Dynamic Neuroimmune Profiling in Patients with Acute Intracerebral Hemorrhage

项目摘要 脑出血(ICH)是一种致死率和致残率都很高的破坏性卒中。临床前 模型已经确定了免疫反应在急性脑损伤和功能恢复中的关键作用。 然而，在人类患者身上进行的研究更具挑战性。将啮齿动物的病理生理学联系起来 在患者身上看到的模型是成功开发免疫调节疗法的关键 脑出血的治疗。 Mistie III是一项由NINDS资助的3期随机临床试验，测试微创治疗的疗效 血肿清除对脑出血后6个月和12个月功能预后的影响。试验设计提供非物质文化遗产 脑出血发病后2-5天的引流样本，正好是局部炎症反应的时间点 有助于损伤和调节反应，帮助修复啮齿动物模型。这提供了无与伦比的 回答有关存活患者脑出血病理生理学的基本生物学问题的机会结束 时间到了。 总的假设是，随着时间的推移，血液来源的巨噬细胞会转变为另一种激活方式，而这 转化是通过减少细胞因子信号，增强吞噬功能，以及 抑制T效应反应。最终，我们预测这一过渡的时机将直接影响患者 结果。 目的1将随着时间的推移确定患者大脑和外周血中的细胞免疫反应。 CD16+单核细胞、CD4+CD127+CD25-效应T细胞、CD4+CD127-CD25+调节性T细胞和CD8+ 细胞毒性T细胞将被分选，以便使用RNA-seq进行深入的转录分析。补充性研究将 利用质量细胞术全面定义和表型白细胞群并跟踪细胞 随着时间的推移种群数量。免疫反应的变化将利用系统生物学进行整合 将确定与患者预后相关的方法和关联。 目标2将确定变化的白细胞表型如何决定巨噬细胞的效应功能和 T细胞。使用从患者血液和脑组织中获取的巨噬细胞和T细胞进行体外检测 不同的时间点，吞噬和抑制效应T细胞反应的分子机制将 被指认出来。

Project Summary Intracerebral hemorrhage (ICH) is a devastating type of stroke with high mortality and morbidity. Preclinical models have identified key roles for immune responses in both acute brain injury and in functional recovery. However, studies in human patients have been more challenging. Linking the pathophysiology seen in rodent models to that seen in patients is critical to the successful development of immunomodulatory therapies for the treatment of ICH. MISTIE III is a NINDS-funded, phase-3 randomized clinical trial testing the efficacy of minimally invasive hematoma evacuation on functional outcomes at 6 and 12 months after ICH. The trial design provides ICH drainage samples over days 2-5 after ICH onset, precisely the time points that local inflammatory responses contribute to injury and modulate responses that aid in repair in rodent models. This provides an unparalleled opportunity to answer fundamental biological questions about the pathophysiology of ICH in living patients over time. The overall hypothesis is that blood-derived macrophages transition to alternative activation over time and this transition is critical to resolution of inflammation via reduced cytokine signaling, enhanced phagocytosis, and inhibition of T effector responses. Ultimately, we predict that the timing of this transition directly impacts patient outcome. Aim 1 will determine the cellular immune responses in the brain and peripheral blood of patients over time. CD16+ monocytes, CD4+CD127+CD25- effector T cells, CD4+CD127-CD25+ regulatory T cells, and CD8+ cytotoxic T cells will be sorted for in depth transcriptional profiling using RNA-seq. Complementary studies will utilize mass cytometry to comprehensively define and phenotype the leukocyte populations and track cell populations over time. Changes in the immune responses will be integrated using systems biology approaches and associations with patient outcome will be identified. Aim 2 will determine how changing leukocyte phenotypes determine the effector functions of macrophages and T cells. Using ex vivo assays using macrophages and T cells harvested from patients blood and brain at various time points, the molecular mechanisms of phagocytosis and inhibition of effector T cell responses will be identified.