Depleted polymorphonuclear leukocytes in human metastatic liver reflect an altered immune microenvironment associated with recurrent metastasis
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Hepatic immunity, normally protective against neoplasia, is subverted in colorectal liver metastasis (CRLM). Here, we compare the inflammatory microenvironment of CRLM-bearing liver tissue to donor liver.
Twenty-five patients undergoing resection for CRLM were recruited, 13 of whom developed intrahepatic recurrence within 18 months. Biopsies were obtained from tumour and normal liver tissue adjacent to and distal from, the tumour. Donor liver biopsies were obtained during transplantation. Biopsies were cultured and conditioned media (CM) screened for 102 inflammatory mediators. Twelve of these were validated by Luminex assay. Transwell assays measured cancer cell chemotaxis. Polymorphonuclear leukocytes (PMN) and lymphocytes were quantified in H&E sections.
Fewer periportal tissue-resident PMN were present in metastatic liver compared to donor liver. Patients with the fewest PMN in liver tissue distal to their tumour had a shorter time to intrahepatic recurrence (P < 0.001). IL-6, CXCL1, CXCL5, G-CSF, GM-CSF, VEGF, LIF, and CCL3 were higher in liver-bearing CRLM compared to donor tissue. Consequently, cancer cells migrated equally towards CM of all regions of metastatic liver but not towards donor liver CM.
The local inflammatory environment may affect both immune cell infiltration and cancer cell migration contributing to recurrence following resection for CRLM.
KeywordsLeukocytes Colorectal liver metastases Immune microenvironment Tumour progression
Chemokine (C–C motif) ligand
Colorectal liver metastases,
Chemokine (C-X-C motif) ligand
Leukemia inhibitory factor
Macrophage migration inhibitory factor
Vascular endothelial growth factor
FH: study design and conception, data collection, analysis and interpretation, drafted, reviewed, edited, and approved manuscript. CH: study design and conception, data analysis and interpretation, drafted, and approved manuscript. LAE: data collection and analysis, and reviewed and approved manuscript. FC: data collection and analysis, and reviewed and approved manuscript. AL: data analysis, and reviewed and approved manuscript. DM: provided clinical samples, and reviewed and approved manuscript. EH: provided clinical samples, and reviewed and approved manuscript. NN: data collection and analysis, and reviewed and approved manuscript. JGG: study design and conception, provided clinical samples, and reviewed and approved manuscript. EJR: study conception, design and supervision, data collection, and analysis, and drafted and approved manuscript. CO’F: study conception, design and supervision, and drafted and approved manuscript.
This work was supported by grants from the Health Research Board of Ireland (RP 2008/189) to C.O’Farrelly and HRA-POR-2013-281 to E.J. Ryan and a Science Foundation Ireland Investigator Award (132/IA/1667) to C. O’Farrelly.
Compliance with ethical standards
Conflict of interest
The authors declare they have no conflict of interest.
Ethical approval and ethical standards
All protocols were approved by St. Vincent’s University Hospital Ethics Committee in accordance with the ethical guidelines of the 1975 Declaration of Helsinki.
Informed consent was provided by all patients prior to undergoing resection. A patient information leaflet was provided and all queries addressed by F. Hand before consent was provided. Where donor liver was obtained, informed consent was given by the donor’s family prior to sampling. Donor families were approached by a dedicated transplant coordinator. All protocols and study information were provided to the family prior to consenting.
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