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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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.

项目概要/摘要神经退行性疾病中一个尚未解决的主要问题是驱动选择性细胞的机制硫酸乙酰肝素蛋白聚糖（HSPG）是促进淀粉样蛋白寡聚化的糖蛋白。 β 和朊病毒在体外并减缓阿尔茨海默病小鼠模型大脑中β淀粉样蛋白的清除。 HSPG 通过其 HS 链与错误折叠的蛋白质相互作用，并促进其在永生化中的内化这种蛋白质聚集体的摄取受到 HS 长度和硫酸化水平的深刻影响，我们和其他人使用了高度硫酸化的类 HS 糖聚合物。测试 HS 在朊病毒相互作用和体外复制中的关键作用。我发现内源性 HS 及其在健康老年人和受疾病影响的大脑中的具体作用尚不清楚。表达较短 HS 链的小鼠表现出更长的生存期和显着的朊病毒斑块分布当感染形成斑块的朊病毒株时，大脑中出现了这种变化，但朊病毒病没有表现出任何变化由形成聚集体的朊病毒引起的表型在这里我将定义与生理学结合的 HS 分子。以及不同神经元群体中错误折叠的朊病毒蛋白，我利用了 HS 的相互作用。错误折叠的朊病毒是朊病毒疾病中选择性细胞脆弱性的主要决定因素。目标 1，我将确定 HS 硫酸化在朊病毒复制中的作用 i) 在体外，使用从不同的神经元群体，以及 ii) 体内，通过缺乏 HS 硫酸化的小鼠模型，我将测量变化如何。 HS 硫酸化影响大脑中 PrP 细胞的趋向性和病变目标，以及年龄如何影响 HS 接下来我将操纵不同神经群体中的 HS 组成 i) 来测量细胞对朊病毒株的选择性摄取及其对 HS 的依赖程度（目标 2），以及 ii) 测试新策略基于使用 HSPG 模拟物作为载体来阻止朊病毒进展，以促进朊病毒降解我希望定义朊病毒选择性细胞脆弱性的分子机制。疾病并发现合理设计朊病毒患者神经保护疗法的新靶点由于朊病毒病和阿尔茨海默病的发病机制有许多共同点，我认为计划最终将我的研究策略扩展到淀粉样蛋白-β 的细胞靶向研究。申请是获得独立的理想途径，并得到优秀导师和团队的支持顾问、高度创新技术的广泛培训、世界一流的科学环境以及清晰的部门承诺。