The cellular transfer of the tumor suppressor p53 via extracellular vesicles confers p53-activities to target cells

Background

The tumor suppressor TP53 gene is mutated in over 96% of cases in high-grade serous ovarian cancers (HGSOC), the most common subtype of ovarian cancer (OC). OC is one of the leading causes of death in patients with gynecological malignancies and the tumor secretome has been reported to play a central role in tumorigenesis. TP53 deficiency or mutations shift the cellular secretome from a wild-type-driven tumor-suppressive to a tumor-promoting secretome. Extracellular vesicles (EVs) as part of the secretome are recently recognized as novel mediators within the OC microenvironment. The contribution of p53 to the biogenesis and cargo sorting of OC cell-derived EVs and functional implications for intercellular communication remain unknown.  

 

Methods

OC cell lines with defined p53 status were designed and analysed by transcriptomics and proteomics of the cells and their derived EVs, both. Gene expression analysis of fibroblasts were conducted upon treatment with EVs. High-resolution microscopy was applied to detect the enrichment of p53 and its transfer via EVs to recipient cells. 

 

Results

p53-wild-type cells (WTp53) released more EVs and these EVs had a distinct protein cargo as compared to EVs from knock-out (KOp53) or mutated p53 (mutp53). Consequently, WTp53-EVs induced an NF-κB and STAT3-mediated pro-inflammatory phenotype (IL8, IL6, CXCL1) in fibroblasts used as target cells. In contrast, EVs derived from KOp53 cells induced the expression of TGFß targets such as extracellular matrix molecules (COMP, FN1, CCN2). Unexpectedly, p53, but not mutp53 (R175H or R273H), was enriched in p53-EVs and transferred to recipient cells as observed using high-resolution microscopy. Moreover, the EV-mediated transfer of p53 to p53-deficient OC cells rescued resistance to p53-induced apoptosis

 

Conclusion

These findings may open the avenue to explore WTp53-EVs as therapeutic tools to reactivate or replace the function of mutated or inactive p53 in tumor cells and the microenvironment.

 

Keywords

TP53, Ovarian cancer, Extracellular vesicles, Fibroblasts.

 

Funding/Acknowledgments

This work was supported by grants from Deutsche Forschungsgemeinschaft (KFO325, project 329116008 and GRK2573, project 416910386 to EPvS).  

 

Authors

Manuel Linder1, Bilal Alashkar Alhamwe1, Viviane Ponath1, Florian Finkernagel2, Rambod Nikbakhsh1, Andrea Nist2, Julia Teply-Szymanski2, Thorsten Stiewe2, Witold Szymański3, Vanessa Beutgen3, Johannes Graumann3, Christian Preußer4, Ralf Jacob5, Elke Pogge von Standmann1, 4 (Corresponding Author: poggevon[at]staff.uni-marburg[dot]de)

 

1 Institute for Tumor Immunology, Philipps-University, 35043, Marburg, Germany and Core Facility Extracellular Vesicles, Philipps-University, 35043, Marburg, Germany
2 Institute of Molecular Oncology and Genomics Core Facility, Member of the German Center for Lung Research (DZL), Philipps-University, 35043 Marburg, Germany; Institute of Lung Health, Justus Liebig University, 35392 Giessen, Germany

3 Institute of Translational Proteomics & Core Facility Translational Proteomics, Biochemical/Pharmacological Centre, Philipps-University, 35043, Marburg, Germany

4 Core Facility Bioinformatics, Philipps-University, 35043, Marburg, Germany
5 Department of Cell Biology and Cell Pathology, Philipps-University, 35043, Marburg, Germany

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