Nanoliposome-based Treatment of Amyloid Protein (AL) Toxicity

基于纳米脂质体的淀粉样蛋白 (AL) 毒性治疗

• 批准号: 8543427
• 负责人: Raymond Quezon Migrino
• 金额: --
• 依托单位: PHOENIX VA HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8543427
• 关键词: Address; Adipose tissue; Affinity; Alzheimer' s Disease; Amino Acids; Amyloid; Amyloid Proteins; Amyloidosis; Apoptosis; Apoptotic; Binding; Blood Vessels; Caliber; Cardiolipins; Cells; Charge; Cholesterol; Coupling; Deposition; Development; Disease; Electrostatics; Exposure to; Functional disorder; Future; General Population; Glycoproteins; Goals; Heart; Heart failure; Herbicides; Human; Injury; Kidney; Lecithin; Left; Light; Light-Chain Immunoglobulins; Link; Liver; Malignant Neoplasms; Metabolism; Methods; Molecular Chaperones; Morbidity - disease rate; Organ; Organelles; Oxidative Stress; Parkinson Disease; Patients; Phosphatidic Acid; Phospholipids; Plasma Cell Neoplasm; Plasma Cells; Population; Production; Protective Agents; Proteins; Protocols documentation; Recombinants; Reporting; Research; Role; Smooth Muscle Myocytes; Sphingomyelins; Stem cell transplant; Surface; Technology; Testing; Therapeutic; Tissues; Toxic effect; Variant; Vesicle; Veterans; agent orange; arteriole; base; cell injury; chemotherapy; improved; in vivo; mortality; nanoliposome; nanoparticle; novel; novel strategies; prevent; primary amyloidosis of light chain type; protective effect; protein misfolding; public health relevance; response; success; sulfated glycoprotein 2; tool

DESCRIPTION (provided by applicant): Amyloid protein-misfolding diseases such as Alzheimer's (AD) and light chain amyloidosis (AL) remain intractable diseases with high morbidity and mortality. In AL, a plasma cell malignancy from clonal overproduction of immunoglobulin light chain proteins (LC) that misfold, and deposit in the heart, vessels and other tissues, median survival is 4 months if left untreated. AL is a malignancy that the VA recognizes as being related to Agent Orange exposure, pointing to its specific relevance in the veteran population. Chemotherapy¿stem cell transplantation, the only treatment available, has high treatment- related mortality and incomplete success rate. We currently lack options to directly treat or protect cells against amyloid protein injury. Amyloid diseases (AL and AD) share common pathophysiology involving oxidative stress and apoptotic injury that occur early in microvessels and tissues exposed to soluble prefibrillar amyloid proteins (now recognized as a major path to injury). Nanoliposomes (NLs) are artificial phospholipid vesicles with a diameter of <100 nm that have great potential for treating amyloid protein disease. Unlike other nanoparticles, NLs are ideal for human treatment because they are non- toxic, non-immunogenic, fully biodegradable and structurally versatile. They can bind amyloid proteins with high affinity in vivo, reducing cell exposure to amyloid proteins. Their surface can be chemically modified to allow tissue targeting and intracellular and even organelle-specific delivery of therapeutic cargo. Among many therapeutic cargos that can be conjugated to NLs, clusterin (also known as apolipoprotein J) has promising potential to protect against amyloid protein cell injury. Clusterin is a constitutively expressed chaperone glycoprotein that has major dual roles of transporting abnormal proteins and is also an important pro-survival protein. It has abnormal metabolism in both AL and AD. Our overall goal is to test nanoliposome-based treatment of amyloid protein disease by developing and testing nanoliposomes alone (unconjugated) or functionalized with clusterin (conjugated) as protective agents against LC-induced toxicity to human vascular tissue. We will test the hypothesis that NLs composed of cholesterol, sphingomyelin (SM), phosphatidylcholine (PC) and phosphatidic acid (PA) will bind LC derived from AL patients thereby reducing toxicity to ex-vivo human adipose arterioles. Second, we will test the hypothesis that NLs functionalized to carry clusterin (both through non-covalent and covalent methods) will allow pericellular and intracellular delivery of clusterin that will also protect against LC injury. To test these hypotheses, in aim 1 we will develop NLs that will efficiently bind LC and quantify NLs' protective effect against LC induced toxicity. In aim 2, we will develop functionalized NLs through conjugation with clusterin for use in intracellular clusterin delivery and test protective effect of conjugated NLs against LC toxicity. In both aims, we will probe key mechanisms underlying the protective effects of NLs. The proposal represents a novel approach to the treatment of amyloid protein disease using nanoparticles that might be useful for the treatment of veterans and patients with AL amyloidosis and other amyloid diseases such as Alzheimer's disease in the future.

