宫颈癌是女性生殖系统恶性肿瘤中死亡率第二高的癌症。程序性细胞死亡蛋白1（PD-1）阻断剂等免疫检查点抑制剂是有前途的治疗药物，但其与新辅助化疗（NACT）联合使用时的疗效尚未得到充分测试，并且它们如何改变肿瘤微环境尚未得到全面阐明在这项研究中，我们使用来自九种人类宫颈癌组织的 46,950 个细胞进行了单细胞 RNA 测序，这些细胞代表了 NACT 和 PD-1 阻断联合疗法的连续不同阶段。我们描绘了宫颈上皮细胞的轨迹，并确定了联合治疗中涉及的关键因素。在肿瘤细胞和免疫细胞之间进行细胞间通讯分析。此外，将THP-1来源和原代单核细胞来源的巨噬细胞与宫颈癌细胞共培养，并通过流式细胞术检测吞噬作用。使用CD74人源化皮下肿瘤模型在体内验证了阻断CD74的抗肿瘤活性。通路富集分析表明NACT激活了细胞因子和补体相关的免疫反应。细胞间通讯分析显示，NACT治疗后，T细胞和癌细胞之间的相互作用强度下降，但巨噬细胞和癌细胞之间的相互作用强度增强。我们证实巨噬细胞是 PD-1 阻断在体外发挥抗肿瘤作用所必需的。此外，在联合 NACT 期间，CD74 阳性巨噬细胞经常与免疫反应性最强的上皮亚组 3 (Epi3) 癌症亚组相互作用。我们发现CD74的上调限制了吞噬作用并刺激了M2极化，而CD74的阻断则增强了巨噬细胞的吞噬作用，降低了宫颈癌细胞在体外和体内的活力。我们的研究揭示了结合NACT和NACT对宫颈癌微环境中动态细胞-细胞相互作用网络的影响PD-1阻断。此外，阻断肿瘤相关巨噬细胞衍生的 CD74 可以增强新辅助治疗效果。© 作者（或其雇主）2024。CC BY-NC 允许重复使用。禁止商业再利用。请参阅权利和权限。英国医学杂志出版。

Cervical cancer has the second-highest mortality rate among malignant tumors of the female reproductive system. Immune checkpoint inhibitors such as programmed cell death protein 1 (PD-1) blockade are promising therapeutic agents, but their efficacy when combined with neoadjuvant chemotherapy (NACT) has not been fully tested, and how they alter the tumor microenvironment has not been comprehensively elucidated.In this study, we conducted single-cell RNA sequencing using 46,950 cells from nine human cervical cancer tissues representing sequential different stages of NACT and PD-1 blockade combination therapy. We delineated the trajectory of cervical epithelial cells and identified the crucial factors involved in combination therapy. Cell-cell communication analysis was performed between tumor and immune cells. In addition, THP-1-derived and primary monocyte-derived macrophages were cocultured with cervical cancer cells and phagocytosis was detected by flow cytometry. The antitumor activity of blocking CD74 was validated in vivo using a CD74 humanized subcutaneous tumor model.Pathway enrichment analysis indicated that NACT activated cytokine and complement-related immune responses. Cell-cell communication analysis revealed that after NACT therapy, interaction strength between T cells and cancer cells decreased, but intensified between macrophages and cancer cells. We verified that macrophages were necessary for the PD-1 blockade to exert antitumor effects in vitro. Additionally, CD74-positive macrophages frequently interacted with the most immunoreactive epithelial subgroup 3 (Epi3) cancer subgroup during combination NACT. We found that CD74 upregulation limited phagocytosis and stimulated M2 polarization, whereas CD74 blockade enhanced macrophage phagocytosis, decreasing cervical cancer cell viability in vitro and in vivo.Our study reveals the dynamic cell-cell interaction network in the cervical cancer microenvironment influenced by combining NACT and PD-1 blockade. Furthermore, blocking tumor-associated macrophage-derived CD74 could augment neoadjuvant therapeutic efficacy.© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.