5 research outputs found
The voltage-gated proton channel HVCN1 modulates mitochondrial ROS production and inflammatory response in macrophages
PhDIt is clear that the voltage-‐gated proton channel HVCN1 plays an essential role in
a range of cell types, in particular immune cells. Previous published work
has confirmed the existence of proton
channels in both murine and human macrophages. However, the role of HVCN1 in
macrophages has not been investigated.
Given that the current literature on
voltage-‐gated proton channels in immune
cells has found HVCN1 to be involved in
several cellular processes (such as the
respiratory burst and signalling events)
it is important to establish its
functional role in macrophages, which are
a crucial constituent of the immune system. The aim of my thesis was to investigate the function of voltage-‐gated proton channels in macrophages with the use of mice with a disrupting mutation within the Hvcn1 gene, which results in HVCN1 loss. In particular, I wanted to address how Hvcn1-‐/-‐ macrophages responded to LPS activation. I hypothesised that HVCN1 regulates the respiratory burst of macrophages and
that it potentially modulates mitochondrial ROS production, and in doing so, may affect several functional aspects of macrophage biology
Dynamic imaging for CAR-T-cell therapy
Abstract Chimaeric antigen receptor (CAR) therapy is entering the mainstream for the treatment of CD19 + cancers. As is does we learn more about resistance to therapy and the role, risks and management of toxicity. In solid tumour CAR therapy research the route to the clinic is less smooth with a wealth of challenges facing translating this, potentially hugely valuable, therapeutic option for patients. As we strive to understand our successes, and navigate the challenges, having a clear understanding of how adoptively transferred CAR-T-cells behave in vivo and in human trials is invaluable. Harnessing reporter gene imaging to enable detection and tracking of small numbers of CAR-T-cells after adoptive transfer is one way by which we can accomplish this. The compatibility of certain reporter gene systems with tracers available routinely in the clinic makes this approach highly useful for future appraisal of CAR-T-cell success in humans. This review covers the research to date in reporter gene imaging of gene-modified T-cells. Chimaeric antigen receptors (CARs) are membranespanning fusion molecules in which a targeting moiety, usually a single chain antibody fragment, is coupled via hinge and transmembrane elements to an activating endodomain. CARs can be transduced into human T-cells, via retroviral or lentiviral vectors, with the potential for co-expression of other genes. When expressed in T-cells CARs redirect their specificity against a designated native 'antigen', obviating the need for either HLA expression or antigen processing. Immunotherapy using CAR-engineered T-cells is acquiring an increasing niche in the experimental therapy of malignant disease [1]. Recent spectacular results have been achieved in the treatment of CD19 + haematologic malignancies + acute lymphoid leukaemia (ALL) Key words: CAR-T-cells, dynamic imaging, PET-CT, reporter genes, SPECT-CT. Abbreviations: CT, computed tomography; CAR, chimaeric antigen receptor; D2R, dopamine type 2-receptor; hdCKDM, human deoxycytidine kinase double mutant; hNET, human noradrenaline transporter gene; hNIS, human sodium iodide symporter; hSSTr2, human somatostatin receptor subtype 2; HSV-tk, herpes simplex virus thymidine kinase; h TK2DM, truncated and mutated human mitochondrial thymidine kinase 2; MIBG, metaiodobenzylguanidine; PET, positron emission tomography; SPECT, single photon emission computed tomography
Clinically Compliant Spatial and Temporal Imaging of Chimeric Antigen Receptor T-cells
Adoptive transfer of chimeric antigen receptor (CAR) T cells has shown promising anticancer results in clinical trials. Here the authors use the human sodium iodide symporter (hNIS) as a reporter gene to image human CAR T cells in cancer-bearing mice using broadly available tracers and imaging platforms
Crosstalk between the canonical NF-κB and Notch signaling pathways inhibits Pparγ expression and promotes pancreatic cancer progression in mice
The majority of human pancreatic cancers have activating mutations in the KRAS proto-oncogene. These mutations result in increased activity of the NF-κB pathway and the subsequent constitutive production of proinflammatory cytokines. Here, we show that inhibitor of κB kinase 2 (Ikk2), a component of the canonical NF-κB signaling pathway, synergizes with basal Notch signaling to upregulate transcription of primary Notch target genes, resulting in suppression of antiinflammatory protein expression and promotion of pancreatic carcinogenesis in mice. We found that in the KrasG12DPdx1-cre mouse model of pancreatic cancer, genetic deletion of Ikk2 in initiated pre-malignant epithelial cells substantially delayed pancreatic oncogenesis and resulted in downregulation of the classical Notch target genes Hes1 and Hey1. Tnf-α stimulated canonical NF-κB signaling and, in collaboration with basal Notch signals, induced optimal expression of Notch targets. Mechanistically, Tnf-α stimulation resulted in phosphorylation of histone H3 at the Hes1 promoter, and this signal was lost with Ikk2 deletion. Hes1 suppresses expression of Pparg, which encodes the antiinflammatory nuclear receptor Pparγ. Thus, crosstalk between Tnf-α/Ikk2 and Notch sustains the intrinsic inflammatory profile of transformed cells. These findings reveal what we believe to be a novel interaction between oncogenic inflammation and a major cell fate pathway and show how these pathways can cooperate to promote cancer progression