{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T23:25:37Z","timestamp":1792106737403,"version":"4.4.0"},"reference-count":40,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2024,12,21]],"date-time":"2024-12-21T00:00:00Z","timestamp":1734739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Postdoctoral Fellowship","award":["PF-23-1153194-01-CDP"],"award-info":[{"award-number":["PF-23-1153194-01-CDP"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Introduction: Prostate cancer treatment has been revolutionized by targeted therapies, including PARP inhibitors, checkpoint immunotherapies, and PSMA-targeted radiotherapies. Despite such advancements, accurate patient stratification remains a challenge, with current methods relying on genomic markers, tissue staining, and imaging. Extracellular vesicle (EV)-derived proteins offer a novel non-invasive alternative for biomarker discovery, holding promise for improving treatment precision. However, the characterization of plasma-derived EVs in prostate cancer patients remains largely unexplored. Methods: We conducted proteomic analyses on EVs isolated from plasma in 27 metastatic castration-resistant prostate cancer (mCRPC) patients. EVs were purified using ultracentrifugation and analyzed via mass spectrometry. Proteomic data were correlated with clinical markers such as serum prostate-specific antigen (PSA) and bone lesion counts. Statistical significance was assessed using Mann\u2013Whitney t-tests and Spearman correlation. Results: The median age of patients was 74 (range: 44\u201394) years. At the time of blood collection, the median PSA level was 70 (range: 0.5\u20131000) ng\/mL. All patients had bone metastasis. A total of 5213 proteins were detected, including EV-related proteins (CD9, CD81, CD63, FLOT1, TSG101) and cancer-related proteins (PSMA, B7-H3, PD-L1). Proteomic profiling of plasma EVs revealed a significant correlation between specific EV-derived proteins and clinical prognostic markers. B7-H3, LAT1, and SLC29A1 showed a strong association with serum PSA levels and number of bone lesions, indicating potential for these proteins to serve as biomarkers of disease burden and therapy response. Conclusions: Our findings demonstrate the potential of EV-based proteomics for identifying biomarkers in mCRPC patients. Proteins such as B7-H3 and LAT1 could guide precision oncology approaches, improving patient stratification. Future research incorporating outcomes data and EV subpopulation analysis is needed to establish clinical relevance.<\/jats:p>","DOI":"10.3390\/cancers16244261","type":"journal-article","created":{"date-parts":[[2024,12,23]],"date-time":"2024-12-23T09:13:38Z","timestamp":1734945218000},"page":"4261","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Isolation of Plasma Extracellular Vesicles for High-Depth Analysis of Proteomic Biomarkers in Metastatic Castration-Resistant Prostate Cancer Patients"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0460-3039","authenticated-orcid":false,"given":"Ali T.","family":"Arafa","sequence":"first","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3133-3679","authenticated-orcid":false,"given":"Megan","family":"Ludwig","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3345-9208","authenticated-orcid":false,"given":"Onur","family":"Tuncer","sequence":"additional","affiliation":[{"name":"Nuclear Medicine Division, Department of Radiology, University of Minnesota Medical School, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lily","family":"Kollitz","sequence":"additional","affiliation":[{"name":"Nuclear Medicine Division, Department of Radiology, University of Minnesota Medical School, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ava","family":"Gustafson","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ella","family":"Boytim","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4504-0033","authenticated-orcid":false,"given":"Christine","family":"Luo","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Sabal","sequence":"additional","affiliation":[{"name":"Nuclear Medicine Division, Department of Radiology, University of Minnesota Medical School, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7410-3630","authenticated-orcid":false,"given":"Daniel","family":"Steinberger","sequence":"additional","affiliation":[{"name":"Nuclear Medicine Division, Department of Radiology, University of Minnesota Medical School, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingchun","family":"Zhao","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott M.","family":"Dehm","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Urology, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuzan","family":"Cayci","sequence":"additional","affiliation":[{"name":"Nuclear Medicine Division, Department of Radiology, University of Minnesota Medical School, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1686-7103","authenticated-orcid":false,"given":"Justin","family":"Hwang","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Division of Hematology\/Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0067-3083","authenticated-orcid":false,"given":"Peter W.","family":"Villalta","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuel S.","family":"Antonarakis","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Division of Hematology\/Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin M.","family":"Drake","sequence":"additional","affiliation":[{"name":"Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA"},{"name":"Department of Urology, University of Minnesota, Minneapolis, MN 55455, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2091","DOI":"10.1056\/NEJMoa1911440","article-title":"Olaparib for metastatic castration-resistant prostate cancer","volume":"382","author":"Mateo","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1200\/JCO.20.01035","article-title":"Rucaparib in men with metastatic castration-resistant prostate cancer harboring a BRCA1 or BRCA2 gene alteration","volume":"38","author":"Abida","year":"2020","journal-title":"J. