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Cook, J. R.

Publications and source records attributed to Cook, J. R..

3 recordsLinked to original sources

Cell volume tunes macrophage innate inflammatory responses through promoting type I interferon signalling.

Macrophages are key effectors in co-ordinating inflammatory and immune responses to threats to the host. How macrophages decipher diverse danger signals to tailor inflammatory responses remains an unanswered question. Cell volume control is critical for normal cellular function. Disturbances in extracellular and intracellular homeostasis induce changes in cell volume, but the impact of disruptions in cell volume in controlling macrophage inflammatory responses is poorly understood. Here, we discover that macrophages use cell volume control as a bona fide danger sensing mechanism to promote and augment inflammatory responses. Using macrophages deficient in the volume regulated anion channel (VRAC), which lack cell volume control under hypo-osmotic conditions, we show that disruptions in cell volume are sensed by macrophages to drive a large transcriptomic response and induction of inflammation. Cell volume disruption, particularly loss of cell volume control, induces type I interferon signalling through a DNA- and STING-dependent mechanism, but independent of cGAS and 23cGAMP transport. Further, we found that cell volume changes synergise with diverse pathogen-mediated signalling to augment type I interferon responses and exacerbate the cytokine storm in a mouse model of hyperinflammation. Our findings highlight cell volume as an important regulator in shaping inflammatory responses, adding to our understanding of how macrophages sense complex danger signals and threats.

immunology↗

Toward a Cell Culture Model of Portal Axis Lipid Handling

The health risks posed by excessive visceral white adipose tissue (WAT) may arise from exposure to deleterious intestinal factors in transit through the portal system. These may include an altered bile acid (BA) pool - particularly with respect to 12-hydroxylated BA (12-HBA), which we have previously demonstrated are closely associated with insulin resistance. However, given the complexity of the enterohepatic milieu, we have worked to develop a co-culture system that allows for controlled study of the interactions between human induced pluripotent stem cell (hiPSC) derived adipocytes and primary human intestinal organoids (hIO). We present preliminary data on the characterization of both cellular compartments and their interactions, particularly in the context of BA treatment. We have recapitulated the basic functionality of each tissue type and have found reciprocal, potentially important changes in gene expression in each. However, we also have encountered numerous technical challenges in the development of this co-culture system and provide methodological observations to assist others seeking to work with a co-culture system of this type.

cell biology↗

CRISPR interference identifies vulnerable cellular pathways with bactericidal phenotypes in Mycobacterium tuberculosis

Mycobacterium tuberculosis remains a leading cause of death for which new drugs are needed. The identification of drug targets has been advanced by high-throughput and targeted genetic deletion strategies. Each though has limitations including the inability to distinguish between levels of vulnerability, lethality and scalability as a molecular tool. Using mycobacterial CRISPR interference in combination with phenotypic screening we have overcome these individual issues to investigate essentiality, vulnerability and lethality for 96 target genes from a diverse array of cellular pathways, many of which are potential antibiotic targets. Essential genes involved in cell wall synthesis and central cellular functions were equally vulnerable and often had bactericidal consequences. Conversely, essential genes involved in metabolism, oxidative phosphorylation or amino acid synthesis were less vulnerable to inhibition and frequently bacteriostatic. In conclusion, this study provides novel insights into mycobacterial genetics and biology that will help to prioritise potential drug targets.

genomics↗