Understanding how breast cancer (BC) grows in axillary lymph nodes (ALNs), and refining how therapies might halt that process, is clinically important. However, modelling the complex ALN microenvironment is difficult, and no human models exist at present. We harvested ALNs from ten BC patients, and perfused them at 37 °C ex vivo for up to 24 h. Controlled autologous testing showed that ALNs remain viable after 24 h of ex vivo perfusion: haematoxylin and eosin-stained histological appearance and proliferation (by Ki67 immunohistochemistry) did not change significantly over time for any perfused ALN compared with a control from time-point zero. Furthermore, targeted gene expression analysis (NanoString PanCancer IO360 panel) showed that only 21/750 genes were differentially expressed between control and perfused ALNs (|log2 FC| > 1 and q < 0.1): none were involved in apoptosis and metabolism, but rather all 21 genes were involved in immune function and angiogenesis. During perfusion, tissue acid-base balance remained stable. Interestingly, the flow rate increased (p < 0.001) in cancer-replaced (i.e. metastasis occupied more than 90% of the surface area on multiple levels) compared to cancer-free nodes (i.e. nodes with no metastasis on multiple sections). CXCL11 transcripts were significantly more abundant in cancer-replaced nodes, while CXCL12 transcripts were significantly more abundant in cancer-free nodes. These cytokines were also detected in the circulating perfusate. Monoclonal antibodies (nivolumab and trastuzumab) were administered into a further three ALNs to confirm perfusion efficacy. These drugs saturated the nodes; nivolumab even induced cancer cell death. Normothermic ALN perfusion is not only feasible but sustains the tumour microenvironment ex vivo for scientific investigation. This model could facilitate the identification of actionable immuno-oncology targets. © 2019 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

中文翻译：

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实时侵入人体淋巴瘤的实时浸润灌注（癌症）：一项可行性研究。

了解乳腺癌（BC）如何在腋窝淋巴结（ALN）中生长，以及完善疗法如何阻止该过程在临床上很重要。但是，很难对复杂的ALN微环境进行建模，并且目前还没有人类模型。我们从十名卑诗省患者中收集了ALN，并在37°C下离体灌注了长达24小时。对照自体测试显示离体灌注24小时后，ALN仍保持活力：苏木精和曙红染色的组织学外观和增殖（通过Ki67免疫组织化学）随时间的变化与零时点的对照相比均无显着变化。此外，靶向基因表达分析（NanoString PanCancer IO360面板）显示，对照和灌注ALN之间仅差异表达21/750个基因（| log2 FC |> 1和q <0）。1）：没有一个参与细胞凋亡和代谢，但是所有21个基因都参与了免疫功能和血管生成。在灌注期间，组织酸碱平衡保持稳定。有趣的是，与无癌的淋巴结（即在多个部位无转移的淋巴结）相比，在被癌症替换的淋巴结（即，转移在多个层面上占表面积的90％以上）的流速增加（p <0.001）。CXCL11转录本在癌症置换淋巴结中显着丰富，而CXCL12转录本在无癌淋巴结中显着更为丰富。在循环灌流液中也检测到了这些细胞因子。将单克隆抗体（nivolumab和曲妥珠单抗）施用于另外三个ALN中，以确认灌注功效。这些药物使淋巴结饱和。尼武单抗甚至诱导癌细胞死亡。正常体温的ALN灌注不仅可行，而且可以离体维持肿瘤的微环境，以进行科学研究。该模型可以促进可操作的免疫肿瘤靶标的鉴定。©2019作者。John Wiley＆Sons Ltd代表大不列颠及爱尔兰病理学会出版的《病理学杂志》。