PXD046734

PXD046734 is an original dataset announced via ProteomeXchange.

Title	Phosphoprotein dynamics of interacting tumor and T cells by HySic
Description	Functional interactions between cytotoxic T cells and tumor cells are central to anti-cancer immunity. Some of the proteins involved, particularly immune checkpoints expressed by T cells, serve as promising clinical targets in immunotherapy. However, our understanding of the complexity and dynamics of the interactions between tumor cells and T cells is only rudimentary. Here we present HySic (for Hybrid quantification of SILAC (Stable Isotope Labelling by Amino acids in Cell culture)-labeled interacting cells) as an innovative method to quantify protein and phosphorylation dynamics between and within physically interacting (heterotypic) cells. We show that co-cultured HLA/antigen/TCR-matched tumor and T cells engage in stable interactions, allowing for in-depth HySic analysis. This method does not require physical separation of the two cell types for subsequent MS proteome and phosphoproteome measurements using label-free quantification (LFQ). We demonstrate that HySic can be used to identify proteins whose abundance or activation status changes upon interactions between T cells and tumor cells. We validated HySic with established signal transduction pathways, including IFN. Using HySic we also identified the RHO/RAC/PAK1 signaling pathway to be activated upon T cell:tumor cell interaction. Pharmacologic inhibition of PAK1 sensitized tumor cells to T cell killing. Thus, HySic is a simple method to study short-term protein signaling dynamics in physically interacting cells, which can be easily extended to other biological systems.
HostingRepository	PRIDE
AnnounceDate	2023-12-21
AnnouncementXML	Submission_2023-12-20_17:33:51.379.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kelly Stecker
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Q Exactive HF; Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-07 02:55:37	ID requested
⏵ 1	2023-12-20 17:33:51	announced

10.1016/J.CELREP.2023.113598;

submitter keyword: Cell signaling,Phosphoproteomics, Interacting cells, Co-culture, HySic, Human, Cancer, SILAC, Cell Culture, T cells, Intercellular communication

Kelly E.
contact affiliation	Utrecht University, Biomolecular Mass Spectrometry and Proteomics
contact email	k.e.stecker@uu.nl
lab head
Kelly Stecker
contact affiliation	Utrecht University
contact email	k.e.stecker@uu.nl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD046734
     1. Label: PRIDE project
     2. Name: Phosphoprotein dynamics of interacting tumor and T cells by HySic