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1056\/NEJMoa2107322","article-title":"Lutetium-177\u2013PSMA-617 for metastatic castration-resistant prostate cancer","volume":"385","author":"Sartor","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1748","DOI":"10.1200\/JCO.2012.43.1882","article-title":"Germline BRCA mutations are associated with higher risk of nodal involvement, distant metastasis, and poor survival outcomes in prostate cancer","volume":"31","author":"Castro","year":"2013","journal-title":"J. Clin. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.2967\/jnumed.122.265242","article-title":"PSMA PET tumor-to-salivary gland ratio to predict response to [177Lu]PSMA radioligand therapy: An international multicenter retrospective study","volume":"64","author":"Hotta","year":"2023","journal-title":"J. Nucl. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1001\/jamaoncol.2024.0591","article-title":"Metastatic hormone-sensitive prostate cancer and combination treatment outcomes: A review","volume":"10","author":"Hussain","year":"2024","journal-title":"JAMA Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1056\/NEJMra1701695","article-title":"Metastatic prostate cancer","volume":"378","author":"Sartor","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1016\/j.eururo.2015.06.039","article-title":"Metastatic prostate cancer and the bone: Significance and therapeutic options","volume":"68","author":"Gartrell","year":"2015","journal-title":"Eur. Urol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s12943-022-01509-9","article-title":"Exosomes as a new frontier of cancer liquid biopsy","volume":"21","author":"Yu","year":"2022","journal-title":"Mol. Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1016\/j.annonc.2021.01.074","article-title":"Exosome-based liquid biopsies in cancer: Opportunities and challenges","volume":"32","author":"Yu","year":"2021","journal-title":"Ann. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Tang, H., Yu, D., Zhang, J., Wang, M., Fu, M., Qian, Y., Zhang, X., Ji, R., Gu, J., and Zhang, X. (2024). The new advance of exosome-based liquid biopsy for cancer diagnosis. J. Nanobiotechnol., 22.","DOI":"10.1186\/s12951-024-02863-0"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/978-3-030-67171-6_1","article-title":"Introduction to the community of extracellular vesicles","volume":"97","author":"Fonseka","year":"2021","journal-title":"Subcell. Biochem."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Chen, Y.F., Luh, F., Ho, Y.S., and Yen, Y. (2024). Exosomes: A review of biologic function, diagnostic and targeted therapy applications, and clinical trials. J. Biomed. Sci., 31.","DOI":"10.1186\/s12929-024-01055-0"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1007\/s12032-021-01491-0","article-title":"Exosomes and cancer: From molecular mechanisms to clinical applications","volume":"38","author":"Jafari","year":"2021","journal-title":"Med. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Liu, Y., Liu, H., and Tang, W.H. (2019). Exosomes: Biogenesis, biologic function and clinical potential. Cell Biosci., 9.","DOI":"10.1186\/s13578-019-0282-2"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Helwa, I., Cai, J., Drewry, M.D., Zimmerman, A., Dinkins, M.B., Khaled, M.L., Seremwe, M., Dismuke, W.M., Bieberich, E., and Stamer, W.D. (2017). A comparative study of serum exosome isolation using differential ultracentrifugation and three commercial reagents. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0170628"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Sidhom, K., Obi, P.O., and Saleem, A. (2020). A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option. Int. J. Mol. Sci., 21.","DOI":"10.20944\/preprints202007.0485.v2"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kowkabany, G., and Bao, Y. (2024). Nanoparticle Tracking Analysis: An Effective Tool to Characterize Extracellular Vesicles. Molecules, 29.","DOI":"10.3390\/molecules29194672"},{"key":"ref_19","first-page":"E10988","article-title":"The in silico human surfaceome","volume":"115","author":"Goldmann","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e12404","DOI":"10.1002\/jev2.12404","article-title":"Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches","volume":"13","author":"Welsh","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/978-1-0716-2823-2_5","article-title":"Exosomal MicroRNAs: Comprehensive Methods from Exosome Isolation to miRNA Extraction and Purity Analysis","volume":"2595","author":"Muro","year":"2023","journal-title":"Methods Mol. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1021\/acs.jproteome.3c00614","article-title":"In-Depth Proteome Profiling of Small Extracellular Vesicles Isolated from Cancer Cell Lines and Patient Serum","volume":"23","author":"Abhange","year":"2024","journal-title":"J. Proteome Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1038\/s41596-018-0082-x","article-title":"Single-pot, solid-phase-enhanced sample preparation for proteomics experiments","volume":"14","author":"Hughes","year":"2019","journal-title":"Nat. Protoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"888","DOI":"10.1038\/s41591-023-02284-w","article-title":"Neoadjuvant enoblituzumab in localized prostate cancer: A single-arm, phase 2 trial","volume":"29","author":"Shenderov","year":"2023","journal-title":"Nat. