Targeting intramural von Willebrand factor (VWF) to improve vasomotor function, enhance brain parenchymal clearance, & delay development of cerebral amyloid angiopathy (CAA) in conditions of amyloid

针对壁内血管性血友病因子 (VWF) 改善血管舒缩功能，增强脑实质清除，

• 批准号: 10901009
• 负责人: Sean P Marrelli
• 金额: $ 61.93万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10901009
• 关键词: Acceleration; Age Months; Alzheimer' s Disease; Alzheimer' s disease patient; Amyloid; Amyloidosis; Arteries; Blood Platelets; Blood Vessels; Blood capillaries; Brain; Cell Culture Techniques; Cerebral Amyloid Angiopathy; Cerebrovascular Disorders; Cerebrovascular system; Cerebrum; Data; Deposition; Development; Diameter; Drainage procedure; Endothelial Cells; Endothelium; Exposure to; Fluorescence Microscopy; Functional disorder; Genetic Transcription; Hemostatic function; Histologic; Histology; Human; Hyperemia; Hypertrophy; Impaired cognition; Impairment; Incubated; Intercellular Fluid; Laser Speckle Imaging; Length; Leptomeninges; Measures; Mediating; Morphology; Mus; Muscle; Nerve; Pathologic; Pathology; Patients; Peptides; Proliferating; Protein Isoforms; Proteins; Resolution; Role; Smooth Muscle; Smooth Muscle Myocytes; Surface; Testing; Tg2576; Therapeutic; Thick; Thrombosis; Tracer; Vascular remodeling; Vasomotor; X-Ray Computed Tomography; amyloid peptide; arteriole; behavior test; brain dysfunction; brain endothelial cell; brain health; cerebral artery; cerebrovascular; extracellular; fluid flow; improved; in vivo; interstitial; knock-down; microCT; motor disorder; mouse model; multiphoton imaging; novel strategies; preservation; prevent; radiotracer; small hairpin RNA; solute; therapeutic target; transcriptome sequencing; vasomotion; von Willebrand Factor

SUMMARY. The brain vasculature contributes to the vital clearance of Ab and other solutes from brain interstitial fluid (ISF). The following proposal examines the pathological role of von Willebrand factor (VWF) within the wall of leptomeningeal arteries and arterioles, with particular focus on its potential to impair cerebrovascular function and vascular-mediated clearance in conditions of amyloidosis. VWF is expressed exclusively in endothelial cells and platelets and is widely recognized for its role in hemostasis and thrombosis. The presence of extracellular VWF in blood vessels is normally confined to the luminal endothelial surface and immediate sub-endothelial matrix. However, in certain pathological states, VWF can be found within the vascular wall (i.e. intramural VWF) where it promotes smooth muscle proliferation and wall thickening. Importantly, we and others have shown that VWF deletion profoundly prevents this vascular remodeling. Our preliminary data now expand the role of intramural VWF to conditions of AD and CAA. We show the presence of VWF within the smooth muscle region of cerebral arteries and arterioles from human AD and CAA patients and in leptomeningeal arteries (LMA) from mouse models of amyloidosis. We further show that Ab peptides promote increased VWF expression in human brain endothelial cells, supporting a contributory role for amyloidosis in VWF accumulation in the vessel wall. Based on these findings, we propose the overall hypothesis that Ab contributes to increased basolateral endothelial release of VWF, where it triggers smooth muscle proliferation and subsequent wall thickening with narrowing of leptomeningeal arteries and cortical arterioles. We further propose that the consequences of intramural VWF include cerebrovascular dysfunction, impaired brain clearance of Ab peptides, and the acceleration of cerebral artery angiopathy (CAA) and related functional consequences. We propose studies to a) define the mechanisms by which intramural VWF leads to pathological LMA remodeling and impaired Ab clearance and b) evaluate the novel strategy of reducing intramural VWF to mitigate vascular-related pathology in amyloidosis. In Aim 1, we will test the hypothesis that Ab contributes to increased basolateral endothelial release of VWF, where it triggers smooth muscle proliferation and subsequent wall thickening with narrowing of LMAs and cortical arterioles. We will use a combination of human cell culture studies and in vivo studies in TgAPP mouse models following selective VWF knockdown in brain endothelium. In Aim 2, we will test the hypothesis that reduction of brain endothelial VWF will result in increased Ab clearance by improving transendothelial and paravascular clearance mechanisms. We will evaluate selective knockdown of VWF in brain endothelium on different components of ISF flow and Ab clearance in TgAPP mice. In Aim 3, we will test the hypothesis that knockdown of brain endothelial VWF will prevent or stabilize vasomotor dysfunction and reduce vascular-driven pathology in conditions of amyloidosis. For this proof-of-concept study, we will employ behavior testing, in vivo measures of cerebrovascular function, and brain histological analyses to determine the beneficial effect of VWF knockdown in TgAPP mouse models. If successful, these studies will establish the pathological role of VWF located within the vascular wall and thus provide the initial experimental support for therapeutically targeting intramural VWF in conditions of amyloidosis.

