The Neuroprotective Effect of HSP72 Induction in Experimental Glaucoma

摘要 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 青光眼的降眼压治疗现状。我们的假设是长期的药物诱导 HSP72(热休克蛋白70的诱导形式)的表达可保护视网膜节细胞免受青光眼的影响 损坏。这一假说是基于HSP72广泛的细胞保护能力，这可能是 可以用它抑制几种细胞凋亡的能力来解释。 该提案的具体目标是： 目的1.研究热休克蛋白72过表达对大鼠青光眼模型视网膜神经节细胞存活的影响。腺体- 携带HSP72基因的相关病毒(AAV，血清型5)载体(AAV-HSP72)将用于 在视网膜节细胞中传递和表达相应的蛋白，这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抑制实验性细胞凋亡的机制 青光眼。已知HSP72可抑制多种导致细胞死亡的通路及其抗凋亡功能 与细胞凋亡诱导因子(AIF)、APAF-1的结合有关 和c-Jun氨基末端激酶(JNK)。上述机制及研资局其他潜在机制的参与 HSP72对实验性青光眼的保护作用将被研究。这些研究的结果将帮助我们确定 HSP72的神经保护机制和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。我们会 研究GGA和Arimoclomol，HSP72共同诱导剂，作为潜在的治疗药物保护RGCs。 大鼠青光眼模型。HSP72在GGA和Arimoclomol介导的神经保护中的作用将通过 用小干扰RNA(SiRNA)抑制HSP72的表达。 本研究旨在综合评价热休克蛋白72的神经保护作用。 青光眼和了解其对细胞凋亡途径的作用机制。我们相信， 热休克蛋白72的细胞保护和抗凋亡特性及其药物诱导的可能性 这种蛋白在感受压力的细胞中的表达使其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 在青光眼中设计保存视网膜节细胞的策略的因素，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的，这些分子机制具有共同的特征视神经最终通路 损坏和视觉损失。

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.