High-Density, Targeted Monitoring of Tyrosine Phosphorylation Reveals Activated Signaling Networks in Human Tumors.
A549 Cells
CD8-Positive T-Lymphocytes
/ immunology
Chromatography, Liquid
/ methods
Colorectal Neoplasms
/ immunology
ErbB Receptors
/ metabolism
Humans
Mass Spectrometry
/ methods
Phosphopeptides
/ analysis
Phosphorylation
Protein Interaction Maps
Protein Processing, Post-Translational
Proteome
/ analysis
Proteomics
/ methods
Signal Transduction
Tyrosine
/ metabolism
Journal
Cancer research
ISSN: 1538-7445
Titre abrégé: Cancer Res
Pays: United States
ID NLM: 2984705R
Informations de publication
Date de publication:
01 05 2021
01 05 2021
Historique:
received:
13
11
2020
revised:
17
12
2020
accepted:
22
01
2021
pubmed:
30
1
2021
medline:
18
9
2021
entrez:
29
1
2021
Statut:
ppublish
Résumé
Tyrosine phosphorylation (pTyr) plays a pivotal role in signal transduction and is commonly dysregulated in cancer. As a result, profiling tumor pTyr levels may reveal therapeutic insights critical to combating disease. Existing discovery and targeted mass spectrometry-based methods used to monitor pTyr networks involve a tradeoff between broad coverage of the pTyr network, reproducibility in target identification across analyses, and accurate quantification. To address these limitations, we developed a targeted approach, termed "SureQuant pTyr," coupling low input pTyr enrichment with a panel of isotopically labeled internal standard peptides to guide data acquisition of low-abundance tyrosine phosphopeptides. SureQuant pTyr allowed for reliable quantification of several hundred commonly dysregulated pTyr targets with high quantitative accuracy, improving the robustness and usability of targeted mass spectrometry assays. We established the clinical applicability of SureQuant pTyr by profiling pTyr signaling levels in human colorectal tumors using minimal sample input, characterizing patient-specific oncogenic-driving mechanisms. While in some cases pTyr profiles aligned with previously reported proteomic, genomic, and transcriptomic molecular characterizations, we highlighted instances of new insights gained using pTyr characterization and emphasized the complementary nature of pTyr measurements with traditional biomarkers for improving patient stratification and identifying therapeutic targets. The turn-key nature of this approach opens the door to rapid and reproducible pTyr profiling in research and clinical settings alike and enables pTyr-based measurements for applications in precision medicine. SIGNIFICANCE: SureQuant pTyr is a mass spectrometry-based targeted method that enables sensitive and selective targeted quantitation of several hundred low-abundance tyrosine phosphorylated peptides commonly dysregulated in cancer, including oncogenic signaling networks.
Identifiants
pubmed: 33509940
pii: 0008-5472.CAN-20-3804
doi: 10.1158/0008-5472.CAN-20-3804
pmc: PMC8137532
mid: NIHMS1669161
doi:
Substances chimiques
Phosphopeptides
0
Proteome
0
Tyrosine
42HK56048U
EGFR protein, human
EC 2.7.10.1
ErbB Receptors
EC 2.7.10.1
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2495-2509Subventions
Organisme : NIEHS NIH HHS
ID : P42 ES027707
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA014051
Pays : United States
Organisme : NCI NIH HHS
ID : U01 CA238720
Pays : United States
Organisme : NIEHS NIH HHS
ID : T32 ES007020
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM087237
Pays : United States
Organisme : NCI NIH HHS
ID : U54 CA210180
Pays : United States
Informations de copyright
©2021 American Association for Cancer Research.
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