许多肿瘤中都观察到大量的肿瘤相关巨噬细胞（TAM）浸润，它们通常呈现免疫抑制的M2型表型，但也可以由于其高度的可塑性而转化为M1型的抗肿瘤表型。巨噬细胞极化状态与细胞形态的改变相关，巨噬细胞形态与激活状态相关。M1型巨噬细胞呈大而圆的形态，而M2型巨噬细胞则呈拉伸和延长的形态。研究发现通过调控细胞形态可以影响巨噬细胞的极化状态。细胞形态的形成在很大程度上依赖于细胞骨架蛋白，尤其是微管。文中提到的vinblastine（VBL）是一种微管靶向药物，被用于化疗。然而，迄今为止还没有研究探讨VBL对TAM形态变化的影响以及其在肿瘤免疫应答中的作用。我们使用荧光染色法对细胞骨架进行定量分析，揭示了M0、M1、M2、TAM和VBL处理后的TAM之间的形态学差异。流式细胞术用于使用基于细胞表面标记物的分类来确认这些巨噬细胞的极化状态。我们进行了体内抗体耗尽实验，以测试巨噬细胞和CD8+ T细胞群体对VBL的抗肿瘤效应的要求。随后，我们进行了VBL和抗PD-1联合治疗与单药治疗的比较。通过TAM的RNA-seq分析，我们探究了VBL处理前后途径活性的变化。我们使用siRNA介导的knockdown实验证实了VBL处理影响的靶点通路。我们的研究结果表明，VBL这种破坏微管的抗肿瘤药物，在体外和肿瘤模型中均能促使巨噬细胞极化为M1型表型。在缺乏巨噬细胞或CD8+ T细胞的情况下，VBL的抗肿瘤效果减弱。在机制上，VBL诱导NF-κB和Cyba依赖的活性氧自由基生成，从而使TAM极化为M1型表型。与此同时，VBL促进转录因子EB的核内转位，诱导溶酶体生物合成，并显著增加巨噬细胞的吞噬活性。本研究探讨了通过调控细胞形态影响巨噬细胞极化并从而诱导抗肿瘤反应的可能性。我们的数据揭示了VBL之前未被认识的抗肿瘤机制，并提出了将VBL与免疫检查点抑制剂相结合的药物再利用策略，以改善恶性肿瘤免疫治疗。© 本文作者（或其雇主）2023。在CC BY-NC下允许重新使用。不允许商业再使用。请参阅权限和许可。由BMJ出版。

Massive tumor-associated macrophage (TAM) infiltration is observed in many tumors, which usually display the immune-suppressive M2-like phenotype but can also be converted to an M1-like antitumor phenotype due to their high degree of plasticity. The macrophage polarization state is associated with changes in cell shape, macrophage morphology is associated with activation status. M1 macrophages appeared large and rounded, while M2 macrophages were stretched and elongated cells. Manipulating cell morphology has been shown to affect the polarization state of macrophages. The shape of the cell is largely dependent on cytoskeletal proteins, especially, microtubules. As a microtubule-targetting drug, vinblastine (VBL) has been used in chemotherapy. However, no study to date has explored the effect of VBL on TAM shape changes and its role in tumor immune response.We used fluorescent staining of the cytoskeleton and quantitative analysis to reveal the morphological differences between M0, M1, M2, TAM and VBL-treated TAM. Flow cytometry was used to confirm the polarization states of these macrophages using a cell surface marker-based classification. In vivo antibody depletion experiments in tumor mouse models were performed to test whether macrophages and CD8+ T cell populations were required for the antitumor effect of VBL. VBL and anti-PD-1 combination therapy was then investigated in comparison with monotherapy. RNA-seq of TAM of treated and untreated with VBL was performed to explore the changes in pathway activities. siRNA mediated knockdown experiments were performed to verify the target pathway that was affected by VBL treatment.Here, we showed that VBL, an antineoplastic agent that destabilizes microtubule, drove macrophage polarization into the M1-like phenotype both in vitro and in tumor models. The antitumor effect of VBL was attenuated in the absence of macrophages or CD8+ T cells. Mechanistically, VBL induces the activation of NF-κB and Cyba-dependent reactive oxygen species generation, thus polarizing TAMs to the M1 phenotype. In parallel, VBL promotes the nuclear translocation of transcription factor EB, inducing lysosome biogenesis and a dramatic increase in phagocytic activity in macrophages.This study explored whether manipulating cellular morphology affects macrophage polarization and consequently induces an antitumor response. Our data reveal a previously unrecognized antitumor mechanism of VBL and suggest a drug repurposing strategy combining VBL with immune checkpoint inhibitors to improve malignant tumor immunotherapy.© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.