Hu Mufeng, Weldy Anna, Lovalvo Isabella, Akins Erin, Jain Saket, Chang Alexander, Sati Ankita, Lad Meeki, Lui Austin, Rajidi Akhil, Kothekar Ameya, Ding Erika, Kumar Sanjay, Aghi Manish K
University of California San Francisco (UCSF) Neurosurgery.
UC Berkeley Chemical and Biomolecular Engineering.
bioRxiv. 2025 Jan 24:2025.01.20.633978. doi: 10.1101/2025.01.20.633978.
To identify new therapeutic targets that limit glioblastoma (GBM) invasion, we applied druggable-genome CRISPR screens to patient-derived GBM cells in micro-dissectible biomimetic 3D hydrogel platforms that permit separation and independent analysis of core vs. invasive fractions. We identified 12 targets whose suppression limited invasion, of which ACP1 (LMW-PTP) and Aurora Kinase B (AURKB) were validated in neurosphere assays. Proximity labeling analysis identified cortactin as an ACP1-AURKB link, as cortactin undergoes serine phosphorylation by AURKB and tyrosine dephosphorylation by ACP1. Suppression of ACP1 or AURKB in culture and shifted the balance of cortactin phosphorylation in GBM and reduced actin polymerization and actin-cortactin co-localization. Additional biophysical analysis implicated AURKB in GBM cell adhesion and cortical stiffness, and ACP1 in resistance to mechanical stress and shape plasticity needed for 3D migration. These findings reveal a novel targetable axis that balances kinase and phosphatase activities to regulate actin polymerization during GBM invasion.
为了确定限制胶质母细胞瘤（GBM）侵袭的新治疗靶点，我们在可微解剖的仿生3D水凝胶平台上对患者来源的GBM细胞应用了可药物基因组CRISPR筛选，该平台允许分离并独立分析核心部分与侵袭部分。我们确定了12个抑制后能限制侵袭的靶点，其中酸性磷酸酶1（LMW-PTP）和极光激酶B（AURKB）在神经球试验中得到验证。邻近标记分析确定皮层肌动蛋白是ACP1-AURKB的连接点，因为皮层肌动蛋白会受到AURKB的丝氨酸磷酸化以及ACP1的酪氨酸去磷酸化作用。在培养中抑制ACP1或AURKB会改变GBM中皮层肌动蛋白磷酸化的平衡，并减少肌动蛋白聚合以及肌动蛋白与皮层肌动蛋白的共定位。进一步的生物物理分析表明，AURKB与GBM细胞黏附和皮层硬度有关，而ACP1与3D迁移所需的机械应力抗性和形状可塑性有关。这些发现揭示了一个新的可靶向轴，该轴平衡激酶和磷酸酶活性以在GBM侵袭过程中调节肌动蛋白聚合。
