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Virus nanoparticles as autophagy activators

作为自噬激活剂的病毒纳米粒子

基本信息

批准号：
8669826
负责人：
Laura Segatori
金额：
$ 21.75万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8669826
关键词：
Appearance Autophagocytosis Autophagosome Behavior Biogenesis Biology Blood - brain barrier anatomy Capsid Cell Nucleus Cells Clear Cell Data Defective Viruses Dependovirus Deposition Development Disease Endocytosis Engineering Fibroblasts Functional disorder Goals Human Huntington Disease In Vitro Infection Knowledge Lead Link Lipofuscin Lysosomes Mediating Molecular Nerve Degeneration Neuraxis Neurodegenerative Disorders Neuronal Ceroid-Lipofuscinosis Organelles Outcome Parkinson Disease Pathway interactions Patients Positioning Attribute Process Property Proteolipids Public Health Regulation Regulator Genes Reporting Research Research Personnel Role Serotyping Shapes System Technology Testing Therapeutic Translational Research Variant Viral Genome Virus Work activating transcription factor base design effective therapy gene therapy in vitro Model innovation nanomaterials nanoparticle particle performance tests prevent protein aggregate protein misfolding prototype public health relevance response skills synuclein therapeutic development therapeutic target transcription factor uptake wasting

项目摘要

DESCRIPTION (provided by applicant): Autophagy mediates degradation of cytoplasmic material through the lysosomes, and inefficiencies in this vital homeostatic process result in devastating diseases. For example, neurodegenerative lysosomal storage disorders are characterized by deficiencies in lysosomal degradation and autophagy dysfunction. Inefficient autophagy also leads to accumulation of aggregated proteins and neurodegeneration in protein misfolding diseases such as Parkinson's disease. Thus, translational research strategies to activate autophagy could dramatically impact the development of therapeutics for a large range of neurodegenerative diseases. Interestingly, evidence of autophagy induction by nanomaterials has recently emerged. Unfortunately, most current synthetic nanoparticle systems are unable to cross the blood-brain barrier (BBB), precluding their use to treat the central nervous system. In addition, most nanomaterials disrupt lysosomal function, ultimately leading to block of autophagy flux and impaired clearance. To overcome these limitations, we propose to develop a platform of virus nanoparticles (VNP) that can induce the coordinated activation of autophagy and lysosomal biogenesis. Our VNP technology is based on the adeno-associated virus (AAV), and will particularly focus on AAV serotype 9 that was shown to cross the BBB. Evidence of the integrated and co-regulated roles of lysosomes and autophagosomes emerged from the recent discovery of a master regulator of autophagy and lysosome biogenesis, the transcription factor EB (TFEB). The long-term goal of this work is the development of a VNP-based platform technology to activate TFEB and enhance clearance of autophagic cargo by lysosomes. The objective of this proposal is to identify the design rules for generating VNPs that can activate TFEB and enhance clearance in vitro. The central hypothesis of this study, based on our pilot data, is that the TFEB-activating properties of VNPs depend on cellular uptake of the particle and not on the infectivity of the capsid since a defective VNP unable to productively infect cells can still activate TFEB. If successful, the proposed research will produce an enabling therapeutic platform for the treatment of neurodegenerative diseases characterized by accumulation of autophagic substrates. Specifically, we propose to define the design rules for building VNP-based activators of TFEB (aim 1), and we will test the performance of VNPs in vitro by evaluating clearance of i) lipofuscin in fibroblasts derived from patients with Neuronal Ceroid Lipofuscinosis and ii) aggregated ¿-synuclein in neuroglioma cells (aim 2). The proposed research is significant because it will generate a therapeutic platform able to promote co-regulated activation of the lysosome-autophagy system for effective treatment of neurodegenerative diseases. This approach is innovative because it promises to overcome limitations of currently available synthetic nanomaterials that cannot cross the BBB. Results from this study will also significantly advance our knowledge of AAV biology, as no scientific information is currently available concerning the impact of AAV on the autophagy pathway.

描述(申请人提供)：自噬通过溶酶体介导胞浆物质的降解，而在这一重要的体内平衡过程中效率低下会导致毁灭性的疾病。例如，神经退行性溶酶体储存障碍的特征是溶酶体降解不足和自噬功能障碍。低效的自噬还会导致聚集蛋白的积累和蛋白质错误折叠疾病(如帕金森氏病)的神经退化。因此，激活自噬的翻译研究策略可能会极大地影响大范围神经退行性疾病的治疗方法的发展。有趣的是，最近出现了纳米材料诱导自噬的证据。不幸的是，目前的大多数合成纳米粒子系统无法通过血脑屏障(BBB)，因此无法用于治疗中枢神经系统。此外，大多数纳米材料扰乱了溶酶体的功能，最终导致自噬通量的阻断和清除的损害。为了克服这些限制，我们建议开发一个病毒纳米颗粒(VNP)平台，它可以诱导自噬和溶酶体生物发生的协调激活。我们的VNP技术是以腺相关病毒(AAV)为基础的，将特别专注于被证明跨越血脑屏障的AAV 9型。最近发现了自噬和溶酶体生物发生的主要调节因子--转录因子EB(TFEB)，证明了溶酶体和自噬的整合和共同调节作用。这项工作的长期目标是开发一种基于VNP的平台技术，以激活TFEB并加强溶酶体对自噬货物的清除。这项建议的目的是确定产生VNPs的设计规则，以激活TFEB并提高体外清除率。这项研究的中心假设是，基于我们的试点数据，VNPs的TFEB激活特性取决于细胞对颗粒的摄取，而不是取决于衣壳的感染性，因为无法高效感染细胞的有缺陷的VNP仍然可以激活TFEB。如果成功，这项拟议的研究将为治疗以自噬底物积累为特征的神经退行性疾病提供一个有效的治疗平台。具体地说，我们建议定义构建基于VNP的TFEB激活剂的设计规则(Aim 1)，并将通过评估i)神经样脂褐质沉积症患者来源的成纤维细胞中的脂褐素和ii)神经胶质瘤细胞中聚集的突触核蛋白(Aim 2)的清除来测试VNPs的体外性能。这项拟议的研究具有重要意义，因为它将产生一个治疗平台，能够促进溶酶体-自噬系统的共同调节激活，以有效治疗神经退行性疾病。这种方法是创新的，因为它承诺克服目前可用的合成纳米材料的限制，这些材料不能越过BBB。这项研究的结果也将极大地促进我们对AAV生物学的了解，因为目前还没有关于AAV对自噬途径的影响的科学信息。