A Single Cell and Proteomic Precision Medicine Approach to Glyburide Responsive Contusion Expansion in Severe Traumatic Brain Injury

几十年来，在翻译关于挫伤扩大机制的临床前工作方面一直存在严重的空白。 从创伤性脑损伤(TBI)到改善预后的临床治疗。这一点很重要，因为挫伤 扩张是导致创伤性脑损伤不良结局的主要原因，发病率和死亡率增加了5倍。 目前还没有治疗方法或生物标记物来识别有风险的患者。有巨大的潜力来解决这个问题 问题是，与原发损伤不同，挫伤扩张是由宿主对初始脑损伤的反应引起的，因此 是一种可改变的继发性损伤。基于指南的护理在很大程度上使用了反应性模板方法 复杂的过程，没有解决贡献途径中的个体差异；它不会阻止或限制 挫伤扩大并努力减轻危及生命的后果。这种同质化战略 不出所料，这种异质疾病导致了许多失败的临床试验。我们的长期目标是 利用相关的个体数据(分子、单细胞[SC]、基因、成像)来指导精确医学 颅脑挫伤扩大。R21解决了挫伤治疗中存在的现有知识空白 正在为转换做准备的扩展：格列本脲(Gly)。现有的研究产生了令人兴奋的势头，但 还揭示了GLY靶标的主要个体差异，这些差异可能会影响药物反应/成功翻译。 我们的目标是使用SC和蛋白质组学策略来分子内型Gly靶向途径 人类颅脑损伤中的挫伤扩展。其基本原理是，它使我们能够更好地理解不同的好处 GLY和优化翻译的机会：它告知关键的目标和可测量的细胞起源 挫伤扩张路径。中心假设是可量化的细胞类型特异性的子集 GLY靶向的差异表达的基因、途径和蛋白确定了TBI挫伤扩张的风险。 目的1证明脑脊液(CSF)SC转录信号内型Gly靶向 人类的挫伤扩大。目标2表明挫伤扩张是在GLY靶向之前的 蛋白质生物标记物发生变化。目标是协同的：细胞型差异基因表达(AIM 1)告知 挫伤扩大的可测量脑脊液生物标志物的可能来源(目标2)。考虑到令人振奋，这项工作是可行的 试点数据，现有的TBI生物库，一个已建立的多学科团队和生物信息管道。它是 创新，因为它转变了一种基于指南的精准医学方法，创建了首个人类脑脊液地图集 在脑损伤中的SC反应，并识别被测试药物靶向的通路中的挫伤扩张生物标记物 在人类脑损伤中。预期的影响包括基于分子内型的风险分层和丰富的患者- GLY试验的选择(高风险，药效学反应)。导致挫伤的独特细胞成分 扩增结合早期临床可测量的脑脊液生物标记物可以引导前所未有的细胞和靶点 精确治疗，包括新的(预防性的)可用药靶点。这为范式转换奠定了基础 基于SC的精确医学方法，以了解、监测和治疗颅脑损伤中毁灭性的继发性损伤。

For decades, there has been a critical gap in translating preclinical work on mechanisms of contusion expansion in traumatic brain injury (TBI) to clinical therapies that improve outcome. This is important because contusion expansion is a major driver of unfavorable outcome in TBI with up to 5X increase in morbidity and mortality, yet there are no treatments or biomarkers to identify patients at risk. There is immense potential to address this issue because unlike primary injury, contusion expansion results from host response to the initial TBI and thus is a modifiable secondary injury. Guideline-based care uses a reactive templated approach to this hugely complex process without addressing individual differences in contributory pathways; it does not prevent or limit contusion expansion and struggles to mitigate the life-threatening consequences. Such homogeneous strategies for a heterogeneous disease have unsurprisingly led to many failed clinical trials. Our long-term goal is to harness relevant individual data (molecular, single-cell [SC], genetic, imaging) to direct precision medicine for TBI contusion expansion. This R21 addresses existing knowledge gaps in a promising therapy for contusion expansion being primed for translation: Glyburide (GLY). Existing research generated exciting momentum but also revealed major individual differences in GLY targets that could affect drug-response/successful translation. Our objective is to use SC and proteomic strategies to molecularly endotype GLY-targeted pathways of contusion expansion in human TBI. The rationale is that it allows us to better understand heterogeneous benefits and opportunities of GLY and optimize translation: it informs cellular origins of key targetable and measurable contusion expansion pathways. The central hypothesis is that a subset of quantifiable cell-type specific differentially expressed genes, pathways and proteins targeted by GLY identify risk for TBI contusion expansion. Aim 1 demonstrates that cerebrospinal fluid (CSF) SC transcriptomic signatures endotype GLY-targeted contusion expansion in humans. Aim 2 demonstrates that contusion expansion is preceded by GLY-targetable protein biomarkers changes. The aims are synergistic: cell-type differential gene expression (Aim 1) informs likely sources of measurable CSF biomarkers (Aim 2) of contusion expansion. The work is feasible given exciting pilot data, an existing TBI biobank, an established multidisciplinary team and bioinformatic pipelines. It is innovative as it shifts a guideline-based approach to precision medicine, creates a first-in-human atlas of CSF SC response in TBI, and identifies contusion expansion biomarkers in pathways targeted by a drug being tested in human TBI. The expected impact includes molecular endotype-based risk-stratification and enriched patient- selection for GLY trials (high risk, pharmacodynamic response). Unique cellular components that drive contusion expansion combined with early clinically measurable CSF biomarkers can guide unprecedented cell- and target- precise therapy including novel (preventive) druggable targets. This lays the foundation for a paradigm shifting SC-based precision medicine approach to understand, monitor, and treat a devastating secondary injury in TBI.