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: 8128496
• 负责人: NATIK PIRI
• 金额: $ 25.61万
• 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/8128496
• 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; Health; Heat-Shock Proteins 70; Homeostasis; Hybrids; Immunoblotting; Immunohistochemistry; Immunologics; Immunoprecipitation; Injection of therapeutic agent; Label; Long-Term Effects; Mediating; Messenger RNA; Modeling; Molecular; N-terminal; Neurons; Neuropathy; Neuroprotective Agents; Nitric Oxide; North Carolina; Optic Nerve; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Phosphotransferases; 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; Stimulus; Stress; Techniques; Therapeutic; Therapeutic Agents; Transfection; Universities; Vaccination; Viral; Visual Fields; Yeasts; Zinc; 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; 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，该策略将补充目前的青光眼治疗，降低IOP。我们的假设是，长期的药物诱导热休克蛋白72（热休克蛋白70的诱导型）的表达可以保护视网膜神经节细胞免受神经胶质瘤的损害。这一假说是基于热休克蛋白72的广泛的细胞保护能力，这可能是由于它能够抑制几种凋亡途径。该提案的具体目标是：目标1。确定Hsp 72过表达对大鼠青光眼模型中RGC存活的影响。携带HSP 72基因的基于腺相关病毒（AAV，血清型5）的载体（AAV-HSP 72）将用于在RGC中递送和表达相应的蛋白质，其是神经保护的主要靶标。HSP 72在青光眼视网膜中的特异性过表达将使我们能够评估其神经保护作用。目标二。探讨热休克蛋白72抑制青光眼细胞凋亡的机制。已知HSP 72抑制导致细胞死亡的多种途径，并且其抗凋亡功能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HSP 72在实验性青光眼中保护RGC的这些和其他潜在机制的参与。这些研究结果将有助于我们确定HSP 72的神经保护机制，更好地了解青光眼RGC凋亡。目标3。评估药物诱导HSP 72治疗青光眼的潜力。我们将研究GGA和arimoclomol，HSP 72共诱导剂，作为保护大鼠青光眼模型中RGCs的潜在治疗剂。HSP 72在GGA和arimoclomol介导的神经保护中的作用将通过用设计用于降解HSP 72 mRNA的小干扰RNA（siRNA）抑制HSP 72表达来分析。本研究旨在全面评价热休克蛋白72对实验性青光眼的神经保护作用，并了解其对凋亡通路的作用机制。我们认为，HSP 72的细胞保护和抗凋亡特性以及在经历应激的细胞中诱导该蛋白表达的可能性使该蛋白成为青光眼神经保护的有吸引力的治疗靶点。HSP 72保护细胞免受各种应激刺激的能力是设计保护青光眼中RGC的策略的重要因素，因为这种疾病中的细胞损伤可能由不同的分子机制引起，这些分子机制具有共同的特征性视神经损伤和视力丧失的最终途径。公共卫生关系：视网膜神经节细胞（RGC）的变性是青光眼视力丧失的主要原因，全世界有超过7000万人受到影响。本研究的主要目的是评价无毒药物诱导热休克蛋白72（HSP 72）对青光眼动物RGC的保护作用。已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HSP 72诱导剂作为潜在的青光眼神经保护药物的治疗考虑提供支持。