摘要 脑血管系统有助于从脑间质液（ISF）中清除Ab和其他溶质。的 以下建议检查了软脑膜壁内血管性血友病因子（VWF）的病理作用 动脉和小动脉，特别关注其损害脑血管功能和血管介导的 清除淀粉样变性的条件。VWF仅在内皮细胞和血小板中表达，并广泛表达于血管内皮细胞。 公认其在止血和血栓形成中的作用。血管中细胞外VWF的存在通常是 局限于管腔内皮表面和直接的内皮下基质。然而，在某些病理状态下， VWF可以在血管壁内发现（即壁内VWF），在那里它促进平滑肌增殖和血管壁增生。 增厚重要的是，我们和其他人已经表明，VWF删除深刻地防止这种血管重塑。我们 初步数据现在将壁内VWF的作用扩展到AD和CAA的情况。我们发现VWF的存在 在来自人类AD和CAA患者的脑动脉和小动脉的平滑肌区域内以及在来自人类AD和CAA患者的脑动脉和小动脉的平滑肌区域内， 来自淀粉样变性小鼠模型的软脑膜动脉（LMA）。我们进一步表明，Ab肽促进增加 人脑内皮细胞中VWF的表达，支持淀粉样变性在VWF蓄积中的作用， 血管壁。基于这些发现，我们提出了总体假设，即Ab有助于增加基底外侧 VWF的内皮释放，引发平滑肌增殖和随后的管壁增厚并变窄 软脑膜动脉和皮质小动脉的分支我们进一步提出，壁内VWF的后果包括 脑血管功能障碍、Ab肽的脑清除受损和脑动脉血管病的加速 (CAA)以及相关的功能性后果。我们建议研究a）定义壁内VWF 导致病理性LMA重塑和受损的Ab清除，和B）评估减少壁内 VWF减轻淀粉样变性中血管相关病理。在目标1中，我们将检验Ab有助于 VWF的基底外侧内皮释放增加，在那里它触发平滑肌增殖和随后的壁 LMA和皮质小动脉增厚伴狭窄。我们将结合人类细胞培养研究， 脑内皮中选择性VWF敲低后TgAPP小鼠模型中的体内研究。在目标2中，我们将测试 假设脑内皮VWF降低将通过改善跨内皮细胞的Ab清除而导致Ab清除增加， 和血管旁清除机制。我们将在不同的条件下评估脑内皮中VWF的选择性敲低， TgAPP小鼠中ISF流动和Ab清除的组分。在目标3中，我们将测试大脑的敲除 内皮VWF将预防或稳定血管舒缩功能障碍，并减少血管驱动的病理学 淀粉样变性对于这项概念验证研究，我们将采用行为测试，脑血管的体内测量， 功能和脑组织学分析以确定VWF敲低在TgAPP小鼠模型中的有益作用。如果 如果成功，这些研究将确立VWF在血管壁内的病理作用，从而为研究VWF的作用机制提供理论依据。 在淀粉样变性条件下治疗靶向壁内VWF的初步实验支持。