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Extracellular vesicles (EVs) encompass a diverse set of membrane-derived particles released from cells and are found in numerous biological matrices and the extracellular space. Specific classes of EVs include apoptotic bodies, exosomes, and microvesicles, which vary in their size, origin, membrane protein expression, and interior cargo. EVs provide a mechanism for shuttling cargo between cells, which can influence cell physiology by transporting proteins, DNA, and RNA. EVs are an abundant component of the tumor microenvironment (TME) and are proposed to drive tumor growth and progression by communicating between fibroblasts, macrophages, and tumor cells in the TME. The cargo, source, and type of EV influences the pro- or anti-tumoral role of these molecules. Therefore, robust EV isolation and characterization techniques are required to ensure accurate elucidation of their association with disease. Here, we summarize different EV subclasses, methods for EV isolation and characterization, and a selection of current clinical trials studying EVs. We also review key studies exploring the role and impact of EVs in the TME, including how EVs mediate intercellular communication, drive cancer progression, and remodel the TME.

Triphysaria is a facultative parasitic plant in the Orobanchaceae that parasitizes the roots of a wide range of host plants including Arabidopsis, Medicago, rice and maize. The important exception to this broad host range is that Triphysaria rarely parasitize other Triphysaria. We explored self and kin recognition in Triphysaria versicolor and showed that exudates collected from roots of host species, Arabidopsis thaliana and Medicago truncatula, induced haustorium development when applied to the roots of Triphysaria seedlings in vitro while those collected from Triphysaria did not. In mixed exudate experiments, Triphysaria exudates did not inhibit the haustorium-inducing activity of those from host roots. Interestingly, when roots of Triphysaria seedlings were treated with either horseradish peroxidase or fungal laccase, the extracts showed haustorium-inducing factor (HIF) activity, suggesting that Triphysaria roots contain the proper substrates for producing HIFs. Transgenic Triphysaria roots overexpressing a fungal laccase gene TvLCC1 showed an increased responsiveness to a known HIF, 2,6-dimethoxy benzoquinone (DMBQ), in developing haustoria. Our results indicate kin recognition in Triphysaria is associated with the lack of active HIFs in root exudates. Treatment of Triphysaria roots with enzymatic oxidases activates or releases molecules that are HIFs. This study shows that exogenously applied oxidases can activate HIFs in Triphysaria roots that had no previous HIF activity. Further studies are necessary to determine if differential oxidase activities in host and parasite roots account for the kin recognition in haustorium development.