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BLEB COSMONIOUS HIGH SOFTWARE

CD63 was expressed specifically in nano-sized EVs, and GOT1 in large EVs. (B) Protein lysates from cells, large and nano-sized EVs were blotted with the indicated antibodies.

BLEB COSMONIOUS HIGH SOFTWARE

(A) Functional analysis using FunRich software indicates the biological pathways overrepresented either in large (10,000 g) or nano-sized EVs (100,000 g). SILAC Proteomics amoeboid blebbing cancer metabolism extracellular vesicles tumor progression. These observations indicate that LO represent a discrete EV type that may play a distinct role in tumor progression and that may be a source of cancer-specific markers. Among the proteins enriched in LO, cytokeratin 18 (CK18) was one of the most abundant (within the top 5th percentile) and was used to develop an assay to detect LO in the circulation and tissues of mice and patients with prostate cancer. Fluorescent microscopy of large EVs revealed an appearance consistent with LO morphology, indicating that these structures can be categorized as LO. Large EVs exhibited discrete buoyant densities in iodixanol (OptiPrep(TM)) gradients. Glutamine metabolism was altered in cancer cells exposed to large EVs, an effect that was not observed upon treatment with exosomes. Proteins enriched in large EVs included enzymes involved in glucose, glutamine and amino acid metabolism, all metabolic processes relevant to cancer. We report that 25% of the proteins, identified by a quantitative proteomics analysis, are differentially represented in large and nano-sized EVs from prostate cancer cells. Large oncosomes (LO) are atypically large (1-10 µm diameter) cancer-derived extracellular vesicles (EVs), originating from the shedding of membrane blebs and associated with advanced disease.

9 Department of Biochemistry, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Australia.

8 Department of Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea.

7 Vancouver Prostate Centre, Department of Urologic Sciences, University of British Columbia, BC, Canada.

6 Experimental Pharmacology Unit, Department of Research, IRCCS-Istituto Nazionale Tumori G.

5 Department of Experimental and Clinical Biomedical Science, University of Florence, Florence, Italy.

4 Women's Cancer Program, Cedars-Sinai Medical Center, Los Angeles, CA, USA.

3 Department of Surgery, Harvard Medical School, Boston, MA, USA.

2 The Urological Diseases Research Center, Boston Children's Hospital, Boston, MA, USA.

1 Division of Cancer Biology and Therapeutics, Departments of Surgery, Biomedical Sciences and Pathology and Laboratory Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA.