bioRxiv | 2019
Differential in vivo biodistribution of 131 I-labeled exosomes from diverse cellular origins and its implication in the theranostic application
Abstract
Exosomes are critical mediators of intercellular crosstalk and regulator of cellular/tumor microenvironment. Exosomes have great prospects for clinical application as theranostic and prognostic probe. Nevertheless, the advancement of the exosomes research has been thwarted by limited knowledge elucidating the most efficient isolation method and their in vivo trafficking. Here we have showed that combination of two size-based methods using 0.20 µm syringe filter and 100k centrifuge membrane filter followed by ultracentrifugation method yields a greater number of uniform exosomes. We also demonstrated the visual representation and quantification of differential in vivo distribution of radioisotope 131I-labelled exosomes from diverse cellular origins, e.g., tumor cells with or without treatments (HET0016 and GW2580), myeloid-derived suppressor cells and endothelial progenitor cells. We also determined that the distribution was dependent on the protein/cytokine contents of the exosomes. The applied in vivo imaging modalities can be utilized to monitor disease progression, metastasis, and exosome-based targeted therapy. Abbreviations bFGF basic fibroblast growth factor CSF1R colony stimulating factor 1 receptor CT computed tomography CTLA4 cytotoxic T-lymphocyte-associated protein 4 EGF epidermal growth factor EMT epithelial to mesenchymal transition EVs extracellular vesicles EPCs endothelial progenitor cells FasL Fas ligand G-CSF granulocyte-colony stimulating factor GM-CSF granulocyte-macrophage colony-stimulating factor HGF hepatocyte growth factor HSP heat shock protein ICAM-1 intercellular adhesion molecule 1 IFN-gamma interferon gamma IL – 1beta interleukin-1 beta IL – 1ra interleukin-1 receptor antagonist IL – 2 interleukin-2 IL – 4 interleukin-4 IL – 6 interleukin-6 IL – 7 interleukin-7 IL – 10 interleukin-10 IL – 12 interleukin-12 IL – 13 interleukin-13 IL – 17 interleukin-17 KC keratinocyte-derived chemokine LIX lipopolysaccharide-induced CXC chemokine M-CSF macrophage colony-stimulating factor MCP-1 monocyte chemoattractant protein 1 MDC macrophage-derived chemokine MDSCs myeloid derived suppressor cells MFP mammary fat pad MIP-1α macrophage-inflammatory protein-1alpha MMP-2 matrix metalloproteinase-2 MRI magnetic resonance imaging NIS sodium iodide symporter NTA nanoparticle tracking analysis PET positron emission tomography PF-4 platelet factor 4 RANTES regulated on activation, normal T cell expressed and secreted ROIs region of interest SDF-1α stromal cell-derived factor-1 SEM standard error of the mean SPECT single-photon emission computed tomography SCF stem cell factor TAMs tumor-associated macrophages TEM transmission electron microscopy TIMP 2 tissue inhibitors of metalloproteinases 2 TLPC thin layer paper chromatography TME tumor microenvironment TNF-α tumor necrosis factor-α TSLP thymic stromal lymphopoietin UC ultracentrifugation VEGF-A vascular endothelial growth factor A VEGFR2 vascular endothelial growth factor receptor 2.