TFF3 interacts with LINGO2 to regulate EGFR activation for protection against colitis and gastrointestinal helminths

September 27, 2019

Intro

Nature Communications volume 10, Article number: 4408 (2019) https://doi.org/10.1038/s41467-019-12315-1

Abstract

: Intestinal epithelial cells (IEC) have important functions in nutrient absorption, barrier integrity, regeneration, pathogen-sensing, and mucus secretion. Goblet cells are a specialized cell type of IEC that secrete Trefoil factor 3 (TFF3) to regulate mucus viscosity and wound healing, but whether TFF3-responsiveness requires a receptor is unclear. We postulated that TFF3 interacted with its receptor through low-affinity interactions reliant upon carbohydrates because TFF3 glycosylation has been shown critical for biological activity34,35, therefore U937 cells were subjected to the TRICEPSTM protocol as a biochemical screening strategy to identify glycosylated transmembrane protein(s) on the cell surface32,36. As bait, rhTFF3 was covalently linked to the TRICEPS probe and incubated with PMA-treated U937 that had been treated with sodium periodate. Cell pellets were subjected to glycosidase digestion and peptides generated using mass spectrometry. As a positive control to account for enrichment efficiency, recombinant human insulin was used as bait for the insulin receptor. Whereas the TFF3-probe induced an 8-fold enrichment of LINGO2 (LIGO2) peptide, the insulin-probe induced a 7.76-fold-enrichment in INSR peptide (Fig. 1b). No other enriched peptides in this screen met this level of enrichment or were derived from proteins that satisfied our selection criterion (e.g., extracellular domain, transmembrane region, cytoplasmic tail).Here, we show that leucine rich repeat receptor and nogo-interacting protein 2 (LINGO2) is essential for TFF3-mediated functions. LINGO2 immunoprecipitates with TFF3, co-localizes with TFF3 on the cell membrane of IEC, and allows TFF3 to block apoptosis. We further show that TFF3-LINGO2 interactions disrupt EGFR-LINGO2 complexes resulting in enhanced EGFR signaling. Excessive basal EGFR activation in Lingo2 deficient mice increases disease severity during colitis and augments immunity against helminth infection. Conversely, TFF3 deficiency reduces helminth immunity. Thus, TFF3-LINGO2 interactions de-repress inhibitory LINGO2-EGFR complexes, allowing TFF3 to drive wound healing and immunity.

Author

Nicole Maloney Belle1, Yingbiao Ji1,5, Karl Herbine1,5, Yun Wei2,3,5, JoonHyung Park1,5, Kelly Zullo1, Li-Yin Hung1,2, Sriram Srivatsa1, Tanner Young1, Taylor Oniskey2, Christopher Pastore1, Wildaliz Nieves4, Ma Somsouk4 & De’Broski R. Herbert1,2 1 Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA 19140, USA. 2 Division of Experimental Medicine, University of California, San Francisco, San Francisco, CA 94110, USA. 3 Department of Inflammation and Oncology, Amgen Inc., 1120 Veterans Boulevard, South San Francisco, CA 94080, USA. 4 Division of Gastroenterology at ZSFG, University of California, San Francisco, San Francisco, CA 94110, USA. 5These authors contributed equally: Yingbiao Ji, Karl Herbine, Yun Wei, JoonHyung Park. *email: debroski@vet.upenn.edu