HAS3-induced extracellular vesicles from melanoma cells stimulate IHH mediated c-Myc upregulation via the hedgehog signaling pathway in target cells

Abstract

Intercellular communication is fundamental to the survival and maintenance of all multicellular systems, whereas dysregulation of communication pathways can drive cancer progression. Extracellular vesicles (EVs) are mediators of cell-to-cell communication that regulate a variety of cellular processes involved in tumor progression. Overexpression of a specific plasma membrane enzyme, hyaluronan synthase 3 (HAS3), is one of the factors that can induce EV shedding. HAS3, and particularly its product hyaluronan (HA), are carried by EVs and are known to be associated with the tumorigenic properties of cancer cells. To elucidate the specific effects of cancerous, HAS3-induced EVs on target cells, normal human keratinocytes and melanoma cells were treated with EVs derived from GFP-HAS3 expressing metastatic melanoma cells. We found that the HA receptor CD44 participated in the regulation of EV binding to target cells. Furthermore, GFP-HAS3positive EVs induced HA secretion, proliferation and invasion of target cells. Our results suggest that HAS3-EVs contains increased quantities of IHH, which activates the target cell hedgehog signaling cascade and leads to the activation of c-Myc and regulation of claspin expression. This signaling of IHH in HAS3-EVs resulted in increased cell proliferation. Claspin immunostaining correlated with HA content in human cutaneous melanocytic lesions, supporting our in vitro findings and suggesting a reciprocal regulation between claspin expression and HA synthesis. This study shows for the first time that EVs originating from HAS3 overexpressing cells carry mitogenic signals that induce proliferation and epithelial-to-mesenchymal transition in target cells. The study also identifies a novel feedback regulation between the hedgehog signaling pathway and HA metabolism in melanoma, mediated by EVs carrying HA and IHH.

Inflammation, Extracellular Matrix Remodeling, and Proteostasis in Tumor Microenvironment

“…The role of HAS3 in tumor progression is not related to its enzymatic activity, eventually altered by post-translational modifications, but more to its involvement in the induction of extracellular vesicles (EVs) shedding. Arasu et al observed that increased amounts of EVs derived from GFP-HAS3 expressing metastatic melanoma cells induced HA secretion, proliferation, and invasion of target keratinocytes and primary melanoma cell line [ 64 ]. As described here, these HAS3-rich EVs contained high amounts of HA and, importantly, CD44 that participated in the regulation of EV binding to target cells [ 64 ].…”

“…Arasu et al observed that increased amounts of EVs derived from GFP-HAS3 expressing metastatic melanoma cells induced HA secretion, proliferation, and invasion of target keratinocytes and primary melanoma cell line [ 64 ]. As described here, these HAS3-rich EVs contained high amounts of HA and, importantly, CD44 that participated in the regulation of EV binding to target cells [ 64 ]. In addition to cell functions’ associated genes induction, target cells may undergo EMT.…”

Balance Between Tooth Size and Tooth Number Is Controlled by Hyaluronan

“…Regarding its signaling properties, it has been shown that HA plays an important role during embryonic development regulating cell migration, shape, and proliferation ( Spicer and Tien, 2004 ). HAS3 is the most active enzyme of the HAS family and polymerizes shorter HA chains ( Itano et al, 1999 ; Itano and Kimata, 2002 ) that stimulates cell signaling, proliferation, and epithelial-to-mesenchymal transition in cancer models ( Kultti et al, 2014 ; Kuo et al, 2017 ; Arasu et al, 2019 ). HAS3 expression is higher in the dental epithelium than the mesenchyme, with the distribution of CD44 appearing complementary to HAS3 in the dental epithelium.…”

Bacterial Membrane Vesicles in Pneumonia: From Mediators of Virulence to Innovative Vaccine Candidates

“…(O)MVs bind to the bronchial epithelium, as shown for P. aeruginosa OMVs [ 69 ], and alveolar epithelial cells, as demonstrated for A. baumannii OMVs and S. pneumoniae MVs [ 51 , 70 ]. Notably, for EVs an important role of CD44 in the EV binding process has been well documented [ 71 , 72 ]. As CD44 is the hyaluronic receptor, and hyaluronan forms an important constituent of both the alveolar epithelial glycocalyx and airway mucus, a similar “attachment chemistry” may also exist in (O)MVs but remains to be identified [ 73 , 74 ].…”