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

基本信息

批准号：
8323418
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
项目类别：
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/8323418
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 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 intravitreal injection 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

项目摘要

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.

抽象的我们研究的长期目标是开发一种神经保护策略，以补充目前青光眼的治疗方法是降低眼压。我们的假设是长期药理诱导 Hsp72（热休克蛋白 70 的诱导形式）表达可以保护 RGC 免受青光眼的影响损害。该假设基于 HSP72 广泛的细胞保护能力，这可能是这是因为它能够抑制多种细胞凋亡途径。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。腺苷- 携带 HSP72 基因 (AAV-HSP72) 的相关病毒（AAV，血清型 5）载体将用于在 RGC 中递送并表达相应的蛋白质，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。目的2. 阐明HSP72在实验中抑制细胞凋亡途径的机制青光眼。 HSP72 已知可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) 的结合有关和 c-Jun N 末端激酶 (JNK)。 RGC 的这些机制和其他潜在机制的参与将研究 HSP72 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 HSP72 神经保护机制并更好地了解青光眼中 RGC 凋亡。目标 3. 评估药理学诱导 HSP72 对青光眼的治疗潜力。我们将研究 GGA 和 arimoclomol（HSP72 共诱导剂）作为保护 RGC 的潜在治疗剂大鼠青光眼模型。 HSP72 在 GGA 和阿莫洛莫介导的神经保护中的作用将通过使用旨在降解 HSP72 mRNA 的小干扰 RNA (siRNA) 抑制 HSP72 表达。该提案旨在在实验中全面评价Hsp72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。我们相信 HSP72 的细胞保护和抗凋亡特性以及药理学诱导的可能性这种蛋白质在经历应激的细胞中表达使这种蛋白质成为有吸引力的治疗靶点青光眼神经保护。 HSP72 保护细胞免受各种应激刺激的能力是一个重要的设计保护青光眼 RGC 策略的因素，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，但具有共同的特征性视神经最终通路损伤和视力丧失。