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Proteomics of REPLICANT Perfusate Detects Changes in the Metastatic Lymph Node Microenvironment

ABSTRACT: n 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 >2mm; n = 4) ALNs by combining whole section multiplex immunofluorescence with TMT-labelled LC-MS/MS of the circulating perfusate. Macrometastases contained significantly fewer B- 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.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Lymph Node

DISEASE(S): Breast Cancer

SUBMITTER: James Wright

LAB HEAD: Jyoti Choudhary

PROVIDER: PXD022722 | Pride | 2021-01-05

REPOSITORIES: Pride

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	20190601_AP_FASP_NISH_Naidoo_Lab_Set1_1.raw	Raw
	20190601_AP_FASP_NISH_Naidoo_Lab_Set1_2.raw	Raw
	20190601_AP_FASP_NISH_Naidoo_Lab_Set1_3.raw	Raw
	20190601_AP_FASP_NISH_Naidoo_Lab_Set1_4.raw	Raw
	20190601_AP_FASP_NISH_Naidoo_Lab_Set1_5.raw	Raw

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