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: 8523883
• 负责人: NATIK PIRI
• 金额: $ 24.33万
• 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/8523883
• 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; Ganglion Cell Layer; Genes; Glaucoma; Glutamates; HSP72 protein; Heat-Shock Proteins 70; Homeostasis; Hybrids; Hyperthermia; Immunoblotting; Immunohistochemistry; Immunologics; Immunoprecipitation; 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; 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; intravitreal injection; neuronal cell body; neuroprotection; neurotrophic factor; overexpression; promoter; protective effect; protein expression; protein protein interaction; screening; stress-activated protein kinase 1; therapeutic target; vector; yeast two hybrid system

ABSTRACT 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.

摘要 我们研究的长期目标是开发一种神经保护策略， 目前治疗青光眼的IOP降低。我们的假设是长期的药物诱导 热休克蛋白70（Hsp 72）的诱导型表达可保护视网膜节细胞免受肿瘤的侵袭 损害这一假设是基于HSP 72的广泛的细胞保护能力，这可能是由于HSP 72的细胞保护能力的不同而导致的。 这可以通过其抑制几种凋亡途径的能力来解释。 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。腺- 携带HSP 72基因的基于相关病毒（AAV，血清型5）的载体（AAV-HSP 72）将用于 在RGCs中递送和表达相应的蛋白质，这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2. HSP 72抑制实验性肝癌细胞凋亡途径的机制 青光眼已知HSP 72抑制导致细胞死亡的多种途径及其抗凋亡功能 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）。这些和其他潜在的RGC机制的参与 研究HSP 72对实验性青光眼的保护作用。这些研究的结果将帮助我们定义 HSP 72神经保护机制并更好地了解青光眼中的RGC细胞凋亡。 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RGC的潜在治疗剂， 大鼠青光眼模型。将通过以下方法分析HSP 72在GGA和arimoclomol介导的神经保护中的作用： 用设计用于降解HSP 72 mRNA的小干扰RNA（siRNA）抑制HSP 72表达。 本研究旨在全面评价Hsp 72在实验性脑缺血中的神经保护作用。 青光眼，并了解其对细胞凋亡途径的作用机制。我们认为 HSP 72的细胞保护和抗凋亡特性以及HSP 72诱导细胞凋亡的可能性 这种蛋白质在经历应激的细胞中的表达使这种蛋白质成为一种有吸引力的治疗靶点， 青光眼神经保护HSP 72保护细胞免受各种应激刺激的能力是一个重要的因素。 在设计策略以保护青光眼中的RGC时，这是一个重要的因素，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，这些机制具有共同的特征性视神经最终通路， 损伤和视力丧失。

项目成果

Bis(zinc-dipicolylamine), Zn-DPA, a new marker for apoptosis.

• DOI: 10.1167/iovs.13-13346
• 发表时间: 2014-07
• 期刊: Investigative ophthalmology & visual science
• 影响因子: 4.4
• 作者: Jacky M K Kwong;Celia Hoang;Reshil T Dukes;R. Yee;B. Gray;K. Pak;J. Caprioli