD的风险。因此，迫切需要填补有关知识方面的差距， 从机理的角度研究HDL在CVD中的作用，并了解HDL的结构-功能关系。 在目前的建议中，我们假设载脂蛋白（apo）AI和apoE 3，两个关键的HDL， 经历结构改变和翻译后修饰（PTM），HDL经历颗粒 作为从血浆到动脉内膜的跨内皮转运的结果， 功能性处罚。我们将通过询问HDL的结构-功能变化来验证这一假设， 两个具体的目标：（1）确定与跨内皮细胞相关的HDL的结构和蛋白质组学改变， 跨主动脉内皮细胞的转运，和（2）确定跨内皮细胞后HDL功能的变化 运输我们将进行光谱分析的重组HDL（rHDL）轴承空间敏感 在apoAI或apoE 3上的柔性位置处的荧光探针，以获得对构象改变的洞察 转胞吞作用后。基于先前关于动脉粥样硬化斑块中氧化修饰的apoAI的发现， 我们假设在转胞吞作用期间，pO对氧化修饰敏感。为了解决这个问题，我们 将确定转胞吞的HDL中的PTM，特别是apoAI、apoE 3和参与的选定蛋白质 脂蛋白代谢的质谱分析。我们还将进行脂蛋白组学分析，以确定变化 转胞吞HDL的蛋白质和脂质组成。在独立但互补的方法中，我们 我将研究转胞质HDL的两个主要功能效应的变化：它促进胆固醇的能力， 从巨噬细胞流出，以及其抗氧化活性。这些研究的完成将大大促进我们的 了解HDL的结构/组成与动脉粥样硬化保护作用之间的关系， 血管壁拟议研究的预期成果是更深入地了解 导致HDL功能失调建立这种关系将有助于HDL的发展- 基础治疗和CVD风险生物标志物的鉴定。 PI的研究增强目标是：（i）发展HDL和内皮细胞的专业知识 生物学，（ii）通过发表研究成果来提高申请主要外部资金的竞争力，（iii） 加强现有的合作并发展新的合作，提交联合出版物，并为 制定提案，以及（iv）建立指导、网络和领导技能，并提供研究 为来自不同背景的学生提供生物医学研究方面的培训机会和指导。

项目成果

Apolipoprotein E LDL receptor-binding domain-containing high-density lipoprotein: a nanovehicle to transport curcumin, an antioxidant and anti-amyloid bioflavonoid.

• DOI: 10.1016/j.bbamem.2010.09.007
• 发表时间: 2011-01
• 期刊: Biochimica et biophysica acta
• 作者: Khumsupan P;Ramirez R;Khumsupan D;Narayanaswami V
• 通讯作者: Narayanaswami V

Targeted intracellular delivery of resveratrol to glioblastoma cells using apolipoprotein E-containing reconstituted HDL as a nanovehicle.

• DOI: 10.1371/journal.pone.0135130
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Kim SH;Adhikari BB;Cruz S;Schramm MP;Vinson JA;Narayanaswami V
• 通讯作者: Narayanaswami V

Biochemical and biophysical characterization of recombinant rat apolipoprotein E: similarities to human apolipoprotein E3.

重组大鼠载脂蛋白 E 的生化和生物物理特征：与人载脂蛋白 E3 的相似性。

• DOI: 10.1016/j.abb.2012.10.007
• 发表时间: 2013
• 期刊: Archives of biochemistry and biophysics
• 影响因子: 3.9
• 作者: Tran,TuyenN;Kim,SeaH;Gallo,Carlos;Amaya,Max;Kyees,Jessica;Narayanaswami,Vasanthy
• 通讯作者: Narayanaswami,Vasanthy

Ordered opening of LDL receptor binding domain of human apolipoprotein E3 revealed by hydrogen/deuterium exchange mass spectrometry and fluorescence spectroscopy.

氢/氘交换质谱和荧光光谱显示人载脂蛋白 E3 的 LDL 受体结合域有序开放。

• DOI: 10.1016/j.bbapap.2018.08.005
• 发表时间: 2018
• 期刊: Biochimica et biophysica acta. Proteins and proteomics
• 作者: Yang,Liping;Hernandez,RoyV;Tran,TuyenN;Nirudodhi,Sasidhar;Beck,WendyHJ;Maier,ClaudiaS;Narayanaswami,Vasanthy
• 通讯作者: Narayanaswami,Vasanthy

Apolipoprotein E3-mediated cellular uptake of reconstituted high-density lipoprotein bearing core 3, 10, or 17 nm hydrophobic gold nanoparticles.

• DOI: 10.2147/ijn.s145326
• 发表时间: 2017
• 期刊: International journal of nanomedicine
• 影响因子: 8
• 作者: Chuang ST;Shon YS;Narayanaswami V
• 通讯作者: Narayanaswami V