The Neuroprotective Effect of HSP72 Induction in Experimental Glaucoma

HSP72 诱导对实验性青光眼的神经保护作用

• 批准号: 7942827
• 负责人: NATIK PIRI
• 金额: $ 26.68万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 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 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7942827
• 关键词: Affect; Affinity; Animals; Apoptosis; Apoptotic; Axon; Binding; Blindness; Blood flow; Cell Death; Cell Survival; Cells; Characteristics; Chickens; Chronic; Complement; Cytomegalovirus; Cytoprotection; Disease; Disease Progression; Early Promoters; Electroporation; Enhancers; Evaluation; Fever; Ganglion Cell Layer; Genes; Glaucoma; Glutamates; HSP72 protein; Heat-Shock Proteins 70; Homeostasis; Hybrids; Immunoblotting; Immunohistochemistry; Immunologics; Immunoprecipitation; Injection of therapeutic agent; Label; Long-Term Effects; Mediating; Messenger RNA; Modeling; Molecular; Neurons; Neuropathy; Neuroprotective Agents; Nitric Oxide; North Carolina; Optic Nerve; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Physiologic Intraocular Pressure; Plasmids; Process; Production; Protein Array; Proteins; Rattus; Reporting; Research; Retina; Retinal; Retinal Ganglion Cells; Risk Factors; Role; Screening procedure; Serotyping; Small Interfering RNA; Sodium Dodecyl Sulfate-PAGE; Stimulus; Stress; Techniques; Therapeutic; Therapeutic Agents; Transfection; Universities; Vaccination; Viral; Visual Fields; Yeasts; Zinc; apoptotic protease-activating factor 1; arimoclomol; base; beta Actin; cell type; design; excitotoxicity; experience; expression vector; gene therapy; geranylgeranylacetone; improved; in vivo; neuronal cell body; neuroprotection; neurotrophic factor; overexpression; promoter; protective effect; protein expression; protein protein interaction; public health relevance; stress-activated protein kinase 1; therapeutic target; vector; yeast two hybrid system

DESCRIPTION (provided by applicant): The long-term objective of our research is to develop a neuroprotective strategy that will complement the current treatment of glaucoma with IOP reduction. Our hypothesis is that long-term pharmacological induction of Hsp72 (the inducible form of heat shock protein 70) expression can protect RGCs from glaucomatous damage. This hypothesis is based on the wide range of cytoprotective capacities of HSP72, which may be explained by its ability to inhibit several apoptotic pathways. Specific aims of the proposal are: Aim 1. Determine the effect of Hsp72 overexpression on RGC survival in a rat glaucoma model. Adeno- associated viral (AAV, serotype 5) based vectors carrying the HSP72 gene (AAV-HSP72) will be used to deliver and express the corresponding protein in RGCs, which is the primary target for neuroprotection. Specific overexpression of HSP72 in glaucomatous retinas will allow us to evaluate its neuroprotective effect. Aim 2. Elucidate the mechanisms of inhibition of apoptotic pathways by HSP72 in experimental glaucoma. HSP72 is known to inhibit multiple pathways leading to cell death and its anti-apoptotic functions are associated with binding to apoptosis protease activating factor-1 (Apaf-1), apotosis-inducing factor (AIF) and c-Jun N-terminal kinase (JNK). The involvement of these and other potential mechanisms of RGC protection by HSP72 in experimental glaucoma will be studied. The results of these studies will help us define HSP72 neuroprotective mechanisms and better understand RGC apoptosis in glaucoma. Aim 3. Evaluate the therapeutic potential of pharmacological induction of HSP72 for glaucoma. We will study GGA and arimoclomol, HSP72 co-inducers, as potential therapeutic agents for protection of RGCs in the rat glaucoma model. The role of HSP72 in GGA and arimoclomol mediated neuroprotection will be analyzed by inhibition of HSP72 expression with small interfering RNA (siRNA) designed to degrade HSP72 mRNA. The proposal is intended to comprehensively evaluate the Hsp72 neuroprotective effect in experimental glaucoma and understand the mechanisms of its action on apoptotic pathways. We believe that the cytoprotective and anti-apoptotic characteristics of HSP72 and the possibility to pharmacologically induce expression of this protein in cells experiencing stress make this protein an attractive therapeutic target for glaucoma neuroprotection. The ability of HSP72 to protect cells from a variety of stress stimuli is an important factor in designing a strategy to preserve RGCs in glaucoma, since the cellular damage in this disease may be caused by different molecular mechanisms that have a common final pathway of characteristic optic nerve damage and visual loss. PUBLIC HEALTH RELEVANCE: The degeneration of retinal ganglion cells (RGCs) is the primary cause of visual loss in glaucoma, which affects more than 70 million people worldwide. The main aim of the proposed study is the evaluation of the heat shock protein 72 (HSP72) induction with non-toxic drugs in RGC protection in animals with glaucoma. This protein is known to protect different types of cells, including neuronal cells, against various insults. The results of our studies may provide support for therapeutic consideration of HSP72 inducers as potential neuroprotective drugs for glaucoma.

