Disease Models for High Throughput Screens

 描述(由申请人提供)：我们的研究是由这样一个前景驱动的，即光感受器退化的进行性本质为延缓细胞损伤和先发制人视力丧失的治疗提供了机会。人类在很大程度上依赖于视锥细胞介导的中央视觉。然而，在相当数量的人中，视杆细胞的死亡或功能障碍会导致视锥细胞的继发性丢失、视网膜回路的重塑和失明。视网膜色素变性(RP)潜在的分子缺陷的异质性使得识别导致变性的具体机制变得困难。更不为人所知的是，不表达突变基因的光感受器继发性死亡的主要原因。迫切需要健壮的模型来概括导致疾病的病理序列，并快速可靠地筛选出减轻疾病影响的因素。我的实验室利用斑马鱼的遗传操作来识别调控光感受器发育和存活的基因。在这项提案中，我们将直接解决在受到广泛光感受器疾病影响的人中观察到的两个共同特征：导致最初光感受器死亡的细胞变化，以及导致失明的继发性变化。我们提出了一个双管齐下的方法：1)应用创新的遗传操作，在斑马鱼视紫红质中引入类似的突变，作为精确定义的人类视网膜疾病模型；2)使用这些和我们现有的退化模型来筛选改变疾病进展的小分子。具体目标的完成将大大促进现场对变更的理解 因此，我们可以研究光感受器死亡的潜在原因，并确定潜在的新靶点或治疗剂作为先导化合物，以保护其免受光感受器退化的次要后果。

 DESCRIPTION (provided by applicant): Our research is driven by the prospect that the progressive nature of photoreceptor degeneration leaves open an opportunity for treatments that delay the time-course of cellular damage and forestall vision loss. Humans are largely dependent upon cone-mediated central vision. However, in a significant number of people, death or dysfunction of rods results in the secondary loss of cones, remodeling of retinal circuitry and blindness. The heterogeneous nature of the molecular defects underlying retinitis pigmentosa (RP) has made identifying the specific mechanisms leading to degeneration difficult. Even less understood, are the principal causes of the secondary death of photoreceptors not expressing the mutated gene. There exists a critical need for robust models that recapitulate the pathological sequence leading to disease and to rapidly and reliably screen for agents to lessen the impact of the disease. My laboratory has taken advantage of the genetic manipulations available in zebrafish to identify genes regulating photoreceptor development and survival. In this proposal, we will directly address two common features observed in people affected by a wide range of photoreceptor disease; the cellular alterations that precipitating initial photoreceptor death, and secondary alterations leading to blindness. We propose a two prong approach: 1) Apply innovative genetic manipulations to introduce analogous mutations in zebrafish rhodopsin as precisely defined models of human retinal disease; 2) Use these and our existing degeneration models to screen for small molecules that modify disease progression. Completion of the specific aims will significantly advance the field's understanding of alterations underlying photoreceptor death, and identify potential novel targets or therapeutic agents as lead compounds to confer protection from the secondary consequences of photoreceptor degeneration.