Phosphatase Shp2 regulates biogenesis of small extracellular vesicles by dephosphorylating Syntenin

Abstract

As novel mediators of cell‐to‐cell signalling, small extracellular vesicles (sEVs) play a critical role in physiological and pathophysiological processes. To date, the molecular mechanisms that support sEV generation are incompletely understood. Many kinases are reported for their roles in sEV generation or composition, whereas the involvement of phosphatases remains largely unexplored. Here we reveal that pharmacological inhibition and shRNA‐mediated down‐regulation of tyrosine phosphatase Shp2 significantly increases the formation of sEVs. By Co‐immunoprecipitation (Co‐IP) and in vitro dephosphorylation assays, we identified that Shp2 negatively controlled sEV biogenesis by directly dephosphorylating tyrosine 46 of Syntenin, which has been reported as a molecular switch in sEV biogenesis. More importantly, Shp2 dysfunction led to enhanced epithelial sEV generation in vitro and in vivo. The increase of epithelial sEVs caused by shRNA‐mediated down‐regulation of Shp2 promoted macrophage activation, resulting in strengthened inflammation. Our findings highlight the role of Shp2 in regulating sEV‐mediated epithelial‐macrophage crosstalk by controlling sEV biogenesis through dephosphorylation of Syntenin Y46. The present study determines the strengthened inflammatory characteristics of alveolar macrophages elicited by epithelial sEVs transferred intercellularly. These findings provide a basis for understanding the mechanism of sEV formation and relevant function in epithelial‐macrophage crosstalk.

Targeting protein phosphatases for the treatment of inflammation-related diseases: From signaling to therapy

“… 34 SHP2 is a key regulator of immune cell-mediated inflammation through its direct influence on the immune response or the inflammation process. 30 The functions of SHP2 in inflammatory diseases have been widely studied in inflammatory bowel disease (IBD), 37 neuroinflammation, 38 , 39 autoimmune diseases 40 and lung inflammation 41 as we next discussed (Fig. 6 ).…”

“… 58 SHP2 was also shown to regulate the biogenesis of small extracellular vesicles by dephosphorylating its substrate syntenin Tyr46, while the knockdown of SHP2 promoted macrophage activation and lung inflammation. 41 Increased expression of SHP2 in endothelial cells contributed to radiation-induced endothelial dysfunction and the subsequent establishment of an inflammatory microenvironment during radiation-induced lung injury. 59 Therefore, most of the current evidence supports a proinflammatory role for SHP2 in asthma and lung inflammation.…”

Tumor-Derived Extracellular Vesicles Regulate Cancer Progression in the Tumor Microenvironment

“…Syndecan-syntenin complexes bind ESCRT-I and ESCRT-III via Alix, leading to enhanced ILVs biogenesis ( Baietti et al, 2012 ). Moreover, tyrosine phosphatase Shp2 has been found to inhibit exosomes biogenesis via dephosphorylating syntenin ( Zhang Y. et al, 2021 ). CD63, belonging to tetraspanin family, is associated with sorting cargoes into exosomes ( Theos et al, 2006 ; van Niel et al, 2011 ).…”

An Emerging Frontier in Intercellular Communication: Extracellular Vesicles in Regeneration

“…Notwithstanding these studies, a comprehensive understanding of this phenomenon across tissues and organisms is lacking, and there is limited understanding of mechanisms by which EV output is controlled. Phosphorylation of EV biogenesis regulators is one possible mechanism: pyruvate kinase type M2 can promote EV release through phosphorylation of SNAP-23, while the phosphatase Shp2 inhibits EV release through dephosphorylation of Syntenin, but it is not clear whether these mechanisms are relevant during regeneration ( Wei et al, 2017 ; Zhang et al, 2021b ). Although continued development of methods to isolate EVs and monitor their local production in regions of tissue damage in vivo will be needed ( Brock et al, 2019 ; Verweij et al, 2019 ), investigations in emerging models will expand our understanding of how injury is transduced into changes in EV output.…”