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Exosome binding/uptake mediated by Integrin
Exosomes are a type of extracellular vesicle (EV) of endocytic origin that are released by virtually all cells in multicellular organisms and carry out important intercellular communication functions through the transfer of their biomolecular cargo, which includes lipids, proteins, nucleic acids and metabolites, between the producing and the recipient/target cells . Exosomes produced by cancer cells have been shown to influence many processes related to cancer progression and metastasis, such as tumor cell proliferation and invasion, angiogenesis, tumor microenvironment promotion and remodelling, chemotherapy resistance, and immune suppression (recently reviewed in [2,3,4]).
Colorectal cancer (CRC) has a high incidence and is a major cause of cancer-related mortality worldwide, with approximately 25% of patients developing peritoneal metastasis, a condition associated with a bleak prognosis as current treatments are less effective in these patients . The CRC peritoneal dissemination cascade involves sequential shedding of cancer cells from the primary colonic or rectal tumor, their transport through the peritoneal cavity following the physiological flow of peritoneal fluid, their adhesion to the peritoneal mesothelial cells (PMCs) that form a barrier lining all peritoneal organs, and the subsequent invasion of cancer cells through this mesothelial cell barrier and the underlying stroma to establish new metastatic foci in the peritoneum, omentum and bowel serosa. In addition to acting as a barrier, PMCs also play an active role in the pathogenesis of peritoneal metastasis by differentiating into cancer-associated fibroblasts (CAFs) through a process termed Mesothelial-to-Mesenchymal Transition (MMT), which fuels peritoneal metastasis and immune evasion (reviewed in ).
In epithelial ovarian cancer, which predominantly disseminates via peritoneal metastasis, exosomes produced by ovarian cancer cells favor different steps of the peritoneal dissemination cascade and have therefore been recognized to play crucial roles in the pathogenesis of the disease . Of particular relevance, cancer-derived exosomes in malignant ascites interact with cancer cells promoting their survival, migration and invasion capacities, and with PMCs inducing their apoptosis, disrupting the mesothelial barrier and reprogramming them into CAFs through MMT. For CRC, however, current knowledge about the roles played by exosomes in the pathogenesis and peritoneal metastasis cascade is sparse. At the same time, the molecules that mediate the interactions between cancer-derived exosomes and their target PMCs and tumor cells mostly remain obscure.
Previously we employed the human colorectal adenocarcinoma Colo-320 cell line  as a useful model to study cancer-related processes such as tumorigenesis, metastasis, and tumor cell adhesion, as well as the functions and interplay between tetraspanins and cell adhesion molecules . Colo-320 cells possess the distinctive feature of lacking endogenous expression of tetraspanin CD9 while they abundantly express integrin α5β1 but not many other members of the β1 integrin subfamily. CD9 is a widely distributed tetraspanin that associates with other transmembrane proteins, including integrins and ADAM metalloproteases in TEMs (Tetraspanin-Enriched Microdomains) (reviewed in ). CD9 is involved in cell adhesion, motility, sperm–egg fusion, tumorigenesis and metastasis and an inverse correlation between expression of CD9 and metastatic potential and patient survival rate has been established in many types of cancer, including CRC (reviewed in ).
Several reports indicate that integrin α5β1 can engage in cis (on the same cell) and trans (on different cells) interactions with the distintegrin (Dis) domain of the metalloproteinase ADAM17/TACE . Recent study showed that the tetraspanin CD9 negatively regulated integrin α5β1-mediated adhesion of cancer cells both to its canonical ligand fibronectin and to ADAM17 as a novel ligand Furthermore, it has reported that CD9 interacts directly with ADAM17 on the cell surface and regulates negatively both the sheddase and adhesive activities of this metalloproteinase . Therefore, we decided to investigate whether integrin α5β1 and ADAM17 interaction and the regulatory effects of CD9 also played a role in mediating the binding of exosomes and their uptake by cancer or mesothelial recipient cells, which could bear relevance during the peritoneal dissemination of colorectal carcinomas.
We report here that the interaction between integrin α5β1 on colorectal carcinoma cells and on PMCs and its ligand ADAM17 on exosomes mediates the binding and uptake of cancer-derived exosomes. Furthermore, this process was negatively regulated by the expression of tetraspanin CD9 on the surface of exosomes.
2.1. Characterization of EVs Derived from Colo-320, Colo-320/CD9 and Colo-320/ADAM17-KO Human Adenocarcinoma Cells
2.2. Fibronectin Is Not the Ligand of Integrin α5β1 That Mediates Interactions of Exosomes with Colo-320 Cancer Cells
2.3. Expression of CD9 Reduces Interactions between Cancer Cells and Exosomes Mediated by Cellular Integrin α5β1 and Exosomal ADAM17
2.4. Uptake of Exosomes by Colo-320 Cancer Cells Depends on Exosomal ADAM17 and Is Inhibited by CD9
This entry is adapted from 10.3390/ijms22189938
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