Inhibition of oncogenic epidermal growth factor receptor kinase triggers release of exosome-like extracellular vesicles and impacts their phosphoprotein and DNA content.
Cancer cells emit extracellular vesicles (EVs) containing unique molecular signatures. Here, we report that the oncogenic EGF receptor (EGFR) and its inhibitors reprogram phosphoproteomes and cargo of tumor cell-derived EVs. Thus, phosphorylated EGFR (P-EGFR) and several other receptor tyrosine kinases can be detected in EVs purified from plasma of tumor-bearing mice and from conditioned media of cultured cancer cells. Treatment of EGFR-driven tumor cells with second generation EGFR kinase inhibitors (EKIs), including CI-1033 and PF-00299804 but not with anti-EGFR antibody (Cetuximab) or etoposide, triggers a burst in emission of exosome-like EVs containing EGFR, P-EGFR, and genomic DNA (exo-gDNA). The EV release can be attenuated by treatment with inhibitors of exosome biogenesis (GW4869) and caspase pathways (ZVAD). The content of P-EGFR isoforms (Tyr-845, Tyr-1068, and Tyr-1173), ERK, and AKT varies between cells and their corresponding EVs and as a function of EKI treatment. Immunocapture experiments reveal the presence of EGFR and exo-gDNA within the same EV population following EKI treatment. These findings suggest that targeted agents may induce cancer cells to change the EV emission profiles reflective of drug-related therapeutic stress. We suggest that EV-based assays may serve as companion diagnostics for targeted anticancer agents.
From the Montreal Children's Hospital, Research Institute of the McGill University Health Centre, McGill University, Montreal, Quebec H4A 3J1.
Laura
L
Montermini
From the Montreal Children's Hospital, Research Institute of the McGill University Health Centre, McGill University, Montreal, Quebec H4A 3J1.
Brian
B
Meehan
From the Montreal Children's Hospital, Research Institute of the McGill University Health Centre, McGill University, Montreal, Quebec H4A 3J1.
Delphine
D
Garnier
From the Montreal Children's Hospital, Research Institute of the McGill University Health Centre, McGill University, Montreal, Quebec H4A 3J1.
Wan Jin
WJ
Lee
From the Montreal Children's Hospital, Research Institute of the McGill University Health Centre, McGill University, Montreal, Quebec H4A 3J1.
Tae Hoon
TH
Lee
the University of Toronto Health Network, Toronto, Ontario M5G 1L7, and.
Abhijit
A
Guha
the McMaster University, Hamilton, Ontario L8S 4K1, Canada.
Khalid
K
Al-Nedawi
From the Montreal Children's Hospital, Research Institute of the McGill University Health Centre, McGill University, Montreal, Quebec H4A 3J1, janusz.rak@mcgill.ca.
Janusz
J
Rak
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry; D015415:Biomarkers / Q000378:metabolism; D001932:Brain Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D000068818:Cetuximab / Q000737:chemistry; D017077:Culture Media, Conditioned / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D066246:ErbB Receptors / Q000378:metabolism*; D005047:Etoposide / Q000737:chemistry; D055354:Exosomes / Q000378:metabolism*; D000067128:Extracellular Vesicles / Q000378:metabolism*; D005910:Glioma / Q000378:metabolism; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D009025:Morpholines / Q000737:chemistry; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D040901:Proteomics; D052999:Quinazolinones / Q000737:chemistry; D014162:Transfection
D000970:Antineoplastic Agents; D015415:Biomarkers; D017077:Culture Media, Conditioned; D009025:Morpholines; D010750:Phosphoproteins; D052999:Quinazolinones; C525726:dacomitinib; D005047:Etoposide; D004247:DNA; C420268:Canertinib; C512478:EGFR protein, human; C541485:EGFR protein, mouse; D066246:ErbB Receptors; D000068818:Cetuximab
DNA; anticancer drug; cancer; caspase; enzyme inhibitor; epidermal growth factor receptor (EGFR); exosome (vesicle); phosphorylation; protein kinase