An exploratory proposal to move select Drosophila nociception screen hits into mouse models

项目总结/摘要 疼痛和疼痛致敏是组织损伤、癌症治疗和某些疾病的毁灭性副作用 比如糖尿病有效治疗疼痛是许多损伤和疾病状态的巨大临床障碍。 疼痛研究的领导者认为，新的模型，其中新的保守基因靶可以有效地 已确定迫切需要。更重要的是新的保守目标功能的翻译 在这些新的和简单的模型中识别出更多临床相关的脊椎动物模型。我们的长期目标是 以鉴定具有高度临床转化潜力的新型疼痛/疼痛致敏调节剂。过去十 多年来，我的实验室在果蝇中进行了大量的基因筛选， 独特的组织损伤和疼痛检测这些筛选已经确定了高度保守的疼痛调节因子 生物学（特别是组织损伤后的急性和慢性疼痛致敏），其在调节 以前在其他模型中的工作中没有发现疼痛。两个最有前途的屏幕点击， 定义为果蝇表型的保守性、稳健性和独特性，以及对疼痛的新奇。 在生物学上，Smoothened是Hh信号通路的信号转导子，胰岛素受体。飞 幼虫，这两个基因都需要在外周伤害性感觉神经元-平滑，以调节急性 热痛致敏和胰岛素受体（InR）调节急性致敏的停止。InR是 特别有趣的是，神经元特异性的作用，调节从急性到慢性敏化的过渡 可能会提供一种全新的方式来看待与糖尿病相关的疼痛。我们的目标是探索 R21提案是必要和紧迫的。对于Smoothened和胰岛素受体，我们建议执行 在小鼠中进行的平行实验与我们已经在苍蝇中进行的实验相同：我们将有条件地敲 这些基因在疼痛感觉神经元中表达，并评估基线，急性和慢性疼痛表型。 两次标准测定。这是必要的，因为这种基于果蝇的结果的正向翻译， 尽管在发育生物学和先天免疫中是标准的和有用的， 在疼痛领域的任何系统的方式。这是紧迫的，因为成功的翻译（意思是基因做 实际上以某种方式调节小鼠的疼痛生物学）可以建立一个真正的新的保守疼痛管道 用于脊椎动物模型测试的基因靶点。尤其是我们还有许多其他保守的靶点 在我们的屏幕上已经被识别出来了。这些结果可能是科学上令人信服的-提供新的 对疼痛反应的保守调节的见解。它们也可能在临床上有用--为我们指明了方向， 新的可药用保守靶点，在其他互补靶点中难以系统地鉴定， 实验系统疼痛感知和敏感机制对动物的重要性，反映在 这些机制的高度进化保守性，向我们表明这种方法可能有效。

PROJECT SUMMARY/ABSTRACT Pain and pain sensitization are devastating side effects of tissue injury, cancer treatment, and certain diseases like diabetes. Treating pain effectively is an enormous clinical obstacle for many injuries and disease states. Pain research leaders have argued that new models in which novel conserved gene targets can be efficiently identified are urgently needed. Even more essential is the translation of new conserved targets functionally identified in these new and simple models to more clinically relevant vertebrate models. Our long-term goal is to identify novel pain/pain sensitization regulators with high potential for clinical translation. Over the last ten years my laboratory has performed a number of genetic screens in the fruit fly Drosophila, by combining unique tissue damage and pain assays. These screens have identified highly conserved regulators of pain biology (in particular acute and chronic pain sensitization following tissue damage) whose roles in regulating pain were not previously appreciated from work in other models. The two most promising screen hits as defined by conservation, robustness and uniqueness of their Drosophila phenotype, and novelty to pain biology, are Smoothened, the signal transducer of the Hh signaling pathway, and the Insulin receptor. In fly larvae, both genes are required in peripheral nociceptive sensory neurons– Smoothened to regulate acute thermal pain sensitization and the Insulin receptor (InR) to regulate the cessation of acute sensitization. InR is particularly interesting as the neuron-specific role in regulating the transition from acute to chronic sensitization may provide an entirely new way of looking at the pain associated with diabetes. Our goal in this exploratory R21 proposal is necessary and urgent. For both Smoothened and the Insulin Receptor we propose to perform parallel experiment(s) in mice to those that we have already performed in flies: we will conditionally knock these genes out in pain-sensing sensory neurons and assess baseline, acute, and chronic pain phenotypes in two standard assays. This is necessary because this type of forward translation of Drosophila-based results, though standard and useful in developmental biology and innate immunity, has not been previously attempted in any systematic way in the pain field. It is urgent because successful translation (meaning the genes do actually regulate pain biology in some way in mice) could establish a veritable pipeline of new conserved pain gene targets for testing in vertebrate models. This is especially true as we have many other conserved targets that have been identified in our screens. The results could be both scientifically compelling- providing new insight into conserved regulation of pain responses. They could also be clinically useful- pointing the way to new druggable conserved targets that have been difficult to identify systematically in other complementary experimental systems. The importance of pain-sensing and sensitizing mechanisms to the animal, reflected in the high evolutionary conservation of these mechanisms, suggest to us that this approach is likely to work.