Wnt Activity and Cell Proliferation Are Coupled to Extracellular Vesicle Release in Multiple Organoid Models

Abstract

Extracellular vesicles (EV) are considered as a potential tool for early disease diagnosis; however, factors modifying EV release remain partially unknown. By using patient-derived organoids that capture the cellular heterogeneity of epithelial tissues, here we studied the connection between the Wnt-producing microniche and EV secretion in multiple tissues. Although nearly all cells in pancreatic ductal (PD) and pancreatic ductal adenocarcinoma (PDAC) samples expressed porcupine (PORCN), an enzyme critical for Wnt secretion, only a subpopulation of lung bronchiolar (NL) and lung adenocarcinoma (LUAD) organoid cells produced active Wnt. The microniche for proliferating cells was shaped not only by PORCN + cells in NL and LUAD organoids but also by fibroblast-derived EVs. This effect could be blocked by using Wnt secretion inhibitors. Whereas inhibiting Wnt secretion in PD NL or LUAD organoids critically changed both cell proliferation and EV release, these were uncoupled from each other in PDAC. Sorting for CD133 identified a cell population in the LUAD microniche that produced organoids with a high percentage of PORCN + and proliferating cells and an elevated EV secretion, which may explain that CD133 marks LUAD cells with malignant behavior. Collectively, we show here that high cell proliferation rate, induced by Wnt pathway activation, is coupled to a higher EV release, a critical finding that may be considered when developing EV-based diagnostic tools.

Differential Transcriptomic Profiles Following Stimulation with Lipopolysaccharide in Intestinal Organoids from Dogs with Inflammatory Bowel Disease and Intestinal Mast Cell Tumor

“…Using canine-specific LGR5 probes, we previously detected ISCs within the intestine organoid crypts [ 30 ], demonstrating that proliferation is restricted to crypt-like budding structures [ 64 ]. Ki67 has been used as a proliferation marker as its expression is strongly associated with proliferation and growth [ 65 , 66 ]. Other intestinal stem cell markers and genes associated with proliferation, notably proliferating cell nuclear antigen ( PCNA ), prominin 1 ( PROM1 ), HOP homeobox ( HOPX ), and olfactomedin 4 ( OLFM4 ), were also increased in LPS-treated IBD intestinal organoids and tumor enteroids.…”

Advancement of Organoid Technology in Regenerative Medicine

“…Finally, the effect of genetics in the production of EVs is suggested by studying the role of Apc mutation in activation of the Wnt pathway and EV production as the downstream result [ 122 ]. Similarly, in a study by Sándor et al on lung adenocarcinoma organoids, the role of the Wnt pathway on cell proliferation and EV secretion has been indicated [ 124 ]. In a recent study by Taha et al, it has been indicated that matrix metalloproteinase 3 (MMP3) has a crucial role in maintaining the physical integrities of EVs and any reduction in MMP3 results in disorganization of EV structures, inhibition of their protumorigenic properties, and finally a reduction in tumor proliferation and progression [ 116 ].…”

Lung Cancer Organoids: The Rough Path to Personalized Medicine

“…Sàndor and coworkers used LUAD organoids to identify a Wnt-producing microniche composed by both cancer cells and fibroblasts, which was able to influence cancer cell proliferation and extracellular vesicle release. The same authors identified a CD133high subpopulation in LCOs with stem cell characteristics, such as elevated self-renewal and Wnt activity or responsiveness [ 56 ]. Finally, Ma and coworkers analyzed the gene expression profiles of LCOs derived from LUAD and LUSC, finding a differential regulation of pathways involved in immune regulation, inflammation, MAPK, and Rap1 activation between the two histotypes [ 57 ].…”