An Experimental Approach to Maculopathy

项目摘要 建议继续研究萎缩性老年性黄斑变性(AMD)的基本机制 通过翻译这部作品来建立新的治疗方法。早期AMD进展为晚期致盲形式的 遵循两种不同途径之一的疾病；萎缩性AMD或地理萎缩(GA)相关 视网膜色素上皮(RPE)进行性衰老和死亡，而脉络膜 新生血管(CNV)与视网膜下新血管的生长有关。在上一次授权期内 我们的假设得到了强有力的支持，即骨形态发生蛋白-4的表达增加 RPE中的(Bmp4)诱导萎缩性AMD的特征，并抑制CNV；从而介导 “分子开关”，它决定了患者患上哪种晚期AMD。对此的主要回应之一 RPE中BMP4表达增加是伴侣蛋白AB晶状体蛋白表达上调。我们 假设在早期AMD中，BMP4表达增加导致RPE衰老和一种新的反应性 RPE中AB晶状体蛋白表达增加所致的神经保护反应 进展，这种反应是不堪重负的，进一步持续的氧化应激导致进展到GA。 然后我们假设AB晶体蛋白衍生的具有伴侣活性的寡肽可以提供类似的 保护RPE免受氧化应激和其他损伤，并可选择性地输送到RPE中。 此外，还可以组装和优化新型AB晶体蛋白多肽纳米颗粒，以有效拯救 培养中的RPE功能障碍，以及具有GA特征的多个小鼠模型。这项工作具有很高的 意义重大，因为目前还没有有效的治疗GA的方法。这些假设将通过使用 以下具体目标：目标1.确定分子、细胞和功能之间的相互关系 RPE中BMP4和AB晶状体蛋白之间的关系以及氧化应激对这些过程的影响。目标2. 开发和优化AB晶体蛋白多肽纳米粒以抑制各种形式的应激对RPE的影响 细胞在培养中生长。目的#3.测定AB晶体蛋白多肽纳米粒在体内的药代动力学 眼内或全身给药，并确定这些纳米粒抑制肿瘤进展的能力 具有GA特征的多种小鼠模型的疾病。

Project Abstract Continued studies on basic mechanisms of atrophic age related macular degeneration (AMD) are proposed with translation of this work to establish novel therapies. Early AMD progresses to late blinding forms of the disease by following one of two divergent pathways; atrophic AMD or geographic atrophy (GA) is associated with progressive senescence and death of the retinal pigment epithelium (RPE), while choroidal neovascularization (CNV) is associated with growth of new vessels under the retina. In the last grant period we developed strong support for our hypothesis that increased expression of bone morphogenetic protein-4 (BMP4) in RPE induces features characteristic of atrophic AMD, and inhibits CNV; thus mediating a "molecular switch" that determines which late form of AMD a patient develops. One of the major responses to increased BMP4 expression in RPE is upregulated protein expression of the chaperone aB crystallin. We hypothesize that in early AMD, increased BMP4 expression leads to RPE senescence and a novel reactive neuroprotective response with increased expression of aB crystallin from RPE; however as disease progresses, this response is overwhelmed and further sustained oxidative stress results in progression to GA. We then hypothesize that aB crystallin-derived oligopeptides with chaperone activity can provide similar protection of RPE from oxidative stress and other injuries and can be selectively transported into the RPE. Furthermore, novel aB crystallin peptide nanoparticles can be assembled and optimized to effectively rescue dysfunctional RPE in culture, and in multiple murine models with features of GA. This work is highly significant since there is currently no effective therapy for GA. The hypotheses will be tested using the following Specific Aims: Aim #1. Determine the molecular, cellular, and functional inter-relationships between BMP4 and aB crystallin in RPE and the effect of oxidative stress on these processes. Aim # 2. Develop and optimize aB crystallin peptide nanoparticles that inhibit effects of various forms of stress in RPE cells grown in culture. Aim # 3. Determine the pharmacokinetics of aB crystallin peptide nanoparticles after intraocular or systemic delivery and determine the ability of these nanoparticles to inhibit progression of disease in multiple murine models with features of GA .