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"746","DOI":"10.1016\/j.annonc.2021.03.005","article-title":"Sacituzumab govitecan, a Trop-2-directed antibody-drug conjugate, for patients with epithelial cancer: Final safety and efficacy results from the phase I\/II IMMU-132-01 basket trial","volume":"32","author":"Bardia","year":"2021","journal-title":"Ann. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1038\/s41698-022-00323-2","article-title":"Integrative molecular analyses define correlates of high B7-H3 expression in metastatic castrate-resistant prostate cancer","volume":"6","author":"Shi","year":"2022","journal-title":"NPJ Precis. Oncol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1478","DOI":"10.1016\/j.ajpath.2018.02.014","article-title":"Comprehensive evaluation of programmed death-ligand 1 expression in primary and metastatic prostate cancer","volume":"188","author":"Haffner","year":"2018","journal-title":"Am. J. Pathol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Xu, M., Evans, L., Bizzaro, C.L., Quaglia, F., Verrillo, C.E., Li, L., Stieglmaier, J., Schiewer, M.J., Languino, L.R., and Kelly, W.K. (2022). STEAP1\u20134 (Six-Transmembrane Epithelial Antigen of the Prostate 1\u20134) and Their Clinical Implications for Prostate Cancer. Cancers, 14.","DOI":"10.20944\/preprints202207.0464.v1"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1038\/s41698-024-00599-6","article-title":"Assessment of TROP2, CEACAM5 and DLL3 in metastatic prostate cancer: Expression landscape and molecular correlates","volume":"8","author":"Ajkunic","year":"2024","journal-title":"NPJ Precis. Oncol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4481","DOI":"10.1158\/0008-5472.CAN-15-3376","article-title":"Genetic disruption of the multifunctional CD98\/LAT1 complex demonstrates the key role of essential amino acid transport in the control of mTORC1 and tumor growth","volume":"76","author":"Cormerais","year":"2016","journal-title":"Cancer Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Zhao, X., Sakamoto, S., Wei, J., Pae, S., Saito, S., Sazuka, T., Imamura, Y., Anzai, N., and Ichikawa, T. (2023). Contribution of the L-Type Amino Acid Transporter Family in the Diagnosis and Treatment of Prostate Cancer. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24076178"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1186\/s13045-022-01364-7","article-title":"Immune checkpoint of B7-H3 in cancer: From immunology to clinical immunotherapy","volume":"15","author":"Zhao","year":"2022","journal-title":"J. Hematol. Oncol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1227","DOI":"10.1158\/1078-0432.CCR-20-2584","article-title":"B7-H3: An Attractive Target for Antibody-based Immunotherapy","volume":"27","author":"Kontos","year":"2021","journal-title":"Clin. Cancer Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"eadf6724","DOI":"10.1126\/scitranslmed.adf6724","article-title":"Immune checkpoint B7-H3 is a therapeutic vulnerability in prostate cancer harboring PTEN and TP53 deficiencies","volume":"15","author":"Shi","year":"2023","journal-title":"Sci. Transl. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.eururo.2022.09.004","article-title":"B7-H3 as a Therapeutic Target in Advanced Prostate Cancer","volume":"83","author":"Guo","year":"2023","journal-title":"Eur. Urol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1369","DOI":"10.1158\/2767-9764.CRC-23-0546","article-title":"Pan-Cancer Interrogation of B7-H3 (CD276) as an Actionable Therapeutic Target Across Human Malignancies","volume":"4","author":"Miller","year":"2024","journal-title":"Cancer Res. Commun."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"5205","DOI":"10.1021\/acs.molpharmaceut.4c00597","article-title":"Development of a Specifically Labeled 89Zr Antibody for the Noninvasive Imaging of Tumors Overexpressing B7-H3","volume":"21","author":"Zheng","year":"2024","journal-title":"Mol. Pharm."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1159\/000509434","article-title":"Clinical Characteristics of Metastatic Prostate Cancer Patients Infected with COVID-19 in South Italy","volume":"98","author":"Buonerba","year":"2020","journal-title":"Oncology"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e004424","DOI":"10.1136\/jitc-2021-004424","article-title":"Dual checkpoint targeting of B7-H3 and PD-1 with enoblituzumab and pembrolizumab in advanced solid tumors: Interim results from a multicenter phase I\/II trial","volume":"10","author":"Aggarwal","year":"2022","journal-title":"J. Immunother. Cancer"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"H\u00e4fliger, P., and Charles, R.P. (2019). The L-Type Amino Acid Transporter LAT1\u2014An Emerging Target in Cancer. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20102428"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/16\/24\/4261\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:57:39Z","timestamp":1760115459000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/16\/24\/4261"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,21]]},"references-count":40,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["cancers16244261"],"URL":"https:\/\/doi.org\/10.3390\/cancers16244261","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,21]]}}}