Development of Small Heat Shock Proteins as Therapeutic Agents in the Eye

开发小热激蛋白作为眼部治疗剂

• 批准号: 7833268
• 负责人: Jonathan Mark Petrash
• 金额: $ 49.92万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7833268
• 关键词: Accounting; Address; Affect; Age related macular degeneration; Animal Model; Area; Autoimmune Diseases; Blindness; Cataract; Cells; Clinic; Complex; Crystallins; Degenerative Disorder; Development; Diagnosis; Disease; Disease model; Drug Formulations; Effectiveness; Encapsulated; Etiology; Eye; Eye diseases; Genes; Glaucoma; Goals; Half-Life; Heat shock proteins; Hereditary Disease; Human; Injection of therapeutic agent; Knockout Mice; Medical; Metabolic stress; Methods; Modeling; Molecular Chaperones; Molecular Weight; Nerve Degeneration; Pathogenesis; Pathology; Pathway interactions; Patients; Penetration; Peptide Hydrolases; Pharmaceutical Preparations; Plasmids; Productivity; Protein Family; Protein Subunits; Proteins; Quality of life; Rattus; Retinitis Pigmentosa; Rhodopsin; Risk; Series; Stress; System; Technology; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Transgenic Organisms; Translational Research; Viral; abstracting; base; cost; expression vector; improved; lens; macromolecule; member; molecular size; mouse model; mutant; nanomedicine; nanoparticle; novel; prevent; protein aggregate; protein aggregation; protein complex; protein misfolding; public health relevance; targeted delivery; therapeutic protein; tissue culture; tool; treatment strategy

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (15) Translational Science and specific Challenge Topic, 15-EY-101: Protein misfolding in degenerative diseases of the eye. Identifying therapeutic pharmacological agents/drugs that prevent the misfolding/aggregation of proteins could provide new tools for treating these diseases. Although degeneration of the cells in the eye underlies various pathologies including cataracts, age related macular degeneration and glaucoma, currently there are no approved therapeutic agents for treating cellular degeneration associated with these disorders. One difficulty underlying the development of such therapeutic agents is the complex etiology of these degenerative disorders, wherein multiple cellular pathways are altered. Thus, a therapeutic candidate would ideally target multiple stress pathways underlying cellular degeneration. Our project addresses a critical dilemma for protein-based therapy in the eye: how to provide therapeutic proteins for long-term therapy while at the same time minimizing risks and costs associated with repeated intraocular injections. It is likely that drugs intended for treating degenerative disorders of the eye should be administered from the time of diagnosis onwards. Currently approved protein therapeutics administered in the eye have a relatively short half-life (<10 days), which necessitates repeated injections to the eye approximately every 6 weeks. Thus, there is a need for slow release systems for prolonged delivery of macromolecules to the eye. Technology for targeted delivery of macromolecules to degenerating cells of the eye is also an important goal. Protein drugs, besides being susceptible to proteolytic enzymes that are ubiquitous, do not enter target cells efficiently due to their large molecular size. In order to overcome this limitation, functionalized nanoparticles encapsulating the protein drugs need to be developed. The small heat shock proteins (sHSP) proteins aA-crystallin and aB-crystallin have excellent potential as therapeutic proteins against some of the most common eye diseases seen in the clinic today. Like other members of the sHSP family, ¿-crystallin oligomers display chaperone-like activity defined by their ability to prevent aggregation of other proteins. Outside the eye, ¿-crystallin has been demonstrated to co-localize with protein aggregates implicated in the pathogenesis of a variety of diseases, including fibrillary bundles associated with neurodegenerative and autoimmune diseases. Introduction of sHSPs via plasmid or viral expression vectors has been demonstrated to rescue phenotypic markers in a variety of disease models, including retinitis pigmentosa (RP) and aggregation-prone cataract mutants. Consistent with the emphasis on translational science in this challenge announcement, we propose to develop sHSP-based therapeutics to prevent and/or reverse the formation of aggregates implicated in the pathogenesis of cataract and RP. Our overarching hypothesis is that developing an efficient method for delivery of sHSPs to the eye will provide a novel new strategy for treatment of a broad range of eye conditions caused by genetic disorders and metabolic stress. Our proposal is organized around two major goals. In the first aim, we will develop and optimize various technologies for preparing nanoparticles loaded with human sHSP. In the second aim, we will test the effectiveness of sHSP delivered via nanoparticles at suppressing and/or reversing high molecular weight aggregates formed from mutant proteins associated with RP and cataracts. Specific Aim 1: Develop a series of sHSP nanomedicine formulations and optimize for sustained slow release into the eye. We will pursue four different technologies for generating slow release of sHSP complexes into the eye. Combinations of wild type and "activated" sHSP subunits derived from human sHSP genes will be incorporated into nanoparticles. Studies will be carried out to optimize protein release and tissue penetration from different nanoparticle formulations. Specific Aim 2: Test the hypothesis that sHSP can suppress formation of insoluble high molecular weight oligomeric complexes initiated by aggregation-prone rhodopsin and lens crystallin mutants. Nanoparticle formulations that demonstrate efficacy in tissue culture models of RP and cataract will be tested in animal models of protein aggregation disease, including the P23H transgenic rat model of RP and the aA-crystallin knock out mouse model of cataract. PUBLIC HEALTH RELEVANCE: This project seeks to develop new drugs to prevent or reverse blinding diseases such as cataract and retinitis pigmentosa, which are associated with aggregation of proteins. Protein aggregation diseases account for some of the most common degenerative diseases of the eye. Our proposed studies could substantially improve the quality of life of affected patients by preventing vision loss and associated financial burdens due to lost productivity and increased medical expenses.

