Role of heparan sulfate in neural cell vulnerability to prions

硫酸乙酰肝素在神经细胞对朊病毒的脆弱性中的作用

• 批准号: 10684301
• 负责人: Patricia Aguilar Calvo
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684301
• 关键词: Abeta clearance; Affect; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Amyloid beta-Protein; Animal Model; Astrocytes; Binding; Brain; Brain region; Cause of Death; Cell membrane; Cell surface; Cells; Cerebellum; Chemicals; Dependence; Disease; Engineering; Environment; Extracellular Matrix; Glycocalyx; Glycoproteins; Goals; Heparan Sulfate Proteoglycan; Heparin; Heparitin Sulfate; Hippocampus; IGF Type 2 Receptor; In Vitro; Length; Lesion; Lysosomes; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Mentors; Microglia; Molecular; Molecular Conformation; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Oligodendroglia; Pathogenesis; Pathway interactions; Patients; Pattern; Persons; Physiological; Pilot Projects; Population; Post-Translational Protein Processing; PrP; Prion Diseases; Prions; Protein Isoforms; Proteins; Research; Role; Senile Plaques; Sulfate; Tail; Techniques; Testing; Training; Tropism; Variant; abeta oligomer; age related neurodegeneration; aged; cell type; design; differential expression; disease phenotype; in vivo; innovation; mannose 6 phosphate; mast cell; mimetics; misfolded protein; mouse model; neuroblastoma cell; neuroprotection; new therapeutic target; polysulfated glycosaminoglycan; prevent; protein aggregation; rational design; therapeutic target; transmission process; uptake

PROJECT SUMMARY / ABSTRACT A major unresolved question in neurodegenerative disease is the mechanisms that drive selective cell vulnerability. Heparan sulfate proteoglycans (HSPGs) are glycoproteins that promote oligomerization of amyloid- β and prions in vitro and slow the clearance of amyloid-β in the brain of an Alzheimer’s disease mouse model. HSPGs interact with misfolded proteins through their HS chains and promote their internalization in immortalized neural cells. This protein aggregate uptake is profoundly impacted by the HS length and level of sulfation which, importantly, broadly differ between cell types. We and others have used highly sulfated HS-like glycopolymers to test the crucial role of HS in the interaction and in vitro replication of prions. However, the composition of endogenous HS and their specific roles in healthy aged and disease-affected brain are unknown. I found that mice expressing shorter HS chains showed prolonged survival and profoundly altered prion plaque distribution in brain when infected with a plaque-forming prion strain, but did not show any change in the prion disease phenotype caused by aggregate-forming prions. Here I will define the HS molecules that bind to physiological and misfolded prion protein in different neuronal populations. I hypothesize that the interaction of HS with misfolded prions is a major determinant underlying the selective cell vulnerability in prion disease. In Aim 1, I will determine the role of HS sulfation in the prion replication i) in vitro, using HS isolated from distinct neuronal populations, and ii) in vivo, by mouse models deficient in HS sulfation. I will measure how the variation in the HS sulfation impacts the PrP cell tropism and lesion targets in the brain, and how age affects HS composition. I will next manipulate the HS composition in different neuronal populations i) to measure the selective cell uptake of prions strains and their degree of dependence on HS (Aim 2), and ii) to test a new strategy to block prion progression based on using HSPG mimetics as vehicles to promote prion degradation in lysosomes (Aim 3). I expect to define the molecular mechanisms underlying selective cell vulnerability in prion disease and to discover new targets for the rational design of neuroprotective therapies for patients with prion disease. Due to the many commonalities between the pathogenesis of prion disease and Alzheimer’s disease, I plan to ultimately extend my research strategy to the study of cell targeting by amyloid-β. This K99/R00 application is an ideal pathway to independence that is supported by an outstanding group of mentors and advisors, extensive training in highly innovative techniques, a world-class scientific environment, and clear departmental commitment.

项目摘要 /摘要 神经退行性疾病中的一个主要未解决的问题是驱动选择性细胞的机制 脆弱性。硫酸肝素蛋白聚糖（HSPGS）是糖蛋白，促进淀粉样蛋白的寡聚化 β和prions在体外，并减慢阿尔茨海默氏病小鼠模型大脑中淀粉样蛋白β的清除。 HSPG通过其HS链与错误折叠的蛋白质相互作用，并在永生中促进其内在化 神经细胞。这种蛋白质骨料的摄取受到HS长度和硫酸水平的深刻影响，硫酸盐的水平和水平。 重要的是，细胞类型之间的差异很大。我们和其他人使用了高度硫酸的HS样糖聚合物 测试HS在prions的相互作用和体外复制中的关键作用。但是， 内源性HS及其在健康老年和受疾病影响的大脑中的特定作用尚不清楚。我发现 表达较短的HS链的小鼠显示长期生存和深刻改变的Prion斑块分布 在大脑中感染了形成斑块的pr菌菌株时，但没有显示出病毒疾病的任何变化 由骨料形成的王室引起的表型。在这里，我将定义与生理结合的HS分子 并在不同的神经元种群中折叠率错误。我假设HS与 错误折叠的王室是prion病中选择性细胞脆弱性的主要决定因素。 AIM 1，我将使用从不同的HS中确定HS硫酸化在Prion复制中的作用 神经元种群和II）在体内，由小鼠模型缺乏HS硫酸化。我将衡量如何变化 在HS中，硫酸盐会影响大脑中的PRP细胞对流和病变靶标，以及年龄如何影响HS 作品。接下来，我将操纵不同神经元种群中的HS组成i）测量 选择性细胞吸收prions菌株及其对HS的依赖程度（AIM 2），ii）测试新策略 基于使用HSPG Mimetics作为促进prion降解的工具来阻止prion进展 溶酶体（AIM 3）。我希望定义prion中选择性细胞脆弱性的分子机制 疾病并发现针对Prime患者的神经保护疗法合理设计的新目标 疾病。由于病毒疾病的发病机理和阿尔茨海默氏病之间有许多共同点， 计划最终将我的研究策略扩展到淀粉样蛋白β对细胞靶向的研究。此K99/R00 应用是一个理想的独立途径，得到了一群杰出的导师和 顾问，高度创新技术，世界一流的科学环境的广泛培训，清晰 部门承诺。