Naive primary neutrophils play a dual role in the tumor microenvironment
Kehinde Adebayo Babatunde,
Rupsa Datta,
Nathan W. Hendrikse,
Jose M. Ayuso,
Anna Huttenlocher,
Melissa C. Skala,
David J. Beebe,
Sheena C. Kerr
Affiliations
Kehinde Adebayo Babatunde
Department of Pathology & Laboratory Medicine, University of Wisconsin, Madison, WI 53705, USA
Rupsa Datta
Morgridge Institute for Research, Madison, WI 53715, USA
Nathan W. Hendrikse
Department of Pathology & Laboratory Medicine, University of Wisconsin, Madison, WI 53705, USA
Jose M. Ayuso
Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin, Madison, WI 53705, USA; Department of Dermatology, University of Wisconsin, Madison, WI 53705, USA
Anna Huttenlocher
Departments of Pediatrics and Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53792, USA
Melissa C. Skala
Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin, Madison, WI 53705, USA
David J. Beebe
Department of Pathology & Laboratory Medicine, University of Wisconsin, Madison, WI 53705, USA; Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin, Madison, WI 53705, USA
Sheena C. Kerr
Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin, Madison, WI 53705, USA; Corresponding author
Summary: The tumor microenvironment (TME) is characterized by a network of cancer cells, recruited immune cells, and extracellular matrix (ECM). However, the specific role of neutrophils during tumor development, and their interactions with other immune cells is still not well understood. Here, we use both standard well plate culture and an under oil microfluidic (UOM) assay with an integrated ECM bridge to elucidate how naive primary neutrophils respond to tumor cells. Our data demonstrated that tumor cells trigger cluster formation in neutrophils accompanied with the generation of reactive oxygen species (ROS) and neutrophil extracellular trap (NET) release. Using label-free optical metabolic imaging (OMI), we observed changes in the metabolic activities of primary neutrophils during the different clustering phases when challenged with tumor cells. Finally, our data demonstrates that neutrophils in direct contact, or in close proximity, with tumor cells exhibit greater metabolic activities compared to non-contact neutrophils.