描述（由应用程序提供）：此应用程序解决了广泛的挑战领域（15）转化科学和特定挑战主题，15-ey-101：眼睛退行性疾病中的蛋白质不折叠。识别防止蛋白质错误折叠/聚集的治疗药物/药物可以提供治疗这些疾病的新工具。 尽管眼睛中细胞的变性是各种病理的基础，包括白内障，与年龄相关的黄斑变性和青光眼，目前尚无批准的治疗剂来治疗与这些疾病相关的细胞变性。这种治疗剂的发展的一个困难是这些退化性疾病的复杂病因，其中多个细胞途径发生了改变。这是理想情况下，治疗候选者将靶向细胞变性的多个应力途径。 我们的项目解决了眼睛中基于蛋白质治疗的严重困境：如何为长期治疗提供治疗蛋白，同时最大程度地减少与重复的眼内注射相关的风险和成本。从诊断开始时，应服用用于治疗眼睛退行性疾病的药物。目前在眼中服用的蛋白质疗法的半衰期相对较短（<10天），大约每6周大约每6周重复注射。这是需要缓慢释放的系统来长时间递送大分子的眼睛。将大分子靶向分配到眼睛退化细胞的技术也是一个重要目标。蛋白质药物除了容易受到无处不在的蛋白水解酶的影响外，由于它们的大分子大小，不能有效地进入靶细胞。为了克服这一局限性，需要开发包裹蛋白质药物的功能化纳米颗粒。 小型热休克蛋白（SHSP）蛋白AA-晶状体蛋白和AB-晶状蛋白具有巨大的潜力，因为治疗蛋白对​​当今诊所中一些最常见的眼科疾病的治疗蛋白具有良好的潜力。像SHSP家族的其他成员一样，� -Crystallin低聚物显示出类似伴侣的活性，这些活动是由它们防止其他蛋白质聚集的能力所定义的。在眼外，已证明 - 晶状体可以与在多种疾病的发病机理中共同定位，包括与神经退行性和自身免疫性疾病相关的纤维束。已经证明，通过质粒或病毒表达载体引入SHSP可以在各种疾病模型中挽救表型标记，包括色素性视网膜炎（RP）和易受聚集的性白内障突变体。 与重点在此挑战公告中强调转化科学相一致，我们建议开发基于SHSP的理论，以预防和/或逆转白内障和RP发病机理中实现的聚集体的形成。我们的总体假设是，开发一种将SHSP递送到眼睛的有效方法将为治疗由遗传疾病和代谢压力引起的广泛眼病的新策略提供新的策略。我们的建议围绕两个主要目标组织。在第一个目标中，我们将开发和优化各种技术，以准备装有人类SHSP的纳米颗粒。在第二个目标中，我们将测试通过抑制和/或反转由与RP和白内障相关的突变蛋白形成的高分子量聚集体时通过纳米颗粒传递的SHSP的有效性。 特定目标1：开发一系列SHSP纳米医学公式，并优化以持续缓慢释放到眼睛中。我们将追求四种不同的技术，以使SHSP复合物缓慢释放到眼睛中。源自人类SHSP基因的野生型和“活化”的SHSP亚基的组合将掺入纳米颗粒中。将进行研究以优化不同纳米颗粒配方中的蛋白质释放和组织穿透。 具体目标2：检验SHSP可以抑制由易溶的高分子寡聚复合物和易于聚集的易启动的视紫红质和透镜晶体晶体突变体的不溶性高分子量复合物的形成的假设。在蛋白质聚集疾病的动物模型中，将测试证明在RP和白内障组织培养模型中有效性的纳米颗粒公式，包括RP的p23h转基因大鼠模型和AA-晶体的aa-crystallin敲除白内障的小鼠模型。 公共卫生相关性：该项目旨在开发新药，以预防或逆转盲目疾病，例如白内障和色素性视网膜炎，这些疾病与蛋白质的聚集有关。蛋白质聚集疾病解释了眼睛中一些最常见的退化性疾病。我们提出的研究可以通过防止视力丧失和相关的金融燃烧，因为生产力失去和医疗费用增加，可以大大改善受影响患者的生活质量。