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Comparative proteomic analysis of three major extracellular classes secreted from human adenocarcinoma and metastatic colorectal cancer cells: exosomes, microparticles and shed midbody remnants
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  • Wittaya Suwakulsiri,
  • Rong Xu,
  • alin rai,
  • Maoshan Chen,
  • Adnan Shafiq,
  • David Greening,
  • Richard Simpson
Wittaya Suwakulsiri
Monash University
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Rong Xu
Monash University The Australian Centre for Blood Diseases
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alin rai
Baker Heart and Diabetes Institute
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Maoshan Chen
Army Medical University
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Adnan Shafiq
La Trobe University
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David Greening
Baker Heart and Diabetes Institute
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Richard Simpson
La Trobe University

Corresponding Author:[email protected]

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Cell-derived extracellular vesicles (EVs) are evolutionary-conserved secretory organelles that, based on their molecular composition, are important intercellular signaling regulators. At least three classes of circulating EVs are known based on mechanism of biogenesis: exosomes (sEVs/Exos), microparticles (lEVs/MPs) and shed midbody remnants (sMB-Rs). sEVs/Exos are of endosomal pathway origin, microparticles (lEVs/MPs) from plasma membrane blebbing, and shed midbody remnants (sMB-Rs) arise from symmetric cytokinetic abscission. Here, we isolate sEVs/Exos, lEVs/MPs and sMB-Rs secreted from human isogenic primary (SW480) and metastatic (SW620) colorectal cancer (CRC) cell lines in milligram quantities for label-free MS/MS-based proteomic profiling. Purified EVs revealed selective composition packaging of exosomal protein markers in SW480/SW620-sEVs/Exos, metabolic enzymes in SW480/SW620-lEVs/MPs, while centralspindlin complex proteins, nucleoproteins, splicing factors, RNA granule proteins, translation-initiation factors, and mitochondrial proteins selectively traffic to SW480/SW620-sMB-Rs. Collectively, we identify 39 human cancer-associated genes in EVs; 17 associated with SW480-EVs, 22 with SW620-EVs. We highlight oncogenic receptors/transporters selectively enriched in sEVs/Exos (EGFR/ FAS in SW480-Exos and MET, TGFBR2, ABCB1 in SW620-sEVs/Exos). Interestingly, MDK, STAT1, and TGM2 are selectively enriched in SW480-sMB-Rs, and ADAM15 to SW620-sMB-Rs. Our study reveals sEVs/Exos, lEVs/MPs and sMB-Rs have distinct protein signatures that open potential diagnostic avenues of distinct types of EVs for clinical utility.
02 May 2023Submitted to PROTEOMICS
03 May 2023Assigned to Editor
03 May 2023Submission Checks Completed
03 May 2023Review(s) Completed, Editorial Evaluation Pending
03 May 2023Reviewer(s) Assigned
30 May 2023Editorial Decision: Revise Minor
03 Jul 2023Review(s) Completed, Editorial Evaluation Pending
03 Jul 20231st Revision Received
04 Jul 2023Editorial Decision: Accept