Inhibition of Amyloid Formation by Heterogeneous Nanoparticles with Chaperone-like Activity

具有伴侣样活性的异质纳米颗粒抑制淀粉样蛋白形成

• 批准号: 9901460
• 负责人: Joel Kaar
• 金额: $ 17.9万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9901460
• 关键词: Address; Aging; Alzheimer' s Disease; Alzheimer' s disease patient; Amyloid; Amyloid beta-Protein; Amyloid fibers; Binding; Biological Assay; Chemicals; Circular Dichroism; Creutzfeldt-Jakob Syndrome; Disease; Fiber; Fibronectins; Fluorescence; Fluorescence Resonance Energy Transfer; Future; Glycerol; Kinetics; Lead; Link; Lipid Bilayers; Literature; Malignant Neoplasms; Measurement; Measures; Mediating; Methods; Modeling; Molecular; Molecular Chaperones; Monitor; Mutation; Nitroreductases; Parkinson Disease; Pathogenesis; Peptides; Phosphorylcholine; Polymers; Presenile Alzheimer Dementia; Protein Dynamics; Proteins; Silicon Dioxide; Structure; Surface; Testing; Therapeutic; Work; age related; amyloid fibril formation; amyloid formation; amyloid structure; antibody engineering; beta pleated sheet; biophysical techniques; ethylene glycol; in vivo; innovation; insight; interest; misfolded protein; mutant; nanomaterials; nanoparticle; novel; novel therapeutic intervention; preclinical study; prevent; protein aggregation; protein folding; protein misfolding; protein structure; single molecule; small molecule; success

SUMMARY The formation of amyloid fibrils due to protein aggregation represents a central step in the pathogenesis of many age-related diseases for which there are no current cures. At the core of this step is the misfolding of one or more key proteins, which is converted from a natively folded or unfolded state to an aggregation-prone state that is rich in b-sheet structure. While there have been intensive efforts to develop therapeutic approaches to inhibit amyloid fiber formation during its initial stages, such strategies have met with only limited success. As such, there remains a need for alternative strategies to mediate even earlier states of amyloid oligomerization by re- folding and thus rescuing misfolded proteins prior to aggregating. The overall aim of this proposal is to develop nanomaterials with chaperone-like activity to promote re-folding of misfolded proteins and thus have therapeutic potential to inhibit amyloid formation. Specifically, in this approach, we propose to rationally modify nanoparticles with novel coatings that stabilize the native structure of amyloid-associated proteins and can be delivered in vivo. Of particular interest will be investigating the extent to which chemically heterogeneous polymer brushes and mixed lipid bilayers promote re-folding of the protein amyloid-b (Ab42), which is a precursor to amyloid fibers that are formed in Alzheimer’s disease. In support of this approach, we have previously shown that random co- polymer brushes composed of poly(ethylene glycol) and poly(sulfobetaine) as well as mixed supported lipid bilayers compared of varying ratios of 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3- phospho-(1'-rac-glycerol) both stabilized and promoted the re-folding of model proteins, including fibronectin and nitroreductase. Using coatings made of such materials, we will test the central hypothesis that nanoparticles coated with dynamic and heterogeneous layers can reduce the conversion of Ab42 to its aggregation-prone state and prevent amyloid formation via a chaperone-like mechanism. In line with this hypothesis, the specific aims of this proposal are to: (1) elucidate the mechanism and optimize the composition of heterogeneous coatings for Ab42 stabilization by nanoparticles via a chaperone-like mechanism (Aim 1) and (2) correlate Ab42 stabilization by chemically heterogeneous nanoparticles with the inhibition amyloid formation (Aim 2). The chaperone-like activity of the nanoparticle coatings will be characterized quantitatively using single-molecule biophysical methods that are uniquely sensitive to monitoring protein structure and dynamics (e.g., re-folding) on surfaces. To correlate the stabilization of Ab42 with the inhibition of amyloid formation, the impact of the coatings on the formation of amyloid fibrils will be measured in the presence of coated and uncoated nanoparticles while varying coating composition. Additionally, the utility of the coated nanoparticles in inhibiting the aggregation of Ab42 and Ab40 as well as Ab42 with mutations associated with Alzheimer’s disease (i.e., E22G) will also be measured. These studies will provide a complete molecular picture of this approach, as well as a general understanding that may be applied to the rescue of proteins implicated in other protein misfolding diseases, including cancer.