描述（由申请人提供）：我们研究的长期目标是开发一种神经保护策略，以补充目前通过降低眼压来治疗青光眼的方法。我们的假设是，长期药理学诱导 Hsp72（热休克蛋白 70 的诱导形式）表达可以保护 RGC 免受青光眼损伤。该假说基于 HSP72 广泛的细胞保护能力，这可以通过其抑制多种细胞凋亡途径的能力来解释。该提案的具体目标是： 目标 1. 确定 Hsp72 过表达对大鼠青光眼模型中 RGC 存活的影响。携带 HSP72 基因 (AAV-HSP72) 的腺相关病毒（AAV，血清型 5）载体将用于在 RGC 中递送和表达相应的蛋白质，这是神经保护的主要目标。 HSP72 在青光眼视网膜中的特异性过度表达将使我们能够评估其神经保护作用。目标 2. 阐明 HSP72 在实验性青光眼中抑制细胞凋亡途径的机制。 HSP72 已知可抑制导致细胞死亡的多种途径，其抗凋亡功能与与凋亡蛋白酶激活因子 1 (Apaf-1)、凋亡诱导因子 (AIF) 和 c-Jun N 末端激酶 (JNK) 的结合有关。将研究实验性青光眼中 HSP72 保护 RGC 的这些机制和其他潜在机制。这些研究的结果将帮助我们定义 HSP72 的神经保护机制，并更好地了解青光眼中 RGC 的凋亡。目标 3. 评估药理诱导 HSP72 对青光眼的治疗潜力。我们将研究 GGA 和阿莫洛莫（HSP72 共诱导剂）作为保护大鼠青光眼模型中 RGC 的潜在治疗剂。 HSP72 在 GGA 和阿莫洛莫介导的神经保护中的作用将通过用设计用于降解 HSP72 mRNA 的小干扰 RNA (siRNA) 抑制 HSP72 表达来分析。该提案旨在全面评估Hsp72对实验性青光眼的神经保护作用，并了解其对细胞凋亡通路的作用机制。我们相信，HSP72 的细胞保护和抗凋亡特性以及在经历应激的细胞中药理学诱导该蛋白表达的可能性使得该蛋白成为青光眼神经保护的有吸引力的治疗靶点。 HSP72 保护细胞免受各种应激刺激的能力是设计保护青光眼 RGC 策略的重要因素，因为这种疾病中的细胞损伤可能是由不同的分子机制引起的，这些分子机制具有特征性视神经损伤和视力丧失的共同最终途径。公众健康相关性：视网膜神经节细胞 (RGC) 变性是青光眼导致视力丧失的主要原因，全世界有超过 7000 万人受到青光眼的影响。本研究的主要目的是评估无毒药物诱导热休克蛋白 72 (HSP72) 对青光眼动物 RGC 的保护作用。众所周知，这种蛋白质可以保护不同类型的细胞（包括神经元细胞）免受各种损伤。我们的研究结果可能为 HSP72 诱导剂作为青光眼潜在神经保护药物的治疗考虑提供支持。