Endothelial sensing of AHR ligands regulates intestinal homeostasis.

Endothelial cells line the blood and lymphatic vasculature, and act as an essential physical barrier, control nutrient transport, facilitate tissue immunosurveillance and coordinate angiogenesis and lymphangiogenesis1,2. In the intestine, dietary and microbial cues are particularly important in the regulation of organ homeostasis. However, whether enteric endothelial cells actively sense and integrate such signals is currently unknown. Here we show that the aryl hydrocarbon receptor (AHR) acts as a critical node for endothelial cell sensing of dietary metabolites in adult mice and human primary endothelial cells. We first established a comprehensive single-cell endothelial atlas of the mouse small intestine, uncovering the cellular complexity and functional heterogeneity of blood and lymphatic endothelial cells. Analyses of AHR-mediated responses at single-cell resolution identified tissue-protective transcriptional signatures and regulatory networks promoting cellular quiescence and vascular normalcy at steady state. Endothelial AHR deficiency in adult mice resulted in dysregulated inflammatory responses and the initiation of proliferative pathways. Furthermore, endothelial sensing of dietary AHR ligands was required for optimal protection against enteric infection. In human endothelial cells, AHR signalling promoted quiescence and restrained activation by inflammatory mediators. Together, our data provide a comprehensive dissection of the effect of environmental sensing across the spectrum of enteric endothelia, demonstrating that endothelial AHR signalling integrates dietary cues to maintain tissue homeostasis by promoting endothelial cell quiescence and vascular normalcy.

Validation of RAH VQ SPECT/CT lobar quantification program using a modified version of GE Q lung.

OBJECTIVE: The value of ventilation-perfusion (VQ) single photon emission tomography/computed tomography (SPECT/CT) lobar quantification for pre-operative assessment of lobectomy and lung volume reduction is known. Our in-house developed software, RAH ventilation perfusion SPECT/CT quantification (RAHVQSQ) has been shown to be able to identify the target lobe for collapse in bronchoscopic lung volume reduction (BLVR) for advanced emphysema. We have proven inter and intra observer reproducibility but are yet to validate the accuracy of our program. This study aims to validate the accuracy of our quantitative program through comparison with a modified version of GE Q lung which is a commercial program certified for clinical use. SUBJECTS AND METHODS: Ventilation-perfusion SPECT/CT data of 19 subjects from our previous study using RAHVQSQ for BLVR assessment were re-analysed using Q lung by 2 technologists independently and in a blinded fashion to determine lobar differential ventilation, perfusion and volume percentages. The data were from GE Hawkeye 4 and external CT, thus a modified version of Q lung was used. To determine interobserver variation in the 3 parameters between the 3 assessors, intraclass correlation coefficient (ICC) and Bland-Altman limits of agreement (LoA) were generated. RESULTS: Paired comparisons between the 3 assessors had high ICC (range for ventilation: 0.69-0.97; perfusion: 0.69-0.97; volume: 0.63-0.97) and means of LoA differences close to zero (range for ventilation: -0.04 - 0.10; perfusion: 0.00-0.02; volume: -0.12 - 0.09) were noted indicative of good concordance for all parameters. CONCLUSION: Using VQ SPECT/CT data of participants with advanced airway disease, our study has found a close concordance of estimated differential lobar ventilation, perfusion and volume percentages using RAHVQSQ when compared with a duplicated blinded assessment using Q lung. The good concordance supports the validity of our quantitative methodology.

Genetic diversity of CHC22 clathrin impacts its function in glucose metabolism.

CHC22 clathrin plays a key role in intracellular membrane traffic of the insulin-responsive glucose transporter GLUT4 in humans. We performed population genetic and phylogenetic analyses of the CHC22-encoding CLTCL1 gene, revealing independent gene loss in at least two vertebrate lineages, after arising from gene duplication. All vertebrates retained the paralogous CLTC gene encoding CHC17 clathrin, which mediates endocytosis. For vertebrates retaining CLTCL1, strong evidence for purifying selection supports CHC22 functionality. All human populations maintained two high frequency CLTCL1 allelic variants, encoding either methionine or valine at position 1316. Functional studies indicated that CHC22-V1316, which is more frequent in farming populations than in hunter-gatherers, has different cellular dynamics than M1316-CHC22 and is less effective at controlling GLUT4 membrane traffic, altering its insulin-regulated response. These analyses suggest that ancestral human dietary change influenced selection of allotypes that affect CHC22's role in metabolism and have potential to differentially influence the human insulin response.