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Aim: Recovery of damaged mucosal surfaces following inflammatory insult requires diverse regenerative mechanisms that remain poorly defined. Previously, we demonstrated that the reparative actions of Trefoil Factor 3 (TFF3) depend upon the enigmatic receptor, leucine rich repeat and immunoglobulin-like domain containing nogo receptor 2 (LINGO2). This study examined the related orphan receptor LINGO3 in the context of intestinal tissue damage to determine whether LINGO family members are generally important for mucosal wound healing and maintenance of the intestinal stem cell (ISC) compartment needed for turnover of mucosal epithelium.Methods and Results: We find that LINGO3 is broadly expressed on human enterocytes and sparsely on discrete cells within the crypt niche, that contains ISCs. Loss of function studies indicate that LINGO3 is involved in recovery of normal intestinal architecture following dextran sodium sulfate (DSS)-induced colitis, and that LINGO3 is needed for therapeutic action of the long acting TFF2 fusion protein (TFF2-Fc), including a number of signaling pathways critical for cell proliferation and wound repair. LINGO3-TFF2 protein-protein interactions were relatively weak however and LINGO3 was only partially responsible for TFF2 induced MAPK signaling suggesting additional un-identified components of a receptor complex. However, deficiency in either TFF2 or LINGO3 abrogated budding/growth of intestinal organoids and reduced expression of the intestinal ISC gene leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5), indicating homologous roles for these proteins in tissue regeneration, possibly via regulation of ISCs in the crypt niche.Conclusion: We propose that LINGO3 serves a previously unappreciated role in promoting mucosal wound healing.

Figure S1. Effect of primary filters on the number of reads. Figure S2. Insert size distribution. Figure S3. Effects of Host DNA on bacterial 16S yield. Figure S4. Total CCS yield vs. DNA yield vs. PCR yields. Figure S5. CCS reads mapping to the human genome from the sinonasal samples. Figure S6. Primer matching filters. Figure S7. Primer matching truncation and nucleotide variability against positive control E.coli forward and reverse primer matches. Figure S8. Primer matching truncation and nucleotide variability against positive control A.tumefaciens forward and reverse primer matches. Figure S9. Histogram of the number of species per DB cluster in NCBI. Figure S10. The effect of PCR cycle number and polymerase choice on OTU abundances in the BEI mock community. Figure S11. The effect of PCR cycles and polymerase on abundances of chimeric molecules in the BEI mock community. Figure S12. Substitution errors in BEI mock community. Figure S13. E.coli MG1655 16S copies and MED analysis. Figure S14. Phylogenetic trees of well---resolved multi---species genera Clostridium and Desulfovibrio. Figure S15. Phylogenetic trees of poorly---resolved multi---species genera Azotobacter and Nonlabens. Figure S16. Phylogenetic trees of multi---species genera with improved species resolution using FL16S for Algoriphagus and Salegentibacter. Figure S17. Phylogeny of AnaerococcusMED nodes from the sinonasal communities plus NCBI database entries. Figure S18. Relationship between species---level confidence in centroid assignments and the number of species in the matching dbOTU. Figure S19. Effective Number of Species as a function of sample read depth. (PDF 3167 kb)

Supplementary Text. (DOCX 56 kb)

The European Position Paper on Rhinosinusitis and Nasal Polyps 2012 is the update of similar evidence based position papers published in 2005 and 2007.The document contains chapters on definitions and classification, we now also proposed definitions for difficult to treat rhinosinusitis, control of disease and better definitions for rhinosinusitis in children. More emphasis is placed on the diagnosis and treatment of acute rhinosinusitis. Throughout the document the terms chronic rhinosinusitis without nasal polyps and chronic rhinosinusitis with nasal polyps are used to further point out differences in pathophysiology and treatment of these two entities. There are extensive chapters on epidemiology and predisposing factors, inflammatory mechanisms, (differential) diagnosis of facial pain, genetics, cystic fibrosis, aspirin exacerbated respiratory disease, immunodeficiencies, allergic fungal rhinosinusitis and the relationship between upper and lower airways. The chapters on paediatric acute and chronic rhinosinusitis are totally rewritten. Last but not least all available evidence for management of acute rhinosinusitis and chronic rhinosinusitis with or without nasal polyps in adults and children is analyzed and presented and management schemes based on the evidence are proposed.