Multiplexed Relative Quantitation with Isobaric Tagging Mass Spectrometry Reveals Class I Major Histocompatibility Complex Ligand Dynamics in Response to Doxorubicin.
J Patrick MurphyQijia YuPrathyusha KondaJoao A PauloMark P JedrychowskiDaniel J KowalewskiHeiko SchusterYoura KimDerek ClementsAditya JainStefan StevanovicSteven P GygiJoseph D ManciasShashi GujarPublished in: Analytical chemistry (2019)
MHC-I peptides are intracellular-cleaved peptides, usually 8-11 amino acids in length, which are presented on the cell surface and facilitate CD8+ T cell responses. Despite the appreciation of CD8+ T-cell antitumor immune responses toward improvement in patient outcomes, the MHC-I peptide ligands that facilitate the response are poorly described. Along these same lines, although many therapies have been recognized for their ability to reinvigorate antitumor CD8+ T-cell responses, whether these therapies alter the MHC-I peptide repertoire has not been fully assessed due to the lack of quantitative strategies. We develop a multiplexing platform for screening therapy-induced MHC-I ligands by employing tandem mass tags (TMTs). We applied this approach to measuring responses to doxorubicin, which is known to promote antitumor CD8+ T-cell responses during its therapeutic administration in cancer patients. Using both in vitro and in vivo systems, we show successful relative quantitation of MHC-I ligands using TMT-based multiplexing and demonstrate that doxorubicin induces MHC-I peptide ligands that are largely derived from mitotic progression and cell-cycle proteins. This high-throughput MHC-I ligand discovery approach may enable further explorations to understand how small molecules and other therapies alter MHC-I ligand presentation that may be harnessed for CD8+ T-cell-based immunotherapies.
Keyphrases
- cell cycle
- mass spectrometry
- high throughput
- immune response
- drug delivery
- amino acid
- cell proliferation
- liquid chromatography
- ms ms
- cell surface
- liquid chromatography tandem mass spectrometry
- high performance liquid chromatography
- inflammatory response
- mesenchymal stem cells
- drug induced
- endothelial cells
- reactive oxygen species
- simultaneous determination
- solid phase extraction
- toll like receptor
- cell therapy
- capillary electrophoresis
- stress induced