Immunomagnetic isolation of circulating melanoma cells and detection of PD-L1 status

PLoS One. 2019 Feb 8;14(2):e0211866. doi: 10.1371/journal.pone.0211866. eCollection 2019.

Abstract

Personalised medicine targeted to specific biomarkers such as BRAF and c-Kit has radically improved the success of melanoma therapy. More recently, further advances have been made using therapies targeting the immune response. In particular, therapies targeting the PD-1/PD-L1 or CTLA-4 axes alone or in combination have shown more sustained responses in 30-60% of patients. However, these therapies are associated with considerable toxicities and useful biomarkers to predict responders and non-responders are slow to emerge. Here we developed a reliable melanoma circulating tumor cell (CTC) detection method with PD-L1 evaluation on CTCs. A set of melanoma cell surface markers was tested as candidates for targeted melanoma CTC isolation and a melanoma specific immunostaining-based CTC identification protocol combined with PD-L1 detection was established. In vitro testing of the effect of exposure to blood cells on melanoma cell PD-L1 expression was undertaken. Immunomagnetic targeting isolated melanoma CTCs in up to 87.5% of stage IV melanoma patient blood samples and 3 8.6% of these had some PD-L1 expressing CTCs. Our in vitro data demonstrate PD-L1 induction on melanoma cells in the blood.This study established a robust, reliable method to isolate melanoma CTCs and detect expression of PD-L1 on these cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • B7-H1 Antigen / blood
  • B7-H1 Antigen / genetics*
  • Biomarkers, Tumor / blood
  • Biomarkers, Tumor / genetics*
  • CTLA-4 Antigen / blood
  • CTLA-4 Antigen / genetics
  • Female
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Immunomagnetic Separation / methods*
  • Immunophenotyping
  • Lung Neoplasms / blood
  • Lung Neoplasms / diagnosis*
  • Lung Neoplasms / genetics
  • Lung Neoplasms / secondary
  • Male
  • Melanoma / blood
  • Melanoma / diagnosis*
  • Melanoma / genetics
  • Melanoma / secondary
  • Middle Aged
  • Neoplasm Staging
  • Neoplastic Cells, Circulating / immunology
  • Neoplastic Cells, Circulating / pathology
  • Programmed Cell Death 1 Receptor / blood
  • Programmed Cell Death 1 Receptor / genetics
  • Skin Neoplasms / blood
  • Skin Neoplasms / diagnosis*
  • Skin Neoplasms / genetics
  • Skin Neoplasms / pathology

Substances

  • B7-H1 Antigen
  • Biomarkers, Tumor
  • CD274 protein, human
  • CTLA-4 Antigen
  • CTLA4 protein, human
  • PDCD1 protein, human
  • Programmed Cell Death 1 Receptor

Grants and funding

JP is recipient of a Rotary Health Australia PhD Scholarship. DB, PdS, YM and TB are supported by the Cancer Institute New South Wales through the Centre for Oncology Education and Research Translation (CONCERT).