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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

基本信息

批准号：
7945316
负责人：
Jonathan Mark Petrash
金额：
$ 49.32万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
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/7945316
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

项目摘要

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(由申请人提供)：本申请涉及广泛的挑战领域(15)转化科学和具体的挑战主题，15-EY-101：眼部退行性疾病中的蛋白质错误折叠。确定可防止蛋白质错误折叠/聚集的治疗性药理制剂/药物可为治疗这些疾病提供新的工具。虽然眼内细胞的退化是各种疾病的基础，包括白内障、老年性黄斑变性和青光眼，但目前还没有被批准的治疗药物来治疗与这些疾病相关的细胞退化。开发这种治疗剂的一个困难是这些退行性疾病的复杂病因，其中多个细胞路径被改变。因此，一种理想的治疗候选药物将针对细胞退化的多种应激途径。我们的项目解决了基于蛋白质的眼部治疗的一个关键难题：如何在为长期治疗提供治疗性蛋白质的同时，将重复眼内注射的风险和成本降至最低。用于治疗眼部退行性疾病的药物很可能从确诊之日起就开始使用。目前批准的眼部蛋白质疗法的半衰期相对较短(10天)，这需要大约每6周向眼睛重复注射一次。因此，需要缓释系统来延长大分子向眼睛的输送。将大分子定向输送到眼睛退化细胞的技术也是一个重要的目标。蛋白质药物除了对普遍存在的蛋白水解酶敏感外，由于其大分子尺寸，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。小分子热休克蛋白(SHSP)蛋白AA-晶状体蛋白和AB-晶状体蛋白作为治疗当今临床上最常见的眼病的蛋白质具有很好的潜力。像SHSP家族的其他成员一样，晶体蛋白寡聚体表现出类似伴侣的活性，这取决于它们阻止其他蛋白质聚集的能力。在眼睛之外，晶体蛋白已被证明与多种疾病的发病机制中涉及的蛋白质聚集体共定位，包括与神经退行性疾病和自身免疫性疾病相关的纤维束。通过质粒或病毒表达载体导入sHSPs已被证明在各种疾病模型中拯救表型标记，包括视网膜色素变性(RP)和易聚集的白内障突变体。与本挑战公告中对翻译科学的强调一致，我们建议开发基于SHSP的治疗方法，以防止和/或逆转与白内障和RP发病机制有关的聚集体的形成。我们的主要假设是，开发一种有效的将sHSPs输送到眼睛的方法将为治疗由遗传疾病和代谢应激引起的广泛眼部疾病提供一种新的策略。我们的提案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制备载人SHSP纳米颗粒的技术。在第二个目标中，我们将测试通过纳米颗粒传递的SHSP在抑制和/或逆转与RP和白内障相关的突变蛋白形成的高分子量聚集体方面的有效性。具体目标1：开发一系列SHSP纳米药物配方，并优化其在眼内的持续缓释。我们将寻求四种不同的技术来产生SHSP复合体进入眼睛的缓慢释放。来自人类SHSP基因的野生型和“激活的”SHSP亚基的组合将被整合到纳米颗粒中。将开展研究，以优化不同纳米颗粒配方的蛋白质释放和组织渗透。特定目的2：验证SHSP可以抑制由易于聚集的视紫红质和晶状体晶体蛋白突变体启动的不溶性高分子量低聚复合体的形成的假设。在RP和白内障的组织培养模型中证明有效的纳米颗粒配方将在蛋白质聚集性疾病的动物模型中进行测试，包括P23H转基因RP大鼠模型和AA-晶体蛋白敲除小鼠白内障模型。公共卫生相关性：该项目寻求开发新的药物来预防或逆转与蛋白质聚集有关的白内障和视网膜色素变性等致盲疾病。蛋白质聚集性疾病是眼部最常见的退行性疾病之一。我们建议的研究可以通过预防由于生产力下降和医疗费用增加而造成的视力丧失和相关的经济负担，显著提高受影响患者的生活质量。