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Proteomics of REPLICANT perfusate detects changes in the metastatic lymph node microenvironment
Julia Stevenson1, Rachel Barrow-McGee1, Lu Yu2
1The Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK.
NPJ Breast Cancer
|March 6, 2021
Summary
Detecting breast cancer metastasis in lymph nodes is challenging. This study found fewer immune cells and altered protein profiles in metastatic lymph nodes, suggesting new biomarker possibilities.
Area of Science:
- Oncology
- Immunology
- Proteomics
Background:
- Pre-operative detection of low-volume axillary lymph node (ALN) metastasis in breast cancer (BC) remains a clinical challenge.
- Novel biomarkers are crucial for improving diagnostic accuracy and patient outcomes.
- Previous work established ex-vivo culture of patient-derived ALNs using normothermic perfusion.
Purpose of the Study:
- To compare the cellular and proteomic profiles of reactive (tumor-free) and macrometastatic (tumor deposits >2 mm) ALNs.
- To investigate the potential of perfusate proteome analysis for biomarker discovery in ALN metastasis.
Main Methods:
- Ex-vivo normothermic perfusion of patient-derived ALNs (n=5 reactive, n=4 macrometastatic).
- Whole section multiplex immunofluorescence for cellular analysis.
- TMT-labelled liquid chromatography-tandem mass spectrometry (LC-MS/MS) for perfusate proteome analysis.
Main Results:
- Macrometastatic ALNs exhibited significantly reduced B cells and T cells (CD4+, CD8+, regulatory) compared to reactive nodes (p=0.02).
- Perfusate proteome analysis revealed diminished immune function and increased extracellular matrix degradation in macrometastatic nodes.
- 'Neutrophil degranulation' was preserved in macrometastatic nodes and identified as a common factor across different cancer types.
Conclusions:
- Metastasis induces detectable changes in the ALN perfusate proteome.
- These proteomic alterations hold potential for the discovery of novel biomarkers for axillary lymph node metastasis in breast cancer.
- The findings suggest 'neutrophil degranulation' may play a role in nodal metastasis across various cancers.
