Cytokine Profiling in Low- and High-Density Small Extracellular Vesicles from Epidermoid Carcinoma Cells

Joseph P Flemming; Brianna L Hill; Lauren Anderson-Pullinger; Larry A Harshyne; Mỹ G Mahoney

doi:10.1016/j.xjidi.2021.100053

Cytokine Profiling in Low- and High-Density Small Extracellular Vesicles from Epidermoid Carcinoma Cells

JID Innov. 2021 Aug 24;1(4):100053. doi: 10.1016/j.xjidi.2021.100053. eCollection 2021 Dec.

Authors

Joseph P Flemming¹, Brianna L Hill¹, Lauren Anderson-Pullinger², Larry A Harshyne², Mỹ G Mahoney^{1

3}

Affiliations

¹ Department of Dermatology & Cutaneous Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
² Department of Medical Oncology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
³ Department of Otolaryngology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.

Abstract

Exosomes or small extracellular vesicles (sEVs) are membrane-bound nanoparticles that carry various macromolecules and act as autocrine and paracrine signaling messengers. In this study, sEVs from epidermoid carcinoma cells influenced by membrane presentation of the glycoprotein desmoglein 2 and its palmitoylation state were investigated. In this study, sEVs were isolated by sequential ultracentrifugation followed by iodixanol density gradient separation. They were then subjected to multiplex profiling of cytokines associated with the surface of intact sEVs. The results revealed a previously undescribed active sorting of cytokines onto the surface of low-density and high-density sEV subpopulations. Specifically, an altered surface presentation of desmoglein 2 decreased FGF-2 and VEGF in low-density sEVs. In addition, in response to desmoglein 2, IL-8 and RANTES were increased in low-density sEVs but only slightly decreased in high-density sEVs. Finally, IL-6 and G-CSF were increased dramatically in high-density sEVs. This comprehensive analysis of the cytokine production profile by squamous cell carcinoma‒derived sEVs highlights their contribution to immune evasion, pro-oncogenic and proangiogenic activity, and the potential to identify diagnostic disease biomarkers.

Keywords: Dsg2, desmoglein 2; ED, extracellular domain; EV, extracellular vesicle; HD-sEV, high-density small extracellular vesicle; LD-sEV, low-density small extracellular vesicle; SCC, squamous cell carcinoma; sEV, small extracellular vesicle.