Exploring the Immunomodulatory Potential of Pancreatic Cancer-Derived Extracellular Vesicles Through Proteomic and Functional Analyses

Overview

Journal Cancers (Basel)

Publisher MDPI

Specialty Oncology

Date 2024 May 25

PMID 38791876

Authors

Anna Piro

Maria Concetta Cufaro

Paola Lanuti

Davide Brocco

Laura De Lellis

Rosalba Florio

Serena Pilato

Sara Pagotto

Simone De Fabritiis

Simone Vespa

Giulia Catitti

Fabio Verginelli

Pasquale Simeone

Damiana Pieragostino

Piero Del Boccio

Antonella Fontana

Antonino Grassadonia

Mauro Di Ianni

Alessandro Cama

Serena Veschi

Affiliations

Soon will be listed here.

Abstract

Pancreatic cancer (PC) has a poor prognosis and displays resistance to immunotherapy. A better understanding of tumor-derived extracellular vesicle (EV) effects on immune responses might contribute to improved immunotherapy. EVs derived from Capan-2 and BxPC-3 PC cells isolated by ultracentrifugation were characterized by atomic force microscopy, Western blot (WB), nanoparticle tracking analysis, and label-free proteomics. Fresh PBMCs from healthy donors were treated with PC- or control-derived heterologous EVs, followed by flow cytometry analysis of CD8+ and CD4+ lymphocytes. The proteomics of lymphocytes sorted from EV-treated or untreated PBMCs was performed, and the IFN-γ concentration was measured by ELISA. Notably, most of the proteins identified in Capan-2 and BxPC-3 EVs by the proteomic analysis were connected in a single functional network ( = 1 × 10) and were involved in the "Immune System" (FDR: 1.10 × 10 and 3.69 × 10, respectively). Interestingly, the treatment of healthy donor-derived PBMCs with Capan-2 EVs but not with BxPC-3 EVs or heterologous control EVs induced early activation of CD8+ and CD4+ lymphocytes. The proteomics of lymphocytes sorted from EV-treated PBMCs was consistent with their activation by Capan-2 EVs, indicating IFN-γ among the major upstream regulators, as confirmed by ELISA. The proteomic and functional analyses indicate that PC-EVs have pleiotropic effects, and some may activate early immune responses, which might be relevant for the development of highly needed immunotherapeutic strategies in this immune-cold tumor.

References

Brocco D, De Bellis D, Marino P, Simeone P, Grassadonia A, De Tursi M . High Blood Concentration of Leukocyte-Derived Extracellular Vesicles Is Predictive of Favorable Clinical Outcomes in Patients with Pancreatic Cancer: Results from a Multicenter Prospective Study. Cancers (Basel). 2022; 14(19). PMC: 9562679. DOI: 10.3390/cancers14194748. View

Mulcahy L, Pink R, Carter D . Routes and mechanisms of extracellular vesicle uptake. J Extracell Vesicles. 2014; 3. PMC: 4122821. DOI: 10.3402/jev.v3.24641. View

Ma F, Vayalil J, Lee G, Wang Y, Peng G . Emerging role of tumor-derived extracellular vesicles in T cell suppression and dysfunction in the tumor microenvironment. J Immunother Cancer. 2021; 9(10). PMC: 8513270. DOI: 10.1136/jitc-2021-003217. View

Veschi S, De Lellis L, Florio R, Lanuti P, Massucci A, Tinari N . Effects of repurposed drug candidates nitroxoline and nelfinavir as single agents or in combination with erlotinib in pancreatic cancer cells. J Exp Clin Cancer Res. 2018; 37(1):236. PMC: 6151049. DOI: 10.1186/s13046-018-0904-2. View

Tang D, Chen M, Huang X, Zhang G, Zeng L, Zhang G . SRplot: A free online platform for data visualization and graphing. PLoS One. 2023; 18(11):e0294236. PMC: 10635526. DOI: 10.1371/journal.pone.0294236. View

Han L, Lam E, Sun Y . Extracellular vesicles in the tumor microenvironment: old stories, but new tales. Mol Cancer. 2019; 18(1):59. PMC: 6441234. DOI: 10.1186/s12943-019-0980-8. View

