Crystallins as Neuroprotective and Neuroregenerative Treatments in Experimental Glaucoma

晶状体蛋白作为实验性青光眼的神经保护和神经再生治疗

• 批准号: 263817636
• 负责人: Professorin Dr. Verena Prokosch-Willing
• 金额: --
• 依托单位: Klinik und Poliklinik für allgemeine Augenheilkunde
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/263817636?language=en
• 关键词: Crystallins; Neuroprotective; Neuroregenerative; Treatments; Experimental

Glaucoma is a leading cause of blindness, characterized by progressive retinal ganglion cell (RGC) loss. Lowering of intraocular pressure (IOP), as a key risk factor, is currently the only treatment to slow glaucoma. However, RGCs continue to undergo apoptosis despite IOP lowering . The situation is exacerbated by RGCs, as CNS neurons, lacking the ability to regenerate their axons. A better understanding of the mechanisms and associated with RGC injury, apoptosis and regenerative failure remains an enormous challenge. In our first funding period (PR 1569/1-1) “Molecular biomarkers in the treatment of experimental glaucoma and their functional impact on apoptotic- and regeneration-related signalling pathways”, we analysed different glaucoma models in detail at functional, cellular and molecular levels. We established a proteomic profile, identified molecular markers and looked into neuroprotection and neuroregeneration related to them. Crystallins were found to be one of the most striking markers. We deciphered, that crystallins appeared to show very specific expression patterns in retinal and vitreous samples in the course of the disease. Intravitreal injection of alphaA and betaB2 in vivo exerted neuroprotective effects. BetaB2 crystallin and betaB2 expressing NPCs given intravitreally even promoted neuroregeneration. Besides that crystallins were uptaken from the medium into the cells, transported to the site of lesion and seemed to exert effects via neurotrophic and calcium-dependent pathways. However, crystallins- known as heat-shock proteins- comprise a whole family of alpha (A and B), beta (A1/3, A2, A4, B1, B2, B3) und gamma (A, B, C, D, E, S) crystallins. The importance and interplay of the individual crystallins, the precise neuroprotective and neuroregenerative mechanisms still remain unclear and must be elucidated before their potential in the treatment of glaucoma can be realized. This will be addressed in the second funding period.Firstly, targeted proteomics will help to analyse the whole crystallin family including the less abundant crystallins and their role in regenerative and degenerative pathways. The 5 key crystallins will be identified, corresponding relevant interactions and signalling pathways deciphered. The neuroprotective and regenerative potential of the five presumably most powerful will be assessed by examining retinal explants and isolated RGCs under regenerative and degenerative conditions in vitro. Blocking experiments and specific analysis of the tissue and supernatant will verify the signalling pathways. The last step will analyse neuroprotection and regeneration of the two most promising crystallins in vivo, with the aim of proving novel treatment alternatives for glaucoma.

青光眼是导致失明的主要原因，其特征在于进行性视网膜神经节细胞（RGC）损失。降低眼内压（IOP）作为一个关键的危险因素，是目前唯一的治疗方法来减缓青光眼。然而，尽管IOP降低，RGCs继续进行凋亡。RGC作为CNS神经元，缺乏再生轴突的能力，使情况恶化。更好地了解RGC损伤、凋亡和再生失败的机制仍然是一个巨大的挑战。在我们的第一个资助期（PR 1569/1-1）“实验性青光眼治疗中的分子生物标志物及其对凋亡和再生相关信号通路的功能影响”中，我们在功能，细胞和分子水平上详细分析了不同的青光眼模型。我们建立了一个蛋白质组学图谱，确定了分子标记，并研究了与它们相关的神经保护和神经再生。晶体蛋白被发现是最引人注目的标志之一。我们发现，在疾病过程中，晶状体蛋白似乎在视网膜和玻璃体样本中表现出非常特定的表达模式。体内玻璃体内注射alphaA和betaB 2具有神经保护作用。玻璃体内给予β B2晶体蛋白和表达β B2的NPC甚至促进神经再生。此外，晶体蛋白从培养基中被摄取到细胞中，运输到损伤部位，似乎通过神经营养和钙依赖性途径发挥作用。然而，晶体蛋白-称为热休克蛋白-包括α（A和B）、β（A1/3、A2、A4、B1、B2、B3）和γ（A、B、C、D、E、S）晶体蛋白的整个家族。个体晶体蛋白的重要性和相互作用，精确的神经保护和神经再生机制仍然不清楚，必须在实现其治疗青光眼的潜力之前阐明。第一，有针对性的蛋白质组学将有助于分析整个晶体蛋白家族，包括数量较少的晶体蛋白及其在再生和退化途径中的作用。将确定5种关键晶体蛋白，并破译相应的相关相互作用和信号通路。将通过在体外再生和变性条件下检查视网膜外植体和分离的RGC来评估五种可能最强大的神经保护和再生潜力。组织和上清液的阻断实验和特异性分析将验证信号通路。最后一步将分析两种最有前途的晶体蛋白在体内的神经保护和再生作用，目的是为青光眼提供新的治疗选择。