Stevenson Julia, Barrow-McGee Rachel, Yu Lu, Paul Angela, Mansfield David, Owen Julie, Woodman Natalie, Natrajan Rachael, Haider Syed, Gillett Cheryl, Tutt Andrew, Pinder Sarah E, Choudary Jyoti, Naidoo Kalnisha
The Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK.
Division of Cancer Biology, The Institute of Cancer Research, London, UK.
NPJ Breast Cancer. 2021 Mar 5;7(1):24. doi: 10.1038/s41523-021-00227-7.
In breast cancer (BC), detecting low volumes of axillary lymph node (ALN) metastasis pre-operatively is difficult and novel biomarkers are needed. We recently showed that patient-derived ALNs can be sustained ex-vivo using normothermic perfusion. We now compare reactive (tumour-free; n = 5) and macrometastatic (containing tumour deposits >2 mm; n = 4) ALNs by combining whole section multiplex immunofluorescence with TMT-labelled LC-MS/MS of the circulating perfusate. Macrometastases contained significantly fewer B cells and T cells (CD4/CD8/regulatory) than reactive nodes (p = 0.02). Similarly, pathway analysis of the perfusate proteome (119/1453 proteins significantly differentially expressed) showed that immune function was diminished in macrometastases in favour of 'extracellular matrix degradation'; only 'neutrophil degranulation' was preserved. Qualitative comparison of the perfusate proteome to that of node-positive pancreatic and prostatic adenocarcinoma also highlighted 'neutrophil degranulation' as a contributing factor to nodal metastasis. Thus, metastasis-induced changes in the REPLICANT perfusate proteome are detectable, and could facilitate biomarker discovery.
在乳腺癌（BC）中，术前检测低体积腋窝淋巴结（ALN）转移很困难，因此需要新的生物标志物。我们最近发现，使用常温灌注可以在体外维持患者来源的ALN。现在，我们通过将全切片多重免疫荧光与循环灌注液的TMT标记液相色谱-质谱联用，比较反应性（无肿瘤；n = 5）和大转移（含有>2 mm肿瘤沉积物；n = 4）的ALN。与反应性淋巴结相比，大转移灶中的B细胞和T细胞（CD4/CD8/调节性）明显更少（p = 0.02）。同样，灌注液蛋白质组的通路分析（119/1453种蛋白质有显著差异表达）表明，大转移灶中的免疫功能减弱，有利于“细胞外基质降解”；仅“中性粒细胞脱颗粒”得以保留。将灌注液蛋白质组与淋巴结阳性的胰腺癌和前列腺癌的蛋白质组进行定性比较，也突出了“中性粒细胞脱颗粒”是淋巴结转移的一个促成因素。因此，转移诱导的REPLICANT灌注液蛋白质组变化是可检测的，并且有助于生物标志物的发现。
