Rani Anupama, Raikwar Sudhanshu P, Yoo Wonsuk, Ahmad Saif, Vagni Vincent A, Janesko-Feldman Keri L, Carlson Shaun W, Eberle Adam, Miller Margaux, Helm John, Catapano Joshua, Zusman Benjamin E, Desai Shashvat, Javelosa Raemier Anne M, Afework Semeon, McNally Erin Audrey, Kohanbash Gary, Rajasundaram Dhivyaa, Waters Michael F, Ducruet Andrew, Jadhav Ashutosh, Kumar Aditya, Phuah Chia-Ling, Kochanek Patrick M, Jha Ruchira M
Department of Translational Neuroscience, Barrow Neurological Institute and St. Joseph's Hospital and Medical Center, Phoenix, Arizona, USA.
Ivy Brain Tumor Center and Department of Translational Neuroscience, Barrow Neurological Institute, St. Joseph's Hospital and Medical Center, Phoenix, Arizona, USA.
J Neurotrauma. 2025 Aug;42(15-16):1319-1344. doi: 10.1089/neu.2024.0232. Epub 2025 Jun 12.
Sex disparities in traumatic brain injury (TBI) remain poorly understood. Previous data suggest that males are more susceptible to acute secondary injury processes and cell death, whereas females are more vulnerable chronically. Additional sex-based differences have been reported depending on injury model/severity and post-traumatic neurodegeneration. This gap in understanding limits therapy translation. We previously demonstrated sex-based differences in genetic modulation of a key pathway of secondary injury in TBI, sulfonylurea-receptor 1 (SUR1). Glibenclamide (GLI, SUR1-inhibitor) has shown promise in pre-clinical and early clinical studies of TBI and stroke. Here, we evaluated GLI's modulation of multifaceted TBI outcomes across sex for the first time. In total, 120 mice were randomized to controlled cortical impact (CCI) ± GLI or vehicle (dimethyl sulfoxide, DMSO). Either vehicle or GLI treatment was administered post-CCI using an intraperitoneal (IP) loading dose (10 µg/mouse, 10 min post-TBI), followed by a 7-day subcutaneous maintenance infusion at 0.5 µL/h using ALZET mini-osmotic pumps (1007D, Durect Corp.). Mice were tested for cognitive function (Morris water maze, MWM), motor function (rotarod), anxiety (elevated plus maze, EPM), immunofluorescence markers of neurodegeneration (TAU, TDP43), neurogenesis (SOX2, Ki67), angiogenesis (VEGFA), and cerebral blood flow (CBF) to interrogate behavioral, molecular, and physiological effects of TBI and therapy. Different measures within behavioral, immunofluorescence, and CBF outcomes varied across sex, either post-CCI and/or in response to GLI. Motor impairment had baseline differences across sex post-CCI. In both sexes, behavioral deficits were improved by GLI. The effect of GLI on behavior was moderated by sex, with greater benefit in males versus females, including improved MWM latency ( < 0.0001) and rotarod latency ( = 0.016, revolutions per minute, = 0.03). Males had increased anxiety post-CCI (EPM); GLI was beneficial across sexes. TDP43 and TAU in several brain regions were increased 72 h post-CCI (males>females, all < 0.0001). These remained markedly elevated only in females by 21 days, whereas TAU in males decreased without treatment. GLI downregulated TDP43 and TAU across sex and brain region (all < 0.01-0.0001). In females only, DMSO had similar effects as GLI on TDP43 and TAU. SOX2 was increased in the dentate gyrus (DG) only in males post-CCI ( < 0.01, < 0.001). GLI increased DG SOX2 in females ( < 0.05, < 0.001). GLI increased VEGFA at 72 h across sexes. CCI reduced CBF acutely in both sexes; in males, GLI improved this by 21 days ( = 0.031). In females, both GLI and DMSO-vehicle benefited CBF versus untreated-CCI. We demonstrate novel sex-based differences post-CCI and GLI-response across several metrics. TAU was chronically elevated (and responsive to treatment) in females, not males, potentially providing a sex-specific target. DMSO may have previously unrecognized benefits on certain pathways (TAU, CBF) in females. Although GLI has multifaceted benefits across sexes, effects are more pronounced in males. This may have important implications for clinical trial study design and analysis.
创伤性脑损伤（TBI）中的性别差异仍未得到充分理解。先前的数据表明，男性更容易受到急性继发性损伤过程和细胞死亡的影响，而女性在慢性阶段则更易受损。根据损伤模型/严重程度和创伤后神经退行性变，还报告了其他基于性别的差异。这种认知上的差距限制了治疗方法的转化。我们先前证明了在TBI继发性损伤的关键途径磺酰脲受体1（SUR1）的基因调控中存在基于性别的差异。格列本脲（GLI，SUR1抑制剂）在TBI和中风的临床前和早期临床研究中已显示出前景。在此研究中，我们首次评估了GLI对不同性别TBI多方面结果的调节作用。总共120只小鼠被随机分为控制性皮质撞击（CCI）±GLI或溶剂（二甲基亚砜，DMSO）组。在CCI后通过腹腔内（IP）负荷剂量（10μg/小鼠，TBI后10分钟）给予溶剂或GLI治疗，随后使用ALZET微型渗透泵（1007D，Durect公司）以0.5μL/h的速度进行7天的皮下维持输注。对小鼠进行认知功能（莫里斯水迷宫，MWM）、运动功能（转棒试验）、焦虑（高架十字迷宫，EPM）、神经退行性变的免疫荧光标记物（TAU、TDP43）、神经发生（SOX2、Ki67）、血管生成（VEGFA）和脑血流量（CBF）的测试，以探究TBI和治疗的行为、分子和生理效应。行为、免疫荧光和CBF结果中的不同测量指标在性别之间存在差异，无论是在CCI后和/或对GLI的反应中。CCI后运动障碍在性别之间存在基线差异。在两性中，GLI均改善了行为缺陷。GLI对行为的影响因性别而异，对男性的益处大于女性，包括改善MWM潜伏期（<0.0001）和转棒试验潜伏期（=0.016，每分钟转数，=0.03）。CCI后男性的焦虑增加（EPM）；GLI对两性均有益。CCI后72小时，几个脑区的TDP43和TAU增加（男性>女性，均<0.0001）。到21天时，这些指标仅在女性中仍显著升高，而男性的TAU未经治疗就下降了。GLI在不同性别和脑区下调了TDP43和TAU（均<0.01 - 0.0001）。仅在女性中，DMSO对TDP43和TAU的影响与GLI相似。CCI后仅在男性的齿状回（DG）中SOX2增加（<0.01，<0.001）。GLI增加了女性DG中的SOX2（<0.05，<0.001）。GLI在72小时时增加了两性的VEGFA。CCI在两性中均急性降低了CBF；在男性中，GLI在21天时改善了这一情况（=0.031）。在女性中，GLI和DMSO溶剂组相对于未治疗的CCI组均对CBF有益。我们证明了CCI后和GLI反应在几个指标上存在新的基于性别的差异。TAU在女性中慢性升高（且对治疗有反应），而在男性中没有，这可能提供了一个性别特异性靶点。DMSO可能对女性的某些途径（TAU、CBF）具有先前未被认识到的益处。尽管GLI对两性有多种益处，但对男性的影响更为明显。这可能对临床试验研究设计和分析具有重要意义。
