Enhancing Tumor Targeting of Antitumor Drugs through Anlotinib-Mediated Modulation of Extracellular Matrix and RhoA/ROCK Signaling Pathway

In this study, our research focused on investigating the impact of Anlotinib in enhancing the tumor targeting of antitumor drugs in vivo. Through RNA-sequencing and Label-free quantitative proteomics analysis, we discovered that Anlotinib effectively regulated the expression of extracellular matrix (ECM) genes and proteins, leading to a significant reduction in ECM stiffness. Our bioinformatic analysis indicated a potential positive relationship between the ECM pathways and gefitinib resistance, poor PD-1 treatment outcomes, as well as unfavorable prognosis in lung cancer patients following chemotherapy. To evaluate the efficacy of Anlotinib, we administered it in combination with anti-PD-1/PD-L1 agents, chemotherapeutic drugs, and gefitinib, and visualized their distribution using fluorescent labeling in various tumor types. Notably, our results demonstrated that Anlotinib substantially improved the drug targeting process by prolonging the retention time of antitumor drugs at the tumor site. Moreover, the combination therapy induced a notable loosening of the tumor tissue structure compared to the control group. This reduction in stiffness was further associated with decreased interstitial fluid pressures and tumor solid pressure. Additionally, we observed that Anlotinib effectively suppressed the RhoA/ROCK signaling pathway, inhibiting the formation of stress fibers. These findings strongly suggest that Anlotinib enhances the distribution and retention of antitumor drugs in tumors by modulating ECM expression and physical properties through the RhoA/ROCK signaling pathway. These valuable insights contribute to the development of combination therapies aimed at improving tumor targeting in cancer treatment. Overall design: In order to detect what pathways Anlotinib depends on affecting the hydraulic and solid pressure of tumor stroma. We established a cell line A549 treated with Anlotinib. We then performed gene expression profiling analysis using date obtained from RNA-seq of A549 cells treated with Anlotinib or DMSO for 48 h

本研究聚焦于探究安罗替尼（Anlotinib）在体内增强抗肿瘤药物肿瘤靶向性的作用效果。通过RNA测序（RNA-sequencing）与无标记定量蛋白质组学（Label-free quantitative proteomics）分析，我们发现安罗替尼可有效调控细胞外基质（ECM）基因与蛋白的表达，显著降低ECM硬度。我们的生物信息学分析显示，ECM通路与吉非替尼（gefitinib）耐药、PD-1治疗效果不佳以及肺癌患者化疗后预后不良存在潜在正相关关系。为评估安罗替尼的疗效，我们将其与抗PD-1/PD-L1制剂、化疗药物及吉非替尼联合给药，并通过荧光标记技术可视化其在多种肿瘤类型中的分布情况。值得注意的是，本研究结果证实，安罗替尼通过延长抗肿瘤药物在肿瘤部位的滞留时间，显著优化了药物靶向过程。此外，与对照组相比，联合疗法可显著松解肿瘤组织结构。这种硬度降低还与肿瘤组织间隙液压及实体压的下降相关。同时，我们观察到安罗替尼可有效抑制RhoA/ROCK信号通路，进而抑制应力纤维的形成。上述研究结果充分表明，安罗替尼可通过RhoA/ROCK信号通路调控ECM的表达与物理特性，从而增强抗肿瘤药物在肿瘤内的分布与滞留效果。这些极具价值的研究发现为开发旨在提升癌症治疗中肿瘤靶向性的联合疗法提供了重要依据。整体实验设计：为明确安罗替尼影响肿瘤间质液压与实体压所依赖的信号通路，我们构建了经安罗替尼处理的A549细胞系。随后，我们利用经安罗替尼或二甲基亚砜（DMSO）处理48小时的A549细胞的RNA测序数据，开展基因表达谱分析。