Potential stroke therapeutic efficacy of FumET-CORM through the Nrf2 pathway

我们和其他人一直是靶向转录因子Nrf 2的先驱，Nrf 2是氧化应激的主要调节因子。 应激和炎症，作为中风治疗的一个有前途的策略。我们和其他人也记录了 CO和“CO供体”可以提供神经保护的可能性。私家侦探实验室的证据显示 小分子，如富马酸二甲酯（DMF），一种典型的Nrf 2诱导剂正在临床上使用， 低剂量的一氧化碳，与传统的有毒物质观点相反，显示出持续的神经保护作用。 抗缺血性脑损伤和功能缺陷的功效，特别是通过Nrf 2途径。 因此，我们可以预测，含有富马酸甲酯和CO释放的分子 分子单位（即FumET-CORM）可能具有有趣和有前途的治疗潜力。我们 初步结果表明，CO或DMF暴露对短暂性和永久性病灶均有保护作用 脑缺血小鼠模型。FumET-CORM已被证明具有抗炎作用， 财产这促使我们假设新的FumET-CORM将保护免受 永久性远端大脑中动脉闭塞（pdMCAO）后的神经功能缺损和梗死;以及 这种有益作用至少部分地由Nrf 2细胞保护途径调节。 在AIM 1中，我们将确定FumET-CORM（与其无活性的iFumET-CORM相比）是否对以下疾病具有保护作用： 神经缺陷和脑损伤。我们将测试剂量反应和治疗 窗口方案，以确定雄性和雌性成年小鼠的最佳有益剂量方案。初级 结局将基于神经/认知变化，并将进行各种IHC染色， 探索各种假定途径的含义。还将监测毒性。最优 将在老年同窝仔中扩展剂量/治疗方案。 在AIM 2中，我们将研究Nrf 2通路是否在FumET-CORM“双重”中起重要作用。 行动药物”的有益机制，并开始解决哪些脑细胞类型（S）负责这种 通过首先使用共免疫染色方法来进行神经保护。使用目标1中的最佳条件，我们将 使用整体Nrf 2-/-C57 BL/6小鼠评估Nrf 2通路的贡献，并讨论哪种脑细胞 类型揭示了最深刻的Nrf 2相关变化。这将决定未来的测试， 合适的诱导型Nrf 2基因工程cre小鼠-使用最新和最具选择性的cre小鼠。 总之，我们将测试的假设，一些估算的有益效果的FumET-CORM “双重作用药物”对脑损伤的作用至少部分归因于Nrf 2，而哪些细胞可以最 负责假定的神经保护。这可以用于缺血性中风患者， 潜在的其他急性脑损伤。我们现在需要这种R21机制来获得现在所需的数据 为了测试这个工作假设，我们有团队和专业知识来完成这个2年的提案。

We and others have been pioneers in targeting the transcriptional factor Nrf2, a master regulator of oxidative stress and inflammation, as a promising strategy for stroke therapy. We and others have also documented the possibility that CO and “CO donor” can provide neuroprotection. Accumulated evidence from the PI's lab show that small molecules such as the dimethyl fumarate (DMF), a typical Nrf2 inducer being used in the clinic, and CO at low doses, contrary to the traditional view as a toxic agent, have displayed sustained neuroprotective efficacy against ischemic brain damage and functional deficits notably via the Nrf2 pathway. Consequently, we would predict that a molecule containing both a methyl fumarate and CO-releasing molecule unit (i.e. FumET-CORM) could have an intriguing and promising therapeutic potential. Our preliminary results showed that CO or DMF exposure is protective in both transient and permanent focal cerebral ischemia mouse models. FumET-CORM has already been shown to exhibit anti-inflammatory property. This prompts us to work on the hypothesis that the new FumET-CORM would be protective against neurological deficits and infarct after permanent distal middle cerebral artery occlusion (pdMCAO); and that this beneficial effect would be, at least partially, regulated by the Nrf2 cytoprotective pathway. In AIM 1, we will determine whether FumET-CORM (vs its inactive iFumET-CORM) is protective against neurological deficits and brain damage following pdMCAO. We will test a dose response and a therapeutic window protocols to determine the optimal beneficial dose regimen in male and female adult mice. Primary outcomes will be based on neurological/cognitive changes, and various IHC staining will be performed to start exploring the implications of various putative pathways. Toxicity will also be monitored. The optimal dose/treatment regimen will be extended in the aged littermates. In AIM 2, we will examine whether the Nrf2 pathway plays a significant role in the FumET-CORM “dual- action drug” beneficial mechanism and start addressing which brain cell type(s) is responsible for such neuroprotection by first using a co-immunostaining approach. Using the optimal conditions from Aim 1, we will evaluate the contribution of Nrf2 pathway using global Nrf2-/- C57BL/6 mice, and also address which brain cell type(s) reveal the most profound Nrf2-related changes. This would then dictate the future test of the most appropriate inducible Nrf2 flox-cre mice – using the latest and most selective cre mice. Together, we will test the hypothesis that some of the imputed beneficial effects of the FumET-CORM “dual-action drug” on brain damage are attributed at least in part to Nrf2 and which cells can be most responsible for the putative neuroprotection. This could be administered to ischemic stroke patients and potentially other acute brain insults. This R21 mechanism is now necessary us to obtain the data now required to test this working hypothesis, and we have the team and expertise to accomplish this 2yr-proposal.