title: A Lactobacillus rhamnosus GG-derived soluble protein, p40, stimulates ligand release from intestinal epithelial cells to transactivate epidermal growth factor receptor.
abstract: p40, a Lactobacillus rhamnosus GG (LGG)-derived soluble protein, ameliorates intestinal injury and colitis, reduces apoptosis, and preserves barrier function by transactivation of the EGF receptor (EGFR) in intestinal epithelial cells. The aim of this study is to determine the mechanisms by which p40 transactivates the EGFR in intestinal epithelial cells. Here we show that p40-conditioned medium activates EGFR in young adult mouse colon epithelial cells and human colonic epithelial cell line, T84 cells. p40 up-regulates a disintegrin and metalloproteinase domain-containing protein 17 (ADAM17) catalytic activity, and broad spectrum metalloproteinase inhibitors block EGFR transactivation by p40 in these two cell lines. In ADAM17-deficient mouse colonic epithelial (ADAM17(-/-) MCE) cells, p40 transactivation of EGFR is blocked, but can be rescued by re-expression with WT ADAM17. Furthermore, p