King's College London

TY - JOUR

T1 - Proteomics of REPLICANT perfusate detects changes in the metastatic lymph node microenvironment

AU - Stevenson, Julia

AU - Barrow-McGee, Rachel

AU - Yu, Lu

AU - Paul, Angela

AU - Mansfield, David

AU - Owen, Julie

AU - Woodman, Natalie

AU - Natrajan, Rachael

AU - Haider, Syed

AU - Gillett, Cheryl

AU - Tutt, Andrew

AU - Pinder, Sarah E.

AU - Choudary, Jyoti

AU - Naidoo, Kalnisha

N1 - Funding Information: The authors would like to thank the patients for consenting for use of their tissue to develop this model, and the surgeons who assisted with this. This work was funded by Cancer Research UK (Pioneer Award CRC598X). We thank Breast Cancer Now for funding this work as part of Programme Funding to the Breast Cancer Now Toby Robins Research Centre. This work was funded by a Clinical Lectureship at The Institute of Cancer Research from the National Institute of Health Research (NIHR) to Dr Kalnisha Naidoo. This study represents independent research supported by the NIHR Biomedical Research Centre (BRC) at the Royal Marsden NHS Foundation Trust and The Institute of Cancer Research, London. The views expressed are those of the authors and not necessarily those of the NIHR or the Department of Health and Social Care. Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/12

Y1 - 2021/12

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85102187967&partnerID=8YFLogxK

U2 - 10.1038/s41523-021-00227-7

DO - 10.1038/s41523-021-00227-7

M3 - Article

AN - SCOPUS:85102187967

VL - 7

JO - npj Breast Cancer

JF - npj Breast Cancer

SN - 2374-4677

IS - 1

M1 - 24

ER -