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: 7735983
• 负责人: NATIK PIRI
• 金额: $ 26.95万
• 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/7735983
• 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(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，以补充目前青光眼治疗的降低眼压。我们的假设是，长期药物诱导HSP72(热休克蛋白70的诱导形式)的表达可以保护视网膜节细胞免受青光眼的损害。这一假说是基于HSP72广泛的细胞保护能力，这可能是因为它能够抑制几种凋亡途径。该提案的具体目的是：目的1.确定HSP72过表达对大鼠青光眼模型RGC存活的影响。基于携带HSP72基因的腺相关病毒(AAV，血清型5)的载体(AAV-HSP72)将被用来在RGC中传递和表达相应的蛋白，RGC是神经保护的主要靶点。HSP72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。目的2.阐明热休克蛋白72抑制实验性青光眼细胞凋亡的机制。HSP72可抑制多种途径导致细胞死亡，其抗凋亡作用与与细胞凋亡相关的蛋白水解酶激活因子-1(APAF-1)、细胞凋亡诱导因子(AIF)和c-jun氨基末端激酶(JNK)的结合有关。这些和HSP72保护RGC的其他潜在机制在实验性青光眼中的参与将被研究。这些研究的结果将有助于我们确定HSP72的神经保护机制，并更好地了解青光眼中RGC的凋亡。目的3.评价HSP72药物诱导治疗青光眼的可能性。我们将研究GGA和Arimoclomol，HSP72的联合诱导剂，作为潜在的治疗药物来保护大鼠青光眼模型的视网膜节细胞。HSP72在GGA和Arimoclomol介导的神经保护中的作用将通过用旨在降解HSP72 mRNA的小干扰RNA(SiRNA)来抑制HSP72的表达来分析。该建议旨在全面评价HSP72对实验性青光眼的神经保护作用，并了解其对细胞凋亡途径的作用机制。我们认为，HSP72的细胞保护和抗凋亡特性，以及在经历应激的细胞中药物诱导该蛋白表达的可能性，使其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青光眼视网膜神经节细胞保护策略的重要因素，因为在青光眼中，细胞损伤可能是由不同的分子机制引起的，这些分子机制具有共同的最终通路，即特征性视神经损伤和视力丧失。公共卫生相关性：视网膜神经节细胞(RGC)的退化是青光眼视力丧失的主要原因，全球有超过7000万人受到青光眼的影响。本研究的主要目的是评价无毒药物诱导的热休克蛋白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，包括神经细胞，免受各种侮辱。我们的研究结果可能为HSP72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。