Thery C, Witwer K, Aikawa E, Alcaraz M, Anderson J, Andriantsitohaina R . Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 2019; 7(1):1535750. PMC: 6322352. DOI: 10.1080/20013078.2018.1535750. View

Cossarizza A, Chang H, Radbruch A, Acs A, Adam D, Adam-Klages S . Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition). Eur J Immunol. 2019; 49(10):1457-1973. PMC: 7350392. DOI: 10.1002/eji.201970107. View

Liu C, He D, Li L, Zhang S, Wang L, Fan Z . Extracellular vesicles in pancreatic cancer immune escape: Emerging roles and mechanisms. Pharmacol Res. 2022; 183:106364. DOI: 10.1016/j.phrs.2022.106364. View

10.

Shen T, Huang Z, Shi C, Pu X, Xu X, Wu Z . Pancreatic cancer-derived exosomes induce apoptosis of T lymphocytes through the p38 MAPK-mediated endoplasmic reticulum stress. FASEB J. 2020; 34(6):8442-8458. DOI: 10.1096/fj.201902186R. View

11.

De Lellis L, Florio R, Di Bella M, Brocco D, Guidotti F, Tinari N . Exosomes as Pleiotropic Players in Pancreatic Cancer. Biomedicines. 2021; 9(3). PMC: 8002012. DOI: 10.3390/biomedicines9030275. View

12.

Dassler-Plenker J, Reiners K, van den Boorn J, Hansen H, Putschli B, Barnert S . RIG-I activation induces the release of extracellular vesicles with antitumor activity. Oncoimmunology. 2016; 5(10):e1219827. PMC: 5087302. DOI: 10.1080/2162402X.2016.1219827. View

13.

Buzzai A, Wagner T, Audsley K, Newnes H, Barrett L, Barnes S . Diverse Anti-Tumor Immune Potential Driven by Individual IFNα Subtypes. Front Immunol. 2020; 11:542. PMC: 7145903. DOI: 10.3389/fimmu.2020.00542. View

14.

Legorreta-Haquet M, Santana-Sanchez P, Chavez-Sanchez L, Chavez-Rueda A . The effect of prolactin on immune cell subsets involved in SLE pathogenesis. Front Immunol. 2022; 13:1016427. PMC: 9650038. DOI: 10.3389/fimmu.2022.1016427. View

15.

Marchisio M, Simeone P, Bologna G, Ercolino E, Pierdomenico L, Pieragostino D . Flow Cytometry Analysis of Circulating Extracellular Vesicle Subtypes from Fresh Peripheral Blood Samples. Int J Mol Sci. 2020; 22(1). PMC: 7793062. DOI: 10.3390/ijms22010048. View

16.

Nannan L, Oudart J, Monboisse J, Ramont L, Brassart-Pasco S, Brassart B . Extracellular Vesicle-Dependent Cross-Talk in Cancer-Focus on Pancreatic Cancer. Front Oncol. 2020; 10:1456. PMC: 7466446. DOI: 10.3389/fonc.2020.01456. View

17.

Madonna R, Pieragostino D, Cufaro M, Doria V, Del Boccio P, Deidda M . Ponatinib Induces Vascular Toxicity through the Notch-1 Signaling Pathway. J Clin Med. 2020; 9(3). PMC: 7141219. DOI: 10.3390/jcm9030820. View

18.

Wolfers J, Lozier A, Raposo G, Regnault A, Thery C, Masurier C . Tumor-derived exosomes are a source of shared tumor rejection antigens for CTL cross-priming. Nat Med. 2001; 7(3):297-303. DOI: 10.1038/85438. View

19.

Boukhaled G, Harding S, Brooks D . Opposing Roles of Type I Interferons in Cancer Immunity. Annu Rev Pathol. 2020; 16:167-198. PMC: 8063563. DOI: 10.1146/annurev-pathol-031920-093932. View

20.

Cibrian D, Sanchez-Madrid F . CD69: from activation marker to metabolic gatekeeper. Eur J Immunol. 2017; 47(6):946-953. PMC: 6485631. DOI: 10.1002/eji.201646837. View