Perlecan Domain V as a therapeutic VCID strategy for the clearance of amyloid beta from the brain in cerebral amyloid angiopathy

Perlecan 结构域 V 作为治疗性 VCID 策略，用于清除大脑淀粉样血管病中的β淀粉样蛋白

• 批准号: 10372826
• 负责人: Gregory Jaye Bix
• 金额: $ 38.26万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372826
• 关键词: ABCB1 gene; ATP phosphohydrolase; Abeta clearance; Age; Age of Onset; Age-Months; Aging; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Arteries; Basement membrane; Binding; Biological Assay; Blood Vessels; Blood capillaries; Blood flow; Brain; Brain Diseases; C-terminal; Cerebral Amyloid Angiopathy; Cerebrovascular system; Cerebrum; Clinical; Cognition; Cognitive; Collagen Type IV; Data; Dementia; Disease; Drainage procedure; Electron Microscopy; Endothelium; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Extracellular Structure; Failure; Female; Fibronectins; Fluorometry; Genotype; Health; Heparan Sulfate Proteoglycan; Histologic; Human; Hyperlipidemia; Hypertension; Impaired cognition; Impairment; In Vitro; Injections; Integrin alpha5beta1; Intercellular Fluid; Intervention; Laminin; Lead; Mediating; Memory; Microvascular Dysfunction; Morphology; Mus; Pathogenesis; Pathologic; Patients; Pattern; Play; Prevention; Prevention approach; Process; Proteins; Proteoglycan; Role; Small Interfering RNA; Smooth Muscle Myocytes; Stains; Study models; System; Therapeutic; Time; Toxic effect; Vascular Dementia; Vascular blood supply; Western Blotting; abeta accumulation; abeta deposition; aged; angiogenesis; apolipoprotein E-4; arteriole; blood-brain barrier disruption; brain endothelial cell; cerebral artery; cerebral capillary; cerebral microvasculature; cerebrovascular; cerebrovascular amyloid; hypoperfusion; improved; in vitro Model; inhibitor; male; middle age; monolayer; mouse model; neurovascular; novel; object recognition; perlecan; proto-oncogene protein pim-1; vascular cognitive impairment and dementia

PROJECT SUMMARY/ABSTRACT Vascular contributions to cognitive impairment and dementia (VCID) encompass a spectrum of cerebrovascular changes seen in small vessel diseases of the brain including cerebral amyloid angiopathy (CAA) where pathologic amyloid beta (Aβ) protein accumulates in the walls of cerebral capillaries and arteries. With increasing age, and with hyperlipidemia in early to mid-life, there is a failure of clearance of Aβ along the walls of cerebral capillaries and arteries resulting in CAA. One of the key mechanisms for the elimination of Aβ from the brain is along the extracellular matrix (ECM)-composed basement membranes of capillaries and arteries, as intramural periarterial drainage (IPAD). Aging, among other factors, changes the structure of the ECM, resulting in the failure of IPAD of Aβ. Furthermore, cerebrovascular remodeling and angiogenesis may represent early compensatory changes to the reduced blood flow seen in VCID and occur in part by increasing the proteolytic turnover of the surrounding ECM such as perlecan. We have demonstrated that perlecan domain V (DV) protein greatly reduces amyloid toxicity while enhancing angiogenesis. Human CAA patients showed a lack of perlecan staining only on amyloid positive vessels. Collectively, these studies offer striking evidence to suggest a critical role of perlecan in cerebrovascular Aβ deposition and its associated deleterious effects. Lastly, proteoglycans can have effects on endothelium transporter mechanisms such as P- glycoprotein (P-gp) which are vital to Aβ elimination from the brain, which also fail with age. This increased burden on IPAD correlates to blood-brain barrier (BBB) disruption, ultimately worsening Aβ deposition in the brain, which contributes to VCID. As cerebrovascular basement membranes change with age, interventions upon the ECM that improve IPAD may represent an early, critical, and therapeutically relevant approach for the prevention of CAA and VCID. Here we hypothesize that DV plays an important endogenous role in vascular clearance of Aβ which can be therapeutically exploited in IPAD-related changes in VCID. In support of this hypothesis, preliminary data suggest that: 1. DV increases hAβ clearance by increasing P-gp expression and activity in cerebral microvessels in vitro, 2. DV increases hAβ transit across WT mice-derived BECs in vitro, 3. mice that express 10% of total normal perlecan and DV levels (referred to as Pln-/-) have impaired IPAD as determined by a higher number of capillaries and arterioles with Aβ in their walls (compared to Aβ-injected WT controls) after stereotactic injection of Aβ, and 4. Pln-/- - derived BEC’s have deficient Aβ transit that can be rescued by DV treatment in vitro. Armed with these results, we propose to analyze DV-mediated increased clearance of Aβ across brain endothelial cells in mechanistic detail, and to determine the role and therapeutic potential of DV on IPAD of Aβ and experimental CAA.

项目总结/摘要 血管对认知障碍和痴呆（VCID）的贡献包括一系列脑血管疾病， 在脑小血管疾病中观察到的变化，包括脑淀粉样血管病（CAA）， 病理性淀粉样β（Aβ）蛋白积聚在脑毛细血管和动脉壁中。随着增加 年龄越大，高脂血症越早，Aβ沿着脑血管壁的清除率越低， 毛细血管和动脉导致CAA。从大脑中消除Aβ的关键机制之一是 沿着细胞外基质（ECM）组成的毛细血管和动脉的基底膜，如壁内 动脉周围引流（iPad）。除其他因素外，老化会改变ECM的结构，导致 Aβ的iPad失效。此外，脑血管重塑和血管生成可能代表早期 在VCID中观察到的对减少的血流量的代偿性变化，部分是通过增加蛋白水解而发生的。 周围ECM如串珠素的周转。我们已经证明，串珠蛋白结构域V（DV）蛋白 大大降低淀粉样蛋白毒性，同时增强血管生成。人类CAA患者显示缺乏串珠素 仅在淀粉样蛋白阳性血管上染色。总的来说，这些研究提供了惊人的证据，表明一个关键的 串珠素在脑血管Aβ沉积及其相关有害作用中的作用最后，蛋白聚糖 可影响内皮转运机制，如对Aβ至关重要的P-糖蛋白（P-gp） 从大脑中清除，这也随着年龄的增长而失效。iPad上的这种增加的负担与血脑 血脑屏障（BBB）破坏，最终恶化脑中的Aβ沉积，导致VCID。作为 脑血管基底膜随年龄变化，对ECM进行干预可改善iPad 可能代表了预防CAA的早期、关键和治疗相关的方法， VCID。在此我们假设DV在Aβ的血管清除中起重要的内源性作用 其可以在iPad相关的VCID变化中被治疗性地利用。为了支持这一假设， 初步数据表明：1. DV通过增加P-gp表达和活性增加hAβ清除率， 体外脑微血管; 2. DV在体外增加hAβ穿过WT小鼠来源的BEC的转运，3.的小鼠 表达10%的总正常串珠素和DV水平（称为Pln-/-）具有受损的iPad， 注射A β后，血管壁中含有Aβ的毛细血管和小动脉数量增加（与注射Aβ的WT对照组相比）， 立体定向注射Aβ; Pln-/-衍生的BEC具有可被DV拯救的缺陷Aβ转运 体外治疗根据这些结果，我们建议分析DV介导的Aβ清除率增加 通过脑内皮细胞的机制细节，并确定DV的作用和治疗潜力， Aβ的iPad和实验性CAA。