Acid-sensing channels as novel target for brain ischemia

描述（由申请人提供）：缺血性卒中是死亡率和发病率的主要原因，也是长期残疾的最常见原因。然而，除了使用溶栓药物外，目前还没有有效的治疗方法，这些药物的治疗时间窗有限，并且可能有颅内出血的副作用。几十年来，人们已经认识到细胞内Ca 2+积累，特别是通过谷氨酸受体激活，以及由此产生的Ca 2+毒性，在缺血性脑损伤中起重要作用。然而，最近的临床试验使用谷氨酸拮抗剂和最近的自由基清除剂，未能显示对缺血性损伤的保护。尽管多种因素可能共同导致试验失败，但谷氨酸非依赖性Ca2+负荷途径可能同样导致缺血中的Ca2+毒性。事实上，我们最近在神经元细胞培养和整个缺血动物模型中的研究已经证明，酸敏感离子通道（ASIC）的激活和随后的Ca2+进入在很大程度上是酸中毒介导的、谷氨酸受体非依赖性缺血性脑损伤的原因。在培养的小鼠皮层神经元中，降低pH激活阿米洛利敏感的ASIC电流。在大多数这些神经元中，ASIC对Ca 2+是可渗透的，并且这些通道的激活诱导细胞内Ca 2+（[Ca 2 +]i）的浓度增加。在电压门控Ca 2+通道和谷氨酸受体阻滞剂存在的情况下，通过将神经元与酸性溶液短暂孵育来激活ASIC会诱导时间依赖性细胞损伤。然而，这种酸诱导的谷氨酸非依赖性神经元损伤通过阻断ASIC、通过减少细胞外[Ca2+]或通过ASIC1基因敲除来抑制。在体内小鼠缺血模型中，ASIC1阻断或ASIC1基因敲除显著减少梗死体积。这些发现有力地表明，ASIC可能是人类卒中的新治疗靶点。我们的目标是将我们在动物细胞中的令人兴奋的发现扩展到人脑神经元，以探索ASIC在酸中毒介导的人脑神经元缺血性损伤中的作用。我们的中心假设是人脑神经元表达ASIC。ASICs的激活诱导细胞内Ca2+积累，这参与酸中毒介导的谷氨酸非依赖性神经元损伤。这些研究的成功是建立ASIC作为人类卒中治疗新靶点的重要一步。公共卫生相关性：我们计划研究预防中风后细胞丢失的新机制和策略。我们以前的研究结果强烈表明，ASIC可能是人类卒中的新治疗靶点。这些研究的成功是建立ASIC作为人类卒中治疗新靶点的重要一步。

DESCRIPTION (provided by applicant): Ischemic stroke is a leading course of mortality and morbidity, and the most common reason for long-term disabilities. Unfortunately there is still no effective treatment for stroke patients other than the use of thrombolitics which have limited time window and potential side effect of intracranial hemorrhage. It has been well-recognized for several decades that that intracellular Ca2+ accumulation, particularly through glutamate receptor activation, and the resultant Ca2+ toxicity, play an important role in ischemic brain injury. However, the recent clinical trials using glutamate antagonists and most recently free radical scavengers, have failed to show protection against ischemic injury. Although multiple factors may have contributed together to the failure of the trials, it is likely that glutamate-independent Ca2+ loading pathway(s) equally contribute to the Ca2+ toxicity in ischemia. Indeed, our recent studies in neuronal cell culture and in whole animal models of ischemia have demonstrated that activation of acid-sensing ion channels (ASICs), and subsequent Ca2+ entry are largely responsible for acidosis- mediated, glutamate receptor-independent ischemic brain injury. In cultured mouse cortical neurons, lowering pH activates amiloride-sensitive ASIC currents. In the majority of these neurons, ASICs are permeable to Ca2+, and activation of these channels induces increased concentration of intracellular Ca2+ ([Ca2+]i). Activation of ASICs by brief incubation of neurons with acidic solutions induces time-dependent cell injury in the presence of blockers for voltage-gated Ca2+ channels and the glutamate receptors. This acid-induced, glutamate-independent neuronal injury is, however, inhibited by blocking the ASICs, by reducing the extracellular [Ca2+], or by ASIC1 gene knockout. In in vivo mouse model of ischemia, ASIC1 blockade or ASIC1 gene knockout dramatically reduced infarct volume. These findings strongly suggest that ASICs may represent novel therapeutic targets for human stroke. Our objective is to extend our exciting findings in animal cells to human brain neurons to explore the role of ASICs in acidosis-mediated ischemic injury of human brain neurons. Our central hypothesis is that human brain neurons express ASICs. Activation of ASICs induces intracellular Ca2+ accumulation, which is involved in acidosis-mediated, glutamate-independent neuronal injury. Success of these studies is an important step for establishing ASICs as novel targets for human stroke therapy. PUBLIC HEALTH RELEVANCE: We plan to investigate novel mechanisms and strategies to prevent cell loss after stroke. Our previous findings strongly suggest that ASICs may represent novel therapeutic targets for human stroke. Success of these studies is an important step for establishing ASICs as novel targets for human stroke therapy.