A5020671002- Ubiquitin and proteasome pathways
- Advanced Proteomics Techniques and Applications
- Immune cells in cancer
- Cellular transport and secretion
- Acute Myeloid Leukemia Research
- Histone Deacetylase Inhibitors Research
- Mass Spectrometry Techniques and Applications
- Myeloproliferative Neoplasms: Diagnosis and Treatment
- Lipid Membrane Structure and Behavior
- Epigenetics and DNA Methylation
- Phagocytosis and Immune Regulation
- Advanced biosensing and bioanalysis techniques
- Cancer, Lipids, and Metabolism
- Cancer, Hypoxia, and Metabolism
- Immunotherapy and Immune Responses
- S100 Proteins and Annexins
- Nuclear Receptors and Signaling
- Macrophage Migration Inhibitory Factor
- RNA modifications and cancer
- Platelet Disorders and Treatments
- Galectins and Cancer Biology
- Reproductive System and Pregnancy
- T-cell and B-cell Immunology
- Acute Lymphoblastic Leukemia research
- Cancer-related Molecular Pathways
Newcastle University
2019-2024
Newcastle upon Tyne Hospital
2024
Memorial Sloan Kettering Cancer Center
2024
Hospital Universitario Fundación Jiménez Díaz
2016-2019
Universidad Autónoma de Madrid
2016-2019
Centro de Biología Molecular Severo Ochoa
2016-2019
Consejo Superior de Investigaciones Científicas
2016
Thermal proteome profiling (TPP) provides a powerful approach to studying proteome-wide interactions of small therapeutic molecules and their target off-target proteins, complementing phenotypic-based drug screens. Detecting differences in thermal stability due engagement requires high quantitative accuracy consistent detection. Isobaric tandem mass tags (TMTs) are used multiplex samples increase quantification precision TPP analysis by data-dependent acquisition (DDA). However, advances...
BMDMs are a key model system to study macrophage biology in vitro. Commonly used methods differentiate macrophages from BM treatment with either recombinant M-CSF or the supernatant of L929 cells, which secrete M-CSF. However, little is known about composition cell-conditioned media (LCCM) and how it affects BMDM phenotype. Here, we quantitative mass spectrometry characterise kinetics protein secretion cells over 2-wk period, identifying 2,193 proteins. Whereas very abundant LCCM, identified...
Abstract Phagocytosis is a key process in innate immunity and homeostasis. After particle uptake, newly formed phagosomes mature by acquisition of endolysosomal enzymes. Macrophage activation interferon gamma (IFN‐γ) increases microbicidal activity, but delays phagosomal maturation an unknown mechanism. Using quantitative proteomics, we show that proteins harbour high levels typical atypical ubiquitin chain types. Moreover, ubiquitylation vesicle trafficking substantially enhanced upon IFN‐γ...
Trilaciclib, a CDK4/6 inhibitor, was approved as myeloprotective agent for protecting bone marrow from chemotherapy-induced damage in extensive-stage small cell lung cancer (ES-SCLC). This is achieved through the induction of temporary halt cycle cells. While it has been studied various types, its potential haematological cancers remains unexplored. research aimed to investigate efficacy trilaciclib cancers. Utilizing mass spectrometry-based proteomics, we examined alterations induced by...
Macrophages engulf pathogens into dynamic phagosomes, which many bacteria manipulate for survival. However, isolating pure pathogen-containing phagosomes remains challenging. Here, we developed a novel flow cytometry-based isolation and ultrasensitive proteomics approach to analyse phagosomal bacterial proteomes from macrophages infected with wild-type (WT) Salmonella enterica serovar Typhimurium (STM), ΔphoP mutant, or dead WT at 30 min 4 hrs post-infection. Our provides higher throughput,...
Summary Despite the Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway being frequently altered in T‐ALL/LBL, no specific therapy has been approved for T‐ALL/LBL patients with constitutive signalling by JAK/STAT, so there is an urgent need to identify members that may be potential therapeutic targets. In present study, we searched JAK/STAT potentially modulated through aberrant methylation identified SOCS3 hypermethylation as a recurrent event T‐ALL/LBL....
T-cell lymphoblastic lymphoma is a haematological disease with an urgent need for reliable prognostic biomarkers that allow therapeutic stratification and dose adjustment. The scarcity of human samples responsible the delayed progress in study clinical management this disease, especially compared acute leukaemia, its leukemic counterpart. In present work, we have determined by immunohistochemistry S194-P-FADD protein significantly reduced cohort 22 from lymphoma. Notably, extent such...
Ubiquitylation is an elaborate post-translational modification involved in all biological processes. Its pleotropic effect driven by the ability to form complex polyubiquitin chain architectures that can influence functions. In this study, we optimised sample preparation and chromatographic separation of Ubiquitin peptides for Absolute Quantification Parallel Reaction Monitoring (Ub-AQUA-PRM). Using refined Ub-AQUA-PRM assay, were able quantify ubiquitin types 10-min LC-MS/MS runs. We used...
