- Title Information
- Title
- Phosphoproteomic Analysis of T Cell Receptor and Chimeric Antigen Receptor Signalling: Pathway Discovery and Regulatory Mechanisms
- Type of Resource (primo)
- dissertations
- Name:
Personal
- Name Part
- Callahan, Aurora
- Role
- Role Term:
Text
- creator
- Name:
Personal
- Name Part
- Salomon, Arthur
- Role
- Role Term:
Text
- Advisor
- Name:
Personal
- Name Part
- Fawzi, Nicolas
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- DeLong, Alison
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Beura, Lalit
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- High, Anthony
- Role
- Role Term:
Text
- Reader
- Name:
Corporate
- Name Part
- Brown University. Department of Molecular Biology, Cell Biology and Biochemistry
- Role
- Role Term:
Text
- sponsor
- Origin Information
- Copyright Date
- 2026
- Physical Description
- Extent
- lv, 408 p.
- digitalOrigin
- born digital
- Note:
thesis
- Thesis (Ph. D.)--Brown University, 2026
- Genre (aat)
- theses
- Abstract
- Mass spectrometry (MS) based proteomics has revolutionised biological discovery by allowing for high-throughput, accurate quantification of proteins, posttranslational modifications, and protein-protein interactions. Proteomics has been instrumental in the discovery and characterisation of T cell biology, particularly in phosphorylation-based T cell receptor (TCR) and chimeric antigen receptor (CAR) signalling. Although we understand a great deal about TCR and CAR signalling in T cells, open questions remain regarding the precise regulatory mechanisms involved in activating and deactivating T cells, and whether T cells elicit responses from their target cells. There are three main aims of this work: (1) to evaluate and optimise methodology for identifying tyrosine phosphorylation using MS, (2) to use MS to understand the regulatory networks involved in TCR signalling and activation, and (3) to use MS to determine how CAR design impacts both CAR T cell function and the targeted cells. I determined that suspension-trapping proteins provides optimal recovery of peptides for downstream phosphotyrosine (pY)-MS analysis, and that the BOOST method enhances identification of a subset of low abundance pY sites in primary murine T cells. I showed that three historically redundant protein tyrosine phosphatases uniquely impact global pY abundance and signalling regulation, while two critical protein tyrosine kinases uniquely regulate mRNA regulatory proteins and differentially impact T cell activation. Finally I determined that pY signalling from CARs is largely driven by inclusion of TCRζ, and a native ligand-based CAR can activate pY signalling and induce proliferation in target cells. Together, this work defines workflows for pY site identification using MS, uncovers novel regulation governing T cell pY homeostasis, and identifies potential shortcomings of CAR T cell therapies.
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/00968006")
- Topic
- Immunology
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01011435")
- Topic
- Mass spectrometry
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/00845386")
- Topic
- Cancer--Immunotherapy
- Language
- Language Term (ISO639-2B)
- English
- Record Information
- Record Content Source (marcorg)
- RPB
- Record Creation Date
(encoding="iso8601")
- 20260516