总结 由于蛋白质聚集而形成淀粉样纤维代表了许多疾病发病机制的中心步骤， 与年龄有关的疾病，目前还没有治愈方法。这一步的核心是一个或多个基因的错误折叠， 更多的关键蛋白质，其从天然折叠或未折叠状态转化为易于聚集的状态， 富含B片层结构。虽然已经进行了大量的努力来开发治疗方法以抑制 在淀粉样蛋白纤维形成的初始阶段，这种策略只取得了有限的成功。因此，在本发明的一个方面， 仍然需要替代的策略来通过再聚合来介导淀粉样蛋白寡聚化的甚至更早的状态， 折叠并因此在聚集之前拯救错误折叠的蛋白质。本提案的总体目标是发展 具有分子伴侣样活性的纳米材料，以促进错误折叠的蛋白质的再折叠， 抑制淀粉样蛋白形成的潜力。具体来说，在这种方法中，我们建议合理地修饰纳米颗粒， 其具有稳定淀粉样蛋白相关蛋白的天然结构的新型涂层，并且可以在体内递送。 特别感兴趣的将是调查化学非均相聚合物刷的程度， 混合脂质双层促进淀粉样蛋白-b（Ab 42）的再折叠，其是淀粉样纤维的前体 在阿尔茨海默氏症中形成。为了支持这种方法，我们以前已经表明，随机共- 由聚乙二醇和聚磺基甜菜碱组成的聚合物刷以及混合支持的脂质 不同比例的1，2-二油酰基-sn-甘油基-3-磷酸胆碱和1，2-二油酰基-sn-甘油基-3-磷酸胆碱的双层比较 磷酸-（1 '-外消旋-甘油）既稳定又促进模型蛋白的重折叠，包括纤连蛋白和 硝基还原酶使用这种材料制成的涂层，我们将测试中心假设， 用动态和非均匀层包覆可以减少Ab 42向其聚集倾向状态的转化 并通过类似伴侣的机制防止淀粉样蛋白的形成。根据这一假设， 本文的主要目的是：（1）阐明非均相涂层的形成机理，优化非均相涂层的组成， 纳米颗粒通过分子伴侣样机制稳定Ab 42（目的1）和（2）关联Ab 42稳定 通过化学异质纳米颗粒与抑制淀粉样蛋白的形成（目的2）。监护人般的 纳米颗粒涂层的活性将使用单分子生物物理方法定量表征。 对监测蛋白质结构和动力学独特敏感的方法（例如，再折叠）。 为了将Ab 42的稳定化与淀粉样蛋白形成的抑制相关联，研究了包衣对Ab 42的稳定化的影响。 淀粉样蛋白原纤维的形成将在包衣和未包衣的纳米颗粒存在下测量 涂料组合物。此外，包被的纳米颗粒在抑制Ab 42和Ab 43聚集中的效用也被证实。 Ab 40以及具有与阿尔茨海默病相关的突变的Ab 42（即，E22 G）也将被测量。 这些研究将提供一个完整的分子图片这种方法，以及一般的理解 其可应用于拯救与其它蛋白质错误折叠疾病（包括癌症）有关的蛋白质。

项目成果

Mixed Phospholipid Vesicles Catalytically Inhibit and Reverse Amyloid Fibril Formation.

• DOI: 10.1021/acs.jpclett.0c02074
• 发表时间: 2020-09-03
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY LETTERS
• 影响因子: 5.7
• 作者: Chaparro Sosa, Andres F.;da Silva, Sabrina Matos de Oliveira;Morgan, Garry P.;Schwartz, Daniel K.;Kaar, Joel L.
• 通讯作者: Kaar, Joel L.

Tuning the surface charge of phospholipid bilayers inhibits insulin fibrilization.

• DOI: 10.1016/j.colsurfb.2022.112904
• 发表时间: 2022-12
• 期刊: Colloids and surfaces. B, Biointerfaces