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

• 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
• 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' s Disease; Alzheimer' 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

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.

项目总结/摘要 目前建议的目标是开发一种脑穿透性促红细胞生成素（EPO）类似物， 造血系统副作用EPO是一种独特的治疗AD的候选药物，因为它靶向一系列的 参与AD的病理生理学和疾病进展的过程，重要的是， 神经发生和改善认知。这明显不同于许多抗A β药物， 目前正在开发用于AD，因为抗A β药物不能逆转现有的神经元损伤， 认知能力下降另一方面，EPO具有神经保护和神经再生特性。这 为开发EPO治疗AD提供了科学依据。EPO发展的两大障碍 AD是：a）有限的血脑屏障（BBB）穿透，需要经颅施用或使用高浓度的 剂量，和B）高剂量的EPO导致不希望的造血作用。为了实现我们的目标， EPO蛋白与抗小鼠转铁蛋白受体（TfR）的单克隆抗体（MAb）的结合， 工程化并命名为cTfRMAb-EPO。cTfRMAb-EPO融合蛋白提供了双重优点： 它通过BB B TfR迅速进入脑，和B）它通过 外周TfR，导致可忽略的造血作用。我们的中心假设是， 施用cTfRMAb-EPO融合蛋白改变AD病理生理学的多个靶点， 疾病进展（Aβ-病理学（↓）、神经炎症（↓）、突触丢失（↓）、神经发生（↑）、认知 功能（↑），因此在AD中具有治疗性，造血作用可忽略不计。在拟议的研究中， APPswe，PSEN 1dE 9（APP/PS1）小鼠将用cTfRMAb-EPO融合蛋白、重组人红细胞生成素（EPO）或重组人红细胞生成素（EPO）处理。 人EPO（rhuEPO）或载体，并且将解决以下具体目的：目的1：确定 cTfRMAb-EPO融合蛋白在APP/PS1小鼠中的细胞保护作用。特别是小胶质细胞 活化（CD 11b）、神经炎症（脑细胞因子水平）、神经变性（FluoroJade C）、突触 将研究突触素的丢失和神经发生（5-溴-2 ′-脱氧尿苷免疫染色）。此外，本发明还 我们还将研究穿透脑的EPO对红细胞压积、血清化学和组织学的影响。 慢性给药后对主要器官的作用;目的2：表征cTfRMAb-EPO融合物的作用 蛋白质对APP/PS1小鼠认知缺陷的影响。特别是，识别记忆和探索将 使用新物体识别和旷场活动测试进行评估;目的3：确定 cTfRMAb-EPO融合蛋白对APP/PS1小鼠Aβ病理学的影响。具体而言，cTfRMAb- 将研究EPO融合蛋白对Aβ斑块负荷的影响。该提案将强调需要 神经保护-神经再生方法，并提供一种药物递送策略，将EPO递送至 对大脑的造血副作用可以忽略不计。