Plasma-Derived Extracellular Vesicles Convey Protein Signatures that Reflect Pathophysiology in Lung and Pancreatic Adenocarcinomas.

Johannes F Fahrmann,Michela Capello,Hiroyuki Katayama,Jody Vykoukal,Ehsan Irajizad,Samir M Hanash,Ichidai Tanaka,Taketo Kato,Xiangying Mao,Anirban Maitra,Ignacio I Wistuba,Edwin J Ostrin

doi:10.3390/cancers12051147

Johannes F Fahrmann, Michela Capello + Show 10 more

Open Access

https://doi.org/10.3390/cancers12051147

Copy DOI

Abstract

Using a combination of mass-spectrometry and aptamer array-based proteomics, we characterized the protein features of circulating extracellular vesicles (EVs) in the context of lung (LUAD) and pancreatic ductal (PDAC) adenocarcinomas. We profiled EVs isolated from conditioned media of LUAD and PDAC cell lines to identify EV-associated protein cargoes released by these cancer cell types. Analysis of the resulting data identified LUAD and PDAC specific and pan-adenocarcinoma EV protein signatures. Bioinformatic analyses confirmed enrichment of proteins annotated to vesicle-associated processes and intracellular compartments, as well as representation of cancer hallmark functions and processes. Analysis of upstream regulator networks indicated significant enrichment of TP53, MYC, TGFB1 and KRAS-driven network effectors (p = 1.69 × 10−77–2.93 × 10−49) manifest in the adenocarcinoma sEV protein cargoes. We extended these findings by profiling the proteome of EVs isolated from lung (N = 15) and pancreatic ductal (N = 6) adenocarcinoma patient plasmas obtained at time of diagnosis, along with EVs derived from matched healthy controls (N = 21). Exploration of these proteomic data revealed abundant protein features in the plasma EVs with capacity to distinguish LUAD and PDAC cases from controls, including features yielding higher performance in the plasma EV isolates relative to unfractionated plasmas.

Highlights

Extracellular vesicles (EVs) encompass a diverse group of lipid-bound nanoparticles, 50–1000 nm in diameter that are released by most cell types in normal as well as diseased states [1,2]
We explored for protein signatures of lung and pancreatic adenocarcinoma that are communicated by extracellular vesicles
We first established profiles of small extracellular vesicles (sEVs)-associated proteins originating from cancer cells through proteomic characterization of sEVs derived from conditioned-media of lung (LUAD) and pancreatic ductal (PDAC) adenocarcinoma cell lines. sEVs were isolated by differential ultracentrifugation [24] of concentrated conditioned medias derived from a panel of ten cell lines (NCI-H23, NCI-H647, NCI-H1573, HCC4019, CFPAC-1, HPAF-II, SU.86.86, Panc 03.27, MIA PaCa-2 and PANC-1)

Summary

Introduction

Extracellular vesicles (EVs) encompass a diverse group of lipid-bound nanoparticles, 50–1000 nm in diameter that are released by most cell types in normal as well as diseased states [1,2]. Cells release non-vesicle lipid structures including lipoproteins and other lipid-protein complexes and varied complexes of proteins, nucleic acids and other biomolecules. These vesicle and non-vesicle entities are actively being categorized and investigated and are currently of considerable research interest [4,5]. We previously demonstrated that EVs disseminated from pancreatic cancer cells communicate a considerable repertoire of tumor antigens that are associated with host humoral immune response and circulating anti-tumor autoantibodies in pancreatic cancer patient plasmas [10]

Methods

Results

Discussion

Conclusion