The core planar cell polarity gene, Vangl2, directs adult corneal epithelial cell alignment and migration

This work was supported by a Biotechnology and Biological Sciences Research Council (BBSRC) DTG PhD studentship to A.F., an Anatomical Society PhD Studentship (‘The Roles of planar cell polarity genes in a classical anatomical system: the cornea’) to D.A.P./J.M.C. and BBSRC Project Grants BB/J015172/1 and BB/J015237/1 to J.D.W. and J.M.C., respectively.Peer reviewedPublisher PD

Dataset for One-pot assembly of a biocompatible triazole-linked gene by click-DNA ligation.

Data supporting Kukwikila, Nsimba M., Gale, N., El-Sagheer, A.H., Brown, Tom and Tavassoli, Ali (2017) Assembly of a biocompatible triazole-linked gene by one-pot click-DNA ligation Nature Chemistry</span

The flagellin of candidate live biotherapeutic Enterococcus gallinarum MRx0518 is a potent immunostimulant

Abstract Many links between gut microbiota and disease development have been established in recent years, with particular bacterial strains emerging as potential therapeutics rather than causative agents. In this study we describe the immunostimulatory properties of Enterococcus gallinarum MRx0518, a candidate live biotherapeutic with proven anti-tumorigenic efficacy. Here we demonstrate that strain MRx0518 elicits a strong pro-inflammatory response in key components of the innate immune system but also in intestinal epithelial cells. Using a flagellin knock-out derivative and purified recombinant protein, MRx0518 flagellin was shown to be a TLR5 and NF-κB activator in reporter cells and an inducer of IL-8 production by HT29-MTX cells. E. gallinarum flagellin proteins display a high level of sequence diversity and the flagellin produced by MRx0518 was shown to be more potent than flagellin from E. gallinarum DSM100110. Collectively, these data infer that flagellin may play a role in the therapeutic properties of E. gallinarum MRx0518

Word document with supplementary figures and details of primers and siRNAs. from The core planar cell polarity gene, <i>Vangl2</i>, directs adult corneal epithelial cell alignment and migration

This study shows that the core planar cell polarity (PCP) genes direct the aligned cell migration in the adult corneal epithelium, a stratified squamous epithelium on the outer surface of the vertebrate eye. Expression of multiple core PCP genes was demonstrated in the adult corneal epithelium. PCP components were manipulated genetically and pharmacologically in human and mouse corneal epithelial cells <i>in vivo</i> and <i>in vitro</i>. Knockdown of <i>VANGL2</i> reduced the directional component of migration of human corneal epithelial (HCE) cells without affecting speed. It was shown that signalling through PCP mediators, dishevelled, dishevelled-associated activator of morphogenesis and Rho-associated protein kinase directs the alignment of HCEs by affecting cytoskeletal reorganization. Cells in which <i>VANGL2</i> were disrupted tended to misalign on grooved surfaces and migrate across, rather than parallel to the grooves. Adult corneal epithelial cells in which <i>Vangl2</i> had been conditionally deleted showed a reduced rate of wound-healing migration. Conditional deletion of <i>Vangl2</i> in the mouse corneal epithelium ablated the normal highly stereotyped patterns of centripetal cell migration <i>in vivo</i> from the periphery (limbus) to the centre of the cornea. Corneal opacity owing to chronic wounding is a major cause of degenerative blindness across the world, and this study shows that Vangl2 activity is required for directional corneal epithelial migration