Thrombin mediated cytotoxicity during cerebral ischemia

我们提出了一个新颖而令人兴奋的假设--基于文献和我们自己的试验数据-- 神经元损伤通过凝血酶及其受体PAR-1刺激星形胶质细胞激活 由此产生的星形胶质细胞对损伤的反应产生了对神经元的可传递旁分泌保护。 神经元、星形胶质细胞、内皮细胞、周细胞和少突胶质细胞共同组成 神经血管单位，或NVU。从几个目标生成的数据将产生对 有充分证据的预适应现象的机制；将提供相当大的 对脑损伤易感性差异的机制的洞察；并可能产生 中风患者的治疗选择。为了实现这些目标，我们正在创造新的条件 基因敲除小鼠将为更广泛的研究领域提供研究工具。上一次 奖项，R01 NS075930，涵盖2011年8月15日至2015年3月31日，涉及4个AIMS。所有建议 工作已经完成并出版或正在审查中。我们之前获奖的数据显示 已经以两项资助的临床试验的形式对中风护理产生了重大影响。然而，我们的 先前的数据不能完全解释NVU各要素之间的已知差异， 具体地说，星形胶质细胞和内皮细胞比神经元更能耐受缺血。是什么 凝血酶介导的预适应的机制，我们是否可以将该机制与 其他形式的预适应和周适应？为什么大脑中含有凝血酶原mRNA和 凝血酶激活大脑中的蛋白质有什么作用？我们能推导出 洞察大脑对损伤的反应，特别是探索细胞亚型 星形胶质细胞、神经元、周细胞、少突胶质细胞和内皮细胞在损伤过程中的相互作用 细胞？为了回答这些问题，并解决本奖项的中心假设 应用程序，我们在我们的曲目中添加了重要的新模型，并生成了试验数据 谈到提出新的具体目标的可行性和新颖性。

We propose a novel and exciting hypothesis—based on literature and our own pilot data—that neuronal injury stimulates astrocyte activation via thrombin and its receptor PAR-1, and the resulting astrocyte response to injury yields transmissible paracrine protection of neurons. Together, neurons, astroctyes, endothelial cells, pericytes, and oligodendroglia comprise the neurovascular unit, or NVU. Data generated from the several aims will yield insights into the mechanism for the well-documented phenomenon of pre-conditioning; will provide considerable insights into the mechanisms of differential vulnerability to cerebral injury; and would likely yield treatment options for stroke patients. To accomplish the aims, we are creating novel conditional knock-out mice that will provide research tools to the broader research field. The previous award, R01 NS075930, covered 8/15/2011 to 03/31/2015 and involved 4 aims. All proposed work has been completed and published or under review. The data from our prior award has already influenced stroke care significantly, in the form of two funded clinical trials. However, our prior data could not fully explain the known differences among the elements of the NVU, specifically, that astrocytes and endothelial cells tolerate ischemia better than neurons. What is the mechanism of thrombin-mediated pre-conditioning, and can we relate that mechanism to other forms of pre- and peri-conditioning? Why does the brain contain prothrombin mRNA and protein and what purpose could be served by thrombin activation in brain? Could we derive insights about the response of the brain to injury, and specifically explore the cell-subtype interactions during injury among astrocytes, neurons, pericytes, oligodendroglia, and endothelial cells? To answer these questions, and to address the central hypothesis of the present award application, we added significant new models to our repertoire, and generated pilot data speaking to the feasibility and novelty of the proposed new specific aims.