描述(由申请人提供)： 淀粉样蛋白错误折叠疾病，如阿尔茨海默病(AD)和轻链淀粉样变性(AL)，仍是高发病率和高死亡率的顽固性疾病。AL是一种浆细胞恶性肿瘤，由于免疫球蛋白轻链蛋白(LC)的克隆性过度产生，错误折叠并沉积在心脏、血管和其他组织中，如果不治疗，中位生存期为4个月。AL是一种恶性肿瘤，退伍军人管理局认为与橙剂暴露有关，指出它在退伍军人群体中具有特定的相关性。化疗干细胞移植是唯一可用的治疗方法，但与治疗相关的死亡率和不完全成功率很高。我们目前缺乏直接治疗或保护细胞免受淀粉样蛋白损伤的选择。淀粉样变性疾病(AL和AD)具有共同的病理生理机制，涉及氧化应激和细胞凋亡损伤，这些损伤早期发生在暴露于可溶性纤维前淀粉样蛋白(现已被认为是损伤的主要途径)的微血管和组织中。纳米脂质体(NLS)是一种直径为100 nm的人工磷脂微囊，在治疗淀粉样蛋白疾病方面具有巨大的潜力。与其他纳米粒子不同，NLS是人类治疗的理想选择，因为它们无毒、无免疫原性、完全可生物降解且结构多样。它们可以在体内以高亲和力结合淀粉样蛋白，减少细胞对淀粉样蛋白的暴露。它们的表面可以进行化学修饰，以允许组织靶向和细胞内甚至细胞器特定的治疗货物输送。在许多可与NLS结合的治疗性药物中，Clusterin(也称为载脂蛋白J)具有保护淀粉样蛋白细胞损伤的前景。Clusterin是一种结构性表达的伴侣糖蛋白，具有运输异常蛋白的双重作用，也是一种重要的促生存蛋白。急性白血病和阿尔茨海默病均存在代谢异常。我们的总体目标是通过单独开发和测试纳米脂质体(未结合)或与聚集素功能化(结合)作为保护剂来对抗LC对人类血管组织的毒性，来测试基于纳米脂质体的淀粉样蛋白疾病的治疗。我们将验证由胆固醇、鞘磷脂(SM)、磷脂酰胆碱(PC)和磷脂酸(PA)组成的NLS将结合AL患者来源的LC从而降低对体外人类脂肪小动脉的毒性的假设。其次，我们将检验这样一种假设，即NLS被功能化以携带Clusterin(通过非共价和共价方法)将允许细胞外和细胞内传递Clusterin，这也将保护LC免受损伤。为了验证这些假设，在目标1中，我们将开发NLS，它将有效地结合LC，并量化NLS对LC毒性的保护作用。在目标2中，我们将通过与Clusterin的偶联来开发功能化的NLS，用于细胞内的Clusterin递送，并测试结合的NLS对LC的保护作用 毒性。在这两个目标中，我们将探索NLS保护作用的关键机制。该提案代表了一种使用纳米颗粒治疗淀粉样蛋白疾病的新方法，这种方法可能对未来退伍军人和AL淀粉样变性患者以及阿尔茨海默病等其他淀粉样疾病的治疗有用。