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Characterizing human head and neck tumors and cancer cell lines with mass cytometry

Tess M. Brodie, Vinko Tosevski, Michaela Medová Cytometry PART A, Volume 93, Issue 4, 406-410 (2018)

PURPOSE: A 42‐parameter panel (34 antibody targets, cisplatin for live/dead, iridium for cell identification, and 6 barcodes) was developed for the characterization of cell types and signaling pathways present in tumor tissues or cell lines originating from primary, metastatic and recurrent head and neck squamous cell carcinoma (HNSCC), the most prevalent malignant tumor of the head and neck region and the 6th most common form of non‐skin cancer worldwide 1 (Table 1). Importantly, due to overlap in targets and markers across diverse tumor entities [e.g., HER2 amplification is present in both breast and gastric tumors 2, 3], the described panel may be appropriate for use in preclinical models of additional distinct solid as well as hematological malignancies. This panel has been successfully tested on fresh, formalin‐fixed, and frozen cells originating from cell lines derived from primary and metastatic/recurrent tumors and from patients' tumor tissue.

CELL TYPE: Cancer cell lines, HNSCC Tumors

MACHINE: Fluidigm CyTOF 2.1 Helios

Marker Target Clone Fluorochrome Purpose
CD45 Human HI30 Purified Leukocytes
CD3 Human SK7 Purified T cells
CD31 Human 390 Purified Endothelial cells
CD10 Human HI10a Purified Cancer associated fibroblasts
Vimentin Human D21H3 Purified Mesenchymal cells
CD68 Human Y1/82A Purified Macrophages and DCs
CD24 Human ML5 Purified B cells
CD324 Human 24E11 Purified Epithelial cells
PD-1 Human EH12.2H7 Purified Tolerance promotion
CD274 Human 29E.2A3 Purified Tolerance promotion
CD28 Human CD28.2 Purified T cell activation
Ki-67 Human B56 Purified Proliferation
p53 Human 261352 Purified Tumor suppressor protein
PARP, cleaved Human F21-852 Purified Repair of single stranded DNA
ERK1/2 (pT202/pY204) Human D13.14.4E Purified Proliferation
EGFR (pY845) Human D7A5 Purified Proliferation
HER2/ErbB2 Human 29D8 Purified Proliferation
Akt (pS473) Human D9E Purified Survival
Met (pY1234/pY1235) Human Purified Proliferation
DDR2 Human 290804 Purified Proliferation
TRAF3 Human B1-6 Purified Associated with HNSCC
EGF Receptor Human Purified Proliferation
YAP1 Human 867711 Purified Proliferation
Hes1 Human 4H1HES1 Purified Transcription suppressor
EphA2 Human SHM16 Purified Migration and proliferation
S6 (pS235/pS236) Human N7-548 Purified Increased translation
c-Met Human eBioclone 97 Purified Proliferation
CD29 Human TS2/16 Purified Metastasis
MMP-9 Human 4H3 Purified Metastasis
TIMP-3 Human 277128 Purified Metastasis inhibitor
MDR1 Human UIC2 Purified Drug resistance
ALDH1A1 Human 703410 Purified Associated with cancer stem cells
CD166 Human 3A6 Purified Associated with cancer stem cells
CD44 Human Purified Associated with cancer stem cells