ADAPT identifies an ESCRT complex composition that discriminates VCaP from LNCaP prostate cancer cell exosomes
Autor: | Zenyu Zhong, Janet E. Duncan, Stephen C. Logie, Xixi Wei, Jelena Zarkovic, Michael Famulok, Varun Maher, Heather A. O'Neill, Günter Mayer, David Spetzler, Teresa T. Tinder, Aniket S. Bondre, Matthew Rosenow, Tassilo Hornung, Mark R. Miglarese, Kimberly M. Fowler, Melissa N. Richards |
---|---|
Rok vydání: | 2020 |
Předmět: |
Male
AcademicSubjects/SCI00010 Biology Exosomes 03 medical and health sciences Prostate cancer 0302 clinical medicine Chemical Biology and Nucleic Acid Chemistry Cell Line Tumor LNCaP Genetics medicine Humans 030304 developmental biology 0303 health sciences Endosomal Sorting Complexes Required for Transport SELEX Aptamer Technique Prostatic Neoplasms Aptamers Nucleotide medicine.disease Phenotype Microvesicles Cell biology ESCRT complex 030220 oncology & carcinogenesis Cancer cell Y-Box-Binding Protein 1 Target protein Systematic evolution of ligands by exponential enrichment |
Zdroj: | Nucleic Acids Research (London) Nucleic Acids Research |
ISSN: | 1362-4962 0305-1048 |
Popis: | Libraries of single-stranded oligodeoxynucleotides (ssODNs) can be enriched for sequences that specifically bind molecules on naïve complex biological samples like cells or tissues. Depending on the enrichment strategy, the ssODNs can identify molecules specifically associated with a defined biological condition, for example a pathological phenotype, and thus are potentially useful for biomarker discovery. We performed ADAPT, a variant of SELEX, on exosomes secreted by VCaP prostate cancer cells. A library of ∼1011 ssODNs was enriched for those that bind to VCaP exosomes and discriminate them from exosomes derived from LNCaP prostate cancer cells. Next-generation sequencing (NGS) identified the best discriminating ssODNs, nine of which were resynthesized and their discriminatory ability confirmed by qPCR. Affinity purification with one of the sequences (Sequence 7) combined with LC–MS/MS identified its molecular target complex, whereof most proteins are part of or associated with the multiprotein ESCRT complex participating in exosome biogenesis. Within this complex, YBX1 was identified as the directly-bound target protein. ADAPT thus is able to differentiate exosomes from cancer cell subtypes from the same lineage. The composition of ESCRT complexes in exosomes from VCaP versus LNCaP cells might constitute a discriminatory element between these prostate cancer subtypes. |
Databáze: | OpenAIRE |
Externí odkaz: |