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Demetriou, P.

Publications and source records attributed to Demetriou, P..

5 recordsLinked to original sources

Endothelin receptor blockade potentiates adoptive T cell and CAR T cell therapies

Adoptive T cell therapies have transformed the treatment of selected haematological malignancies but remain limited in many cancer settings by biological barriers that restrict effective and durable antitumour responses. Here, we investigated whether pharmacological blockade of the endothelin receptor pathway, which regulates vascular, stromal and immune processes relevant to antitumour responses, could enhance T cell-based cellular immunotherapy. Using orthotopic 4T1 triple-negative breast cancer and A20 B cell lymphoma models, we evaluated endothelin receptor blockade with non-engineered adoptive T cell therapy and anti-CD19 CAR T cell therapy, respectively, with or without immune checkpoint inhibition. Endothelin receptor blockade substantially increased objective responses across both therapeutic platforms, while the addition of immune checkpoint inhibition further increased response frequency and promoted maintained complete responses, resulting in the most durable tumour control. Mice achieving maintained complete responses resisted tumour rechallenge, consistent with long-term antitumour immune protection. High-dimensional T cell profiling identified distinct intratumoral T cell states associated with tumour control and elevated CD2 expression as a recurring feature across these populations. Together, these findings identify endothelin receptor blockade as a rational combinatorial strategy for enhancing T cell-based cellular immunotherapy and provide a strong rationale for the clinical evaluation of this therapeutic strategy.

immunology↗

CD2 expression is co-regulated with stemness- and exhaustion-associated factors in human T cells

The CD2-CD58 pathway has been highlighted as a major player in anti-tumour T cell immunity. Our study reveals that CD2 costimulation strength significantly correlates with T cell activation, the average number of cell divisions, fold expansion, and IFN-{gamma} production. Our findings suggest that the correlation of CD2 strength with the level of CD25 expression is a potential regulatory mechanism by which CD2 strength enhances above proliferation parameters. We find that human brain cancer tumour-infiltrating CD8+ and CD4+ T cells exhibit reduced levels of CD2, suggestive of a compromised CD2 strength upon CD2 engagement. Through a genome-wide CRISPR-Cas9 knockout screen, we identified two epigenetic regulators, SUZ12 and BAP1, as positive modulators of CD2 expression. We demonstrate that BAP1 is crucial for the upregulation and sustained high expression of CD2 following T cell activation. We reveal that CD2 is co-regulated with other co-stimulatory/inhibitory receptors, and factors associated with T cell stemness and exhaustion, in a dose-dependent manner. Importantly, we rescue the loss of CD2 due to BAP1 knockout by pharmacological inhibition of histone deacetylases making this a harnessable regulatory pathway. The insight from our study enhance our understanding of CD2-mediated T cell regulation and identify essential regulators of this pathway.

immunology↗

In silico characterization of mechanisms positioning costimulatory and checkpoint complexes in immune synapses

Integrin and small immunoglobulin superfamily (sIGSF) adhesion complexes function physiologically in human immunological synapses (IS) wherein sIGSF complexes form a corolla of microdomains around an integrin ring and secretory core. The corolla recruits and retains the major costimulatory and checkpoint complexes that regulate the response to T cell receptor (TCR) engagement, making forces that govern corolla formation of particular interest. We developed a phenomenological agent-based model in order to test different hypotheses concerning the mechanisms underlying molecular reorganization during IS formation. The model showed that sIGSF complexes are passively excluded to the distal aspect of the IS as long as their interaction with the ramified F-actin transport network is absent or weaker than that of integrins. An attractive force between sIGSF adhesion and costimulatory/checkpoint complexes relocates the latter from the centre of the IS to the corolla. The simulations suggest that size based sorting interactions with large glycocalyx components as well as a short-range self-attraction between sIGSF complexes explain the corolla "petals". These molecular and mechanistic features establish a general model that can recapitulate complex pattern formation processes observed in cell-bilayer and cell-cell interfaces. One Sentence SummaryComputer simulations of immunological synapses reveal the localization mechanisms of immunoglobulin superfamily adhesion and costimulatory/checkpoint complexes.

immunology↗

CD2 expression acts as a quantitative checkpoint for immunological synapse structure and T-cell activation.

The CD2 receptor has been described as an adhesion and costimulatory receptor on T cells. Here, transcriptional profiling of colorectal cancers (CRC) revealed a negative correlation between CD2 expression and \"exhausted CD8+ T-cells\" gene signatures. Furthermore, we detected reduced surface CD2 levels in exhausted CD127lowPD-1hi CD3+CD8+ tumour infiltrating lymphocytes (TILs) in CRC. We describe a CD2 expression-level-dependent switch in CD2-CD58 localization between central and peripheral domains in the immunological synapse (IS). A peripheral \"CD2 corolla\" formed when CD2 surface expression was sufficiently high and its cytoplasmic domain intact. The corolla recruited other ligated receptors like CD28, boosted recruitment of activated Src-family kinases (pSrc), LAT and PLC-{gamma} in the IS and consequently T-cell activation in response to a tumour antigen. Corolla formation and pSrc in the IS increased linearly with CD2 expression, whereas pSrc signals were reduced by high, \"exhausted-like\" levels of PD-1, which invaded the corolla. These results suggest two levels of inhibition of Src-family kinases in CD3+CD8+ TILs: reduced CD2 expression and high PD-1 expression.

immunology↗

A tissue-like platform for studying engineered quiescent human T-cells’ interactions with dendritic cells

Research in the field of human immunology is restricted by the lack of a system that reconstitutes the in-situ activation dynamics of quiescent human antigen-specific T-cells interacting with dendritic cells. Here we report a tissue-like system that recapitulates the dynamics of engineered primary human immune cell. Our approach facilitates real-time single cell manipulations, tracking of interactions and functional responses complemented by population-based measurements of cytokines, activation status and proliferation. As a proof of concept, we recapitulate immunological phenomenon such as CD4 help to CD8 T-cells through enhanced maturation of DCs and effect of PD-1 checkpoint blockades. In addition, we characterise unique dynamics of T-cell/DC interactions as a function of antigen affinity.

immunology↗