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Proteomic response and variability of surface Vlh-antigens of bacteria Mycoplasma gallisepticum in the eukaryotic invasion model

ABSTRACT: Proteomic response and variability of surface Vlh-antigens of bacteria Mycoplasma gallisepticum in the eukaryotic invasion model

INSTRUMENT(S): QQQ LC/MS 8030 Shimadzu

ORGANISM(S): Mycoplasma Gallisepticum S6 (ncbitaxon:1006581)

SUBMITTER: Dariya Matushkina

PROVIDER: MSV000079642 | MassIVE | Fri Apr 08 15:13:00 BST 2016

REPOSITORIES: MassIVE

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Project description:Background: Despite advancements in cancer immunotherapy, solid tumors remain formidable challenges. In glioma, profound inter- and intra-tumoral heterogeneity of antigen landscape hampers therapeutic development. Therefore, it is critical to consider alternative sources to expand the repertoire of targetable (neo-)antigens and improve therapeutic outcomes. Accumulating evidence suggests that tumor-specific alternative splicing (AS) could be an untapped reservoir of antigens. Results: In this study, we investigated tumor-specific AS events in glioma, focusing on those predicted to generate major histocompatibility complex (MHC)-presentation-independent, cell-surface antigens that could be targeted by antibodies and chimeric antigen receptor (CAR)-T cells. We systematically analyzed bulk RNA-sequencing datasets comparing 429 tumor samples (from The Cancer Genome Atlas [TCGA]) and 9,166 normal tissue samples (from the Genotype-Tissue Expression project [GTEx]), and identified 13 AS events in 7 genes predicted to be expressed in more than 10% of the patients, including PTPRZ1 and BCAN, which were corroborated by an external RNA-sequencing dataset. Subsequently, we validated our predictions and elucidated the complexity of the isoforms using full-length transcript amplicon sequencing on patient-derived glioblastoma cells. However, analyses of the RNA-sequencing datasets of spatially mapped and longitudinally collected clinical tumor samples unveiled remarkable spatiotemporal heterogeneity of the candidate AS events. Furthermore, proteomics analysis did not reveal any peptide spectra matching the putative antigens. Conclusions: Our investigation illustrated the diverse characteristics of the tumor-specific AS events and the challenges of antigen exploration due to their notable spatiotemporal heterogeneity and elusive nature at the protein levels. Redirecting future efforts toward intracellular, MHC-presented antigens could offer a more viable avenue.

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Proteomic response and variability of surface Vlh-antigens of bacteria Mycoplasma gallisepticum in the eukaryotic invasion model

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