Neprilysin and Peripheral Clearance of Amyloid Peptides

脑啡肽酶和淀粉样肽的外周清除

• 批准号: 7858439
• 负责人: Louis B. Hersh
• 金额: $ 18.48万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-15 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7858439
• 关键词: Abeta clearance; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Antibodies; Binding; Blood; Blood Cells; Blood Platelets; Bone Marrow Cells; Bone Marrow Stem Cell; Bone Marrow Transplantation; Brain; Cell membrane; Cell surface; Cells; Chimera organism; Chimeric Proteins; Complement; Enzymes; Epitopes; Erythrocytes; Ganglioside GM1; Gelsolin; Hematopoietic; Human; Hydrolysis; Impaired cognition; Injection of therapeutic agent; Leukocytes; Link; Lymphocyte; Measures; Methods; Mus; Neprilysin; Peptide Hydrolases; Peptides; Peripheral; Plasma; Play; Principal Investigator; Proteins; Public Health; Role; Subfamily lentivirinae; Tail; Testing; Therapeutic; Transgenic Mice; Translations; Transplantation; Veins; amyloid peptide; human subject; immunogenicity; morris water maze; mouse model; novel therapeutic intervention; prevent; programs

DESCRIPTION (provided by applicant): It has been shown that peripheral clearance of amyloid beta peptides reduces amyloid beta peptide levels in the brain through what has been termed a "sink effect". We have exploited this "sink effect" by showing that the peptidase neprilysin, when expressed peripherally on leukocytes reduced amyloid beta peptide levels in the brain of a hAPP transgenic mouse model of Alzheimer's disease. To obtain peripheral neprilysin expression we transplanted bone marrow stem cells transduced with lentivirus expressing neprilysin to a recipient hAPP transgenic mouse. We now propose to further develop peripheral expression of neprilysin as a therapeutic treatment for AD. Two specific aims directed at further developing peripheral neprilysin expression, without the need for bone marrow transplantation, are proposed: SPECIFIC AIM 1. To test the use of a single chain antibody-neprilysin (NEP) chimeric protein targeted to red blood cells to lower brain amyloid beta peptide levels. Chimeric proteins composed of a single chain antibody targeted to a mouse erythrocyte epitope fused with neprilysin will be used to produce erythrocyte bound neprilysin which will be tested for the ability to lower brain amyloid beta peptide levels. SPECIFIC AIM 2. To test the use of a myristoylated-neprilysin to attach NEP to hematopoietic cells. We will generate a form of neprilysin in which a myristoyl-peptide is linked to the C-terminus. This myristoyl-NEP will be used to produce peripheral NEP by insertion into the plasma membrane of hematopoietic cells and tested for the ability to lower brain amyloid beta peptide levels. We have as an overall objective the translation of one of these methods to the testing of human subjects. PUBLIC HEALTH RELEVENCE: The focus of this proposal is to develope a new therapeutic approach for treating Alzheimer's disease. We propose to express the enzyme neprilysin on blood cells where it can degrade plasma amyloid beta peptides leading to a lowering of brain amyloid beta peptides which are known to play a major role in causing Alzheimer's disease.

描述（由申请人提供）：已经表明，淀粉样蛋白β肽的外围清除通过称为“下水道效应”的淀粉样蛋白β肽可降低大脑中的淀粉样β肽水平。我们已经通过表明肽酶Neprilysin在白细胞上表达的肽酶Neprilysin降低了阿尔茨海默氏病的HAPP转基因小鼠模型的大脑中的淀粉样蛋白β肽水平，从而利用了这种“下沉效应”。为了获得周围的Neprilysin表达，我们将用慢跑病毒表达Neprilysin转导至受体HAPP转基因小鼠的慢病毒转导的骨髓干细胞。现在，我们建议进一步发展为Neprilysin作为AD治疗治疗的外周表达。提出了两个针对进一步发展的无需骨髓移植的外周酯表达的特定目的：具体目的1。测试使用单链抗体 - 涅prilysin（NEP）嵌合蛋白的使用，靶向降低脑淀粉蛋白蛋白质蛋白质降低大脑淀粉样蛋白蛋白质。由靶向小鼠红细胞表位的单链抗体组成的嵌合蛋白将用于产生红细胞结合的Neprilysin，该蛋白将测试，以降低脑淀粉样蛋白β肽水平。具体目的2。测试肉豆蔻酰化的静脉脂蛋白将NEP连接到造血细胞上。我们将生成一种形式的Neprilysin，其中肉豆蔻酰肽与C末端相关。该肉豆蔻酰NEP将用于通过插入造血细胞的质膜来产生周围NEP，并测试能够降低脑淀粉样蛋白β肽水平的能力。我们拥有总体目标，将其中一种方法转化为对人类受试者的测试。公共卫生相关性：该提案的重点是开发一种治疗阿尔茨海默氏病的新治疗方法。我们建议在血细胞上表达酶Neprilysin，它可以降解血浆淀粉样蛋白β肽，从而降低脑淀粉样蛋白β肽的降低，这些肽在引起阿尔茨海默氏病方面起着主要作用。