Re-engineering Erythropoietin for Alzheimer's Disease

重新设计促红细胞生成素治疗阿尔茨海默病

基本信息

批准号：
9297041
负责人：
Rachita Sumbria
金额：
$ 19.54万
依托单位：
KECK GRADUATE INST OF APPLIED LIFE SCIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-15 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9297041
关键词：
Address Adverse effects Alzheimer&apos s Disease Alzheimer&apos s disease model Amyloid beta-Protein Attenuated Basic Science Binding Biological Blood - brain barrier anatomy Blood Circulation Brain Chemistry Chimeric Proteins Chronic Chronic Disease Clinical Research Clinical Trials Cognition Cognitive Cognitive deficits Deoxyuridine Development Disease Disease Progression Dose Drug Delivery Systems Engineering Enzyme-Linked Immunosorbent Assay Erythropoietin Functional disorder Goals Hematocrit procedure Hematopoietic Histologic Hypertension ITGAM gene Immune Impaired cognition Inflammation Mediating Molecular Weight Monoclonal Antibodies Mus Nerve Degeneration Nerve Regeneration Neurodegenerative Disorders Neurons Organ Oxidative Stress Pathogenesis Pathology Penetration Performance Peripheral Plasma Primates Process Property Research Rodent Route Safety Serum Synapses Synaptophysin TFRC gene Testing Therapeutic Thioflavin S Thrombosis Transferrin B Transgenic Mice abeta toxicity analog base cognitive function cytokine drug candidate drug development improved memory recognition neurogenesis neuroinflammation neuronal survival neurotransmission neurotrophic factor neurotropin novel object recognition prevent receptor recombinant human erythropoietin targeted agent therapeutic candidate

项目摘要

PROJECT SUMMARY / ABSTRACT The goal of the current proposal is to develop a brain penetrating erythropoietin (EPO) analogue with negligible hematopoietic side effects, for AD. EPO is a unique therapeutic candidate for AD since it targets a spectrum of process that are involved in the pathophysiology and disease progression of AD, and importantly, promotes neurogenesis and improves cognition. This is clearly distinct from the numerous anti-Aβ agents that are currently under development for AD since anti-Aβ agents do not reverse the existing neuronal damage or cognitive decline. EPO on the other hand has both neuroprotective and neuroregenerative properties. This provides the scientific rationale to develop EPO for AD. Two major obstacles to the development of EPO for AD are: a) limited blood-brain barrier (BBB) penetration necessitating transcranial administration or use of high doses, and b) high doses of EPO result in undesired hematopoietic effects. To achieve our goal, a fusion protein of EPO with a monoclonal antibody (MAb) against the mouse transferrin receptor (TfR) has been engineered and is designated as cTfRMAb-EPO. The cTfRMAb-EPO fusion protein offers dual advantages: a) it rapidly enters the brain via the BBB TfR and b) it is rapidly cleared from the systemic circulation via the peripheral TfR, resulting in negligible hematopoietic effects. Our central hypothesis is that chronic systemic administration of the cTfRMAb-EPO fusion protein modifies multiple targets of AD pathophysiology and disease progression (Aβ-pathology (↓), neuroinflammation (↓), synaptic loss (↓), neurogenesis (↑), cognitive function (↑), and is thus therapeutic in AD with negligible hematopoietic effects. In the proposed studies, APPswe, PSEN1dE9 (APP/PS1) mice will be treated with either the cTfRMAb-EPO fusion protein, recombinant human EPO (rhuEPO) or vehicle, and the following specific aims will be addressed: Aim 1: Determine the cytoprotective effects of the cTfRMAb-EPO fusion protein in the APP/PS1 mice. Specifically, microglial activation (CD11b), neuroinflammation (brain cytokine levels), neurodegeneration (FluoroJade C), synaptic loss (synaptophysin) and neurogenesis (5-bromo-2′-deoxyuridine immunostaining) will be studied. Additionally, we will also study the effect of the brain-penetrating EPO on hematocrit, serum chemistry and histological effects on major organs following chronic dosing; Aim 2: Characterize the effects of the cTfRMAb-EPO fusion protein on cognitive deficits in the APP/PS1 mice. In particular, recognition memory and exploration will be assessed using the novel object recognition and open-field activity tests; Aim 3: Determine the effects of the cTfRMAb-EPO fusion protein on Aβ-pathology in the APP/PS1 mice. Specifically, the effect of the cTfRMAb- EPO fusion protein on Aβ plaque burden will be studied. The proposal will underscore the need for a neuroprotective-neuroregenerative approach for AD and provide a drug-delivery strategy to deliver EPO to the brain with negligible hematopoietic side effects.

项目概要/摘要当前提案的目标是开发一种脑穿透性促红细胞生成素 (EPO) 类似物，其含量可以忽略不计。造血副作用，因为 EPO 是治疗 AD 的独特候选药物，因为它针对一系列疾病。参与 AD 的病理生理学和疾病进展的过程，重要的是，促进这与众多抗 Aβ 药物明显不同。目前正在开发针对 AD 的药物，因为抗 Aβ 药物不能逆转现有的神经损伤或另一方面，EPO 具有神经保护和神经再生特性。为AD开发EPO提供了科学依据。 AD 是： a) 血脑屏障 (BBB) 穿透有限，需要经颅给药或使用高浓度 b) 高剂量的 EPO 会导致不良的造血作用为了实现我们的目标，融合。 EPO 蛋白与抗小鼠转铁蛋白受体 (TfR) 的单克隆抗体 (MAb) 工程化并命名为 cTfRMAb-EPO 融合蛋白具有双重优势：a) 它通过 BBB TfR 快速进入大脑，并且 b) 通过外周 TfR，导致可忽略不计的造血作用。我们的中心假设是慢性全身性。 cTfRMAb-EPO 融合蛋白的施用可改变 AD 病理生理学的多个靶点疾病进展（Aβ-病理 (↓)、神经炎症 (↓)、突触丧失 (↓)、神经发生 (↑)、认知功能 (↑)，因此对 AD 具有治疗作用，但造血作用可以忽略不计。 APPswe、PSEN1dE9 (APP/PS1) 小鼠将用 cTfRMAb-EPO 融合蛋白、重组蛋白进行治疗人类 EPO (rhuEPO) 或车辆，并将解决以下具体目标：目标 1：确定 cTfRMAb-EPO 融合蛋白对 APP/PS1 小鼠的细胞保护作用，特别是小胶质细胞。激活 (CD11b)、神经炎症（脑细胞因子水平）、神经变性 (FluoroJade C)、突触此外，还将研究损失（突触素）和神经发生（5-溴-2'-脱氧尿苷免疫染色）。我们还将研究脑穿透性 EPO 对血细胞比容、血清化学和组织学的影响长期给药后对主要器官的影响；目标 2：表征 cTfRMAb-EPO 融合物的影响蛋白质对 APP/PS1 小鼠认知缺陷的影响，特别是识别记忆和探索能力。使用新物体识别和旷场活动测试进行评估；目标 3：确定 cTfRMAb-EPO 融合蛋白对 APP/PS1 小鼠 Aβ 病理学的影响。将研究 EPO 融合蛋白对 Aβ 斑块负荷的影响。该提案将强调对 Aβ 斑块负荷的需求。 AD 的神经保护-神经再生方法，并提供药物递送策略，将 EPO 递送至对大脑的造血副作用可以忽略不计。