胶质母细胞瘤（GBM）是一种致命的原发性成人恶性肿瘤，由于血脑屏障（BBB）、血肿瘤屏障（BTB）和免疫抑制肿瘤微环境（TME）的限制性，很难治疗。由于脉冲聚焦超声 (pFUS) 目前用于改善跨越这些屏障的治疗递送，本研究旨在表征 pFUS 在打开 BBB 和 BTB 后对 TME 蛋白质组学的影响。我们利用 MRI 引导的 pFUS 与超声对比微泡 (此处称为“pFUS”）选择性地短暂打开 BBB 和 BTB，研究 TME 中的蛋白质组修饰。利用原位同种异体移植小鼠 GL26 GBM 模型 (Ccr2RFP/wt - Cx3cr1GFP/wt)，使用 Luminex 48 重检测评估 pFUS 对神经胶质瘤蛋白质组学的影响。pFUS 治疗的肿瘤表现出促炎细胞因子、趋化因子和营养因子的增加（CCTF）。肿瘤中的蛋白质组变化往往在单次 pFUS 疗程 (1x) 后 24 小时达到峰值，然后水平在随后的 24 小时内趋于稳定或下降。接受三次 pFUS 治疗 (3x) 的肿瘤显示 CCTF 水平升高，早在第三次治疗后 6 小时就达到峰值。pFUS 与微泡一起在小鼠 GBM 肿瘤的 TME 中诱导无菌炎症反应。此外，这种促炎性转变可以持续，并且可能为多次 pFUS 后更快的反应做好准备。这些发现提出了一个有趣的潜力，即 pFUS 诱导的 BBB 和 BTB 打开不仅可以有效促进治疗药物的递送，而且还可以用来修改 TME，以协助免疫疗法克服 GBM 中的免疫逃逸。© 2024。作者（ s)，获得 Springer Science Business Media, LLC（Springer Nature 的一部分）的独家许可。

Glioblastoma (GBM), a lethal primary adult malignancy, is difficult to treat because of the restrictive nature of the blood-brain barrier (BBB), blood-tumor barrier (BTB), and the immunosuppressive tumor microenvironment (TME). Since pulsed focused ultrasound (pFUS) is currently used to improve therapeutic deliveries across these barriers, this study aims to characterize the impact of pFUS on the TME proteomics upon opening the BBB and BTB.We utilized MRI-guided, pFUS with ultrasound contrast microbubbles (termed 'pFUS' herein) to selectively and transiently open the BBB and BTB investigating proteomic modifications in the TME. Utilizing an orthotopically-allografted mouse GL26 GBM model (Ccr2RFP/wt - Cx3cr1GFP/wt), pFUS's effect on glioma proteomics was evaluated using a Luminex 48-plex assay.pFUS treated tumors exhibited increases in pro-inflammatory cytokines, chemokines, and trophic factors (CCTFs). Proteomic changes in tumors tend to peak at 24 h after single pFUS session (1x), with levels then plateauing or declining over the subsequent 24 h. Tumors receiving three pFUS sessions (3x) showed elevated CCTFs levels peaking as early as 6 h after the third session.pFUS together with microbubbles induces a sterile inflammatory response in the TME of a mouse GBM tumor. Moreover, this proinflammatory shift can be sustained and perhaps primed for more rapid responses upon multiple sessions of pFUS. These findings raise the intriguing potential that pFUS-induced BBB and BTB opening may not only be effective in facilitating the therapeutic agent delivery, but also be harnessed to modify the TME to assist immunotherapies in overcoming immune evasion in GBM.© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.