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.

项目摘要/摘要 对认知障碍和痴呆（VCID）的血管贡献包括脑血管谱 变化参见大脑小血管疾病，包括脑淀粉样血管病（CAA），其中 病理淀粉样β（Aβ）蛋白在脑毛细血管和动脉的壁上积聚。随着增加 年龄，并且患有早期寿命的高脂血症，沿着大脑壁的Aβ失败 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 沿毛细血管和动脉的细胞外基质（ECM）组合的基底膜 周围排水（iPad）。衰老，除其他因素外，还会改变ECM的结构，从而导致 AβiPad的失败。此外，脑血管重塑和血管生成可能早期代表 VCID中血流量减少的补偿性变化，并部分通过增加蛋白水解发生 周围ECM的营业额，例如Perlecan。我们已经证明了perlecan域V（DV）蛋白 大大降低了淀粉样蛋白的毒性，同时增强了血管生成。人CAA患者缺乏perlecan 仅在淀粉样蛋白阳性vissel上染色。这些研究共同提供了打击证据来提出关键 perlecan在脑血管Aβ沉积及其相关的有害作用中的作用。最后，蛋白聚糖 可以对内皮转运蛋白机制（例如P-糖蛋白（P-gp））产生影响，这些机制对Aβ至关重要 从大脑中消除，这也随着年龄的增长而失败。 iPad上的燃烧增加与血脑有关 障碍（BBB）破坏，最终使大脑中的Aβ沉积担心，这导致了VCID。作为 脑血管基底机制随着年龄的增长而变化，对ECM的干预措施改善了iPad 可以代表一种早期，批判性和治疗性相关的方法，用于预防CAA和 VCID。在这里，我们假设DV在Aβ的血管清除中起重要的内源作用 历史上可以在VCID中与iPad相关的变化中进行利用。为了支持这一假设， 初步数据表明：1。DV通过增加P-gp的表达和活性来增加HAβ的清除率 体外脑微血管，2。DV在体外跨WT小鼠衍生的BEC中增加了HAβ的转运，3。小鼠 明示正常perlecan和DV水平的10％（称为PLN - / - ）已确定的iPad受损 在其壁上有Aβ的毛细血管和动脉数量较高（与注射Aβ的WT对照相比） 立体定向注射Aβ和4。PLN - / - 衍生的BEC具有不足的Aβ传输，可以通过DV挽救 体外治疗。通过这些结果，我们建议分析DV介导的Aβ清除率的增加 在机械细节中跨越脑内皮细胞，并确定DV在 Aβ和实验性CAA的iPad。