Centrosome amplification mediates small extracellular vesicle secretion via lysosome disruption

Sophie D Adams,Judit Csere,Gisela D’Angelo,Edward P Carter,Maryse Romao,Teresa Arnandis,Martin Dodel,Hemant M Kocher,Richard Grose,Graça Raposo,Faraz Mardakheh,Susana A Godinho

doi:10.1016/j.cub.2021.01.028

Abstract

SummaryBidirectional communication between cells and their surrounding environment is critical in both normal and pathological settings. Extracellular vesicles (EVs), which facilitate the horizontal transfer of molecules between cells, are recognized as an important constituent of cell-cell communication. In cancer, alterations in EV secretion contribute to the growth and metastasis of tumor cells. However, the mechanisms underlying these changes remain largely unknown. Here, we show that centrosome amplification is associated with and sufficient to promote small extracellular vesicle (SEV) secretion in pancreatic cancer cells. This is a direct result of lysosomal dysfunction, caused by increased reactive oxygen species (ROS) downstream of extra centrosomes. We propose that defects in lysosome function could promote multivesicular body fusion with the plasma membrane, thereby enhancing SEV secretion. Furthermore, we find that SEVs secreted in response to amplified centrosomes are functionally distinct and activate pancreatic stellate cells (PSCs). These activated PSCs promote the invasion of pancreatic cancer cells in heterotypic 3D cultures. We propose that SEVs secreted by cancer cells with amplified centrosomes influence the bidirectional communication between the tumor cells and the surrounding stroma to promote malignancy.

Highlights

Centrosome amplification induces secretion of sEVs Our previous work demonstrated that centrosome amplification leads to proteomic changes in the secretome, including an increase in proteins associated with Extracellular vesicles (EVs), suggesting higher EV secretion in cells with amplified centrosomes.[10]
We observed that cell lines with higher levels of centrosome amplification secreted increased numbers of EVs, in particular small extracellular vesicle (SEV), demonstrating a significant correlation between extra centrosomes and SEV secretion (Figures 1A–1C and S1F)
Depletion of SAS-6, a protein important for centrosome duplication,[27] in cells exposed to DOX and Polo-like kinase 4 (PLK4) overexpression prevented both centrosome amplification and increased SEV secretion but had no effect on SEV secretion in control cells, suggesting that SEV secretion is a consequence of centrosomal alterations (Figures 1D and S1G–S1I)

Summary

Introduction

A variety of human cancer types often exhibit defects in the structure and number of centrosomes, the main microtubule organizing centers in animal cells.[1,2] Work in fly and mouse models has shown that centrosome abnormalities, in particular centrosome amplification, are not mere byproducts of tumorigenesis but rather play direct roles in promoting and accelerating tumor progression.[3,4,5,6] the full extent by which centrosome abnormalities contribute to tumorigenesis is still unclear, centrosome amplification can directly promote aneuploidy and cell invasion, which play important roles in malignant progression.[7,8,9] Recently, we reported that centrosome amplification induces the secretion of several proteins with pro-invasive properties, e.g., interleukin-8, which induces invasive behavior in neighboring cells.[10] This altered secretion is partially due to a stress response that results from increased reactive oxygen species (ROS) downstream of centrosome amplification.[10] the presence of amplified centrosomes can influence tumors in a non-cell-autonomous manner, via protein secretion, suggesting a broader and more complex role for these abnormalities in cancer

Methods

Results

Discussion

Conclusion