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: 8803354
• 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/8803354
• 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; Health; Heart; Heart failure; Herbicides; Human; Injury; Kidney; Lecithin; Left; Light; Light-Chain Immunoglobulins; Link; Liver; Malignant Neoplasms; Metabolism; Methods; Microvascular Dysfunction; 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; endothelial dysfunction; improved; in vivo; mortality; nanoliposome; nanoparticle; novel; novel strategies; prevent; primary amyloidosis of light chain type; protective effect; protein misfolding; 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，例如阿尔茨海默病（AD）和轻链淀粉样变性（AL）仍然是具有高发病率和死亡率的棘手疾病。 AL 是一种浆细胞恶性肿瘤，由免疫球蛋白轻链蛋白 (LC) 克隆过量产生，错误折叠并沉积在心脏、血管和其他组织中，如果不及时治疗，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接触橙剂有关，并指出其与退伍军人群体的特殊相关性。化疗——干细胞移植是唯一可用的治疗方法，具有较高的治疗相关死亡率和不完全成功率。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。淀粉样蛋白疾病（AL 和 AD）具有共同的病理生理学，涉及氧化应激和细胞凋亡损伤，这些损伤发生在暴露于可溶性前原纤维淀粉样蛋白的微血管和组织中（现已被认为是损伤的主要途径）。纳米脂质体（NLs）是直径<100 nm的人造磷脂囊泡，在治疗淀粉样蛋白疾病方面具有巨大潜力。与其他纳米粒子不同，纳米粒子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，减少细胞与淀粉样蛋白的接触。它们的表面可以进行化学修饰，以允许治疗货物的组织靶向和细胞内甚至细胞器特异性递送。在许多可以与 NL 缀合的治疗药物中，簇蛋白（也称为载脂蛋白 J）在防止淀粉样蛋白细胞损伤方面具有广阔的潜力。 Clusterin是一种组成型表达的伴侣糖蛋白，具有运输异常蛋白的主要双重作用，也是一种重要的促生存蛋白。 AL和AD都有代谢异常。我们的总体目标是通过开发和测试单独的纳米脂质体（未缀合）或与簇蛋白功能化（缀合）作为针对 LC 诱导的人体血管组织毒性的保护剂，来测试基于纳米脂质体的淀粉样蛋白疾病治疗。我们将测试以下假设：由胆固醇、鞘磷脂 (SM)、磷脂酰胆碱 (PC) 和磷脂酸 (PA) 组成的 NL 将与来自 AL 患者的 LC 结合，从而减少对离体人类脂肪小动脉的毒性。其次，我们将测试这样的假设：NLs 功能化以携带簇蛋白（通过非共价和共价方法）将允许簇蛋白在细胞周围和细胞内递送，这也将防止 LC 损伤。为了测试这些假设，在目标 1 中，我们将开发能够有效结合 LC 的 NL，并量化 NL 对 LC 引起的毒性的保护作用。在目标 2 中，我们将通过与凝聚素缀合来开发功能化的 NL，用于细胞内凝聚素递送，并测试缀合的 NL 对 LC 的保护作用 毒性。在这两个目标中，我们将探讨 NL 保护作用背后的关键机制。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，未来可能有助于治疗退伍军人和患有 AL 淀粉样变性以及其他淀粉样蛋白疾病（例如阿尔茨海默病）的患者。

项目成果

Oxidative Stress Alters the Morphology and Toxicity of Aortic Medial Amyloid.

氧化应激改变主动脉内侧淀粉样蛋白的形态和毒性。

• DOI: 10.1016/j.bpj.2015.10.034
• 发表时间: 2015-12-01
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Davies HA;Phelan MM;Wilkinson MC;Migrino RQ;Truran S;Franco DA;Liu LN;Longmore CJ;Madine J
• 通讯作者: Madine J

PEGylated-nanoliposomal clusterin for amyloidogenic light chain-induced endothelial dysfunction.

• DOI: 10.1080/08982104.2016.1274756
• 发表时间: 2018-06
• 期刊: Journal of liposome research
• 影响因子: 4.4
• 作者: Guzman-Villanueva D;Migrino RQ;Truran S;Karamanova N;Franco DA;Burciu C;Senapati S;Nedelkov D;Hari P;Weissig V
• 通讯作者: Weissig V