Inflammatory responses are important in cancer, particularly the context of monocyte-rich aggressive myeloid neoplasm. We developed a label-free cellular phenotypic drug discovery assay to identify anti-inflammatory drugs human monocytes derived from acute leukemia (AML), by tracking several features ionizing only 2500 cells using matrix-assisted laser desorption/ionization-time flight (MALDI-TOF) mass spectrometry. A proof-of-concept screen showed that BCR-ABL inhibitor nilotinib, but not...
ABSTRACT Trilaciclib, a CDK4/6 inhibitor, was approved as myeloprotective agent for protecting bone marrow from chemotherapy-induced damage in extensive-stage small cell lung cancer (ES-SCLC). This is achieved through the induction of temporary halt cycle cells. While it has been studied various types, its potential haematological cancers remains unexplored. research aimed to investigate efficacy trilaciclib cancers. Utilizing mass spectrometry-based proteomics, we examined alterations...
Abstract Immortalised cell lines analogous to their primary counterparts are fundamental research, particularly when large numbers required. Here we report that immortalisation of bone marrow-derived macrophages using the J2-virus resulted in loss a protein interest, MSR1, wild-type cells by an unknown mechanism. This led us perform in-depth mass spectrometry-based proteomic characterisation common murine macrophage (J774A.1, RAW264.7, and BMA3.1A7), with comparison immortalised (iBMDMs), as...
Inborn errors of T cell development present a pediatric emergency in which timely curative therapy is informed by molecular diagnosis. In 11 affected patients across four consanguineous kindreds, we detected homozygosity for single deleterious missense variant the gene NudC domain–containing 3 ( NUDCD3 ) . Two infants had severe combined immunodeficiency with complete absence and B cells (T - SCID), whereas nine showed classical features Omenn syndrome (OS). Restricted antigen receptor usage...
Immortalised cell lines that mimic their primary counterparts are fundamental to research, particularly when large numbers required. Here, we report immortalisation of bone marrow-derived macrophages (iBMDMs) using the J2 virus resulted in loss a protein interest, MSR1, WT cells by an unknown mechanism. This led us perform in-depth mass spectrometry-based proteomic characterisation common murine macrophage (J774A.1, RAW264.7, and BMA3.1A7), comparison with iBMDMs, as well BMDMs from both...
In the present work, we show that T-cell lymphoblastic lymphoma cells exhibit a reduction of FADD availability in cytoplasm, which may contribute to impaired apoptosis. addition, observe phosphorylation inversely correlates with proliferation capacity and tumor aggressiveness. The resultant balance between FADD-dependent apoptotic non-apoptotic abilities define outcome tumor. Thus, propose expression can be reliable biomarkers prognostic value for T-LBL stratification.
Precursor T-cell lymphoblastic neoplasms are aggressive malignancies in need for more effective and specific therapeutic treatments. A significant fraction of these harbor deletions on the locus 9p21, targeting tumor suppressor CDKN2A but also deleting aconitase 1 (ACO1) gene, a neighboring housekeeping gene involved cytoplasm mitochondrial metabolism. Here we show that reducing activity with fluorocitrate decreases viability neoplasia cells correlation to differential expression. The...
Abstract Thermal proteome profiling (TPP) provides a powerful approach to studying proteome-wide interactions of small therapeutic molecules and their target off-target proteins, complementing phenotypic-based drug screens. Detecting differences in thermal stability due engagement requires high quantitative accuracy consistent detection. Isobaric tandem mass tags (TMT) are used multiplex samples increase quantification precision TPP analysis by data-dependent acquisition (DDA). However,...
Abstract Bone marrow-derived macrophages (BMDMs) are a key model system to study macrophage biology in vitro . Commonly used methods differentiate from bone marrow treatment with either recombinant M-CSF or the supernatant of L929 cells, which secrete M-CSF. However, little is known about composition cell conditioned media (LCCM) and how it affects BMDM phenotype. Here, we quantitative mass spectrometry characterise kinetics protein secretion cells over two-week period, identifying 2,193...
Phagosome acidification and proteolysis are essential processes in the immune response to contain eliminate pathogens. In recent years, there has been an increased desire for a rapid accurate method of assessing these real-time. Here, we outline development multiplexed assay that allows simultaneous monitoring phagosome same sample using silica beads conjugated pHrodo DQ BSA. We describe detail how prepare bi-functional particles show proof concept differentially activated macrophages. This...
ABSTRACT Inflammatory responses are important in cancer, particularly the context of monocyte-rich aggressive myeloid neoplasm. We developed a label-free cellular phenotypic drug discovery assay to identify anti-inflammatory drugs human monocytes derived from acute leukaemia (AML), by tracking several biological features ionizing only 2,500 cells using matrix-assisted laser desorption/ionization-time flight (MALDI-TOF) mass spectrometry. A proof-of-concept screen showed that BCR-ABL...
Summary Phagocytosis is a key process in innate immunity and homeostasis. After uptake, newly formed phagosomes mature by acquisition of endo-lysosomal enzymes. Macrophage activation interferon-gamma (IFN-γ) increases microbicidal activity, but delays phagosomal maturation an unknown mechanism. Using quantitative proteomics, we show that proteins harbour high levels typical atypical ubiquitin chain types. Moreover, ubiquitylation vesicle trafficking substantially enhanced upon IFN-γ...