Methods to enable cholesterol catabolism in human monocyte derived macrophages

在人单核细胞衍生的巨噬细胞中实现胆固醇分解代谢的方法

• 批准号: 8337404
• 负责人: RICHARD E HONKANEN
• 金额: $ 33.41万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-23 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8337404
• 关键词: Address; Animal Model; Arterial Fatty Streak; Atherosclerosis; Bacteria; Bacterial Genes; Biochemical; Carbon Dioxide; Cardiovascular Diseases; Catabolism; Cause of Death; Cell Therapy; Cholestanes; Cholesterol; Cholesterol Esters; Clinical Trials; Complex; Coronary; Development; Disease; Enzymes; Event; Gene Expression; Genes; Genetic; Genus Mycobacterium; Goals; High Density Lipoproteins; Human; Immune response; Individual; Lead; Life Style; Lipoproteins; Medical; Methods; Myocardial Infarction; Pathway interactions; Phagosomes; Plasma; Production; Progressive Disease; Staging; Stroke; System; Testing; Transfection; United States; Vascular Diseases; age related; design; expression vector; feeding; high risk; innovation; lipid metabolism; macrophage; monocyte; novel strategies; prevent; promoter; response

DESCRIPTION (provided by applicant): Atherosclerotic cardiovascular disease (CVD) is the leading cause of death in the United States. CVD originates from aberrations in normal lipid metabolism (some genetic, some lifestyle choices) that result in elevated plasma lipoproteins (principally LDLs) and/or low levels of high-density lipoproteins (HDLs). For many people, CVD is an age dependent, progressive disease that is largely undetected or ignored until an event (i.e. myocardial infarction or stroke) occurs in the later stages of disease. Therefore, current therapies focus on preventing a second event (or a primary event in high risk individuals) by reducing the circulating levels of LDLs and/or increasing HDLs. However, at a biochemical level the inability of macrophages to degrade the cholestane ring of cholesterol is a fundamental component of CVD. If macrophages had the ability to degrade cholesterol, they would not become engorged with cholesterol/cholesterol esters and elicit the maladaptive immune response that leads to the onset and progression of atherosclerosis. Recently, studies of Mycobacteria survival in human macrophages revealed a surprising observation. Mycobacteria feed on cholesterol while contained in the phagosomes of macrophages. Importantly, two enzymes that catalyze cholestane ring opening have been identified. We plan to test the hypothesis that genes encoding enzymes identified in bacteria can be humanized and used to transformation human monocyte derived macrophages, enabling the degradation of phagosome-cholesterol. The main objectives are to: 1) humanize bacterial genes encoding key ring opening enzymes, 2) develop an innovative expressions systems to regulate the expression of these genes in response to changes in cellular levels of cholesterol, and 3) characterize the production and fate of compounds generated following B-ring opening. If this paradigm-challenging hypothesis is true, the proposed studies should lead to the development of an entirely new approach for the medical management of CVD.

描述（由申请人提供）：动脉粥样硬化心血管疾病（CVD）是美国死亡的主要原因。 CVD起源于正常脂质代谢（某些遗传，某些生活方式选择）的畸变，导致血浆脂蛋白升高（主要是LDL）和/或低水平的高密度脂蛋白（HDLS）。对于许多人来说，CVD是一种依赖年龄的，进行性疾病，直到在疾病后期发生事件（即心肌梗塞或中风）之前，它在很大程度上未被发现或忽略。因此，当前的疗法专注于通过降低LDL和/或增加HDL的循环水平来防止第二次事件（或高风险个体中的主要事件）。然而，在生化水平上，巨噬细胞降解胆固醇环是CVD的基本成分。如果巨噬细胞具有降解胆固醇的能力，它们将不会散发出胆固醇/胆固醇酯的能力，并引起适应不良的免疫反应，从而导致动脉粥样硬化的发作和进展。最近，对人类巨噬细胞中的分枝杆菌存活率的研究表明，观察到了令人惊讶的观察结果。分枝杆菌以胆固醇为食，同时包含在巨噬细胞的吞噬体中。重要的是，已经确定了两种催化cholestane环开口的酶。我们计划检验以下假设：在细菌中鉴定出的酶的基因可以人性化并用于转化人类单核细胞衍生的巨噬细胞，从而使吞噬体 - 胆固醇的降解。主要目标是：1）编码键环开放酶的细菌基因，2）开发一种创新的表达系统，以调节这些基因的表达，以响应胆固醇的细胞水平的变化，而3）表征B圈打开后产生的化合物的产生和命运。如果这种范式挑战的假设是正确的，那么拟议的研究应导致开发一种全新的CVD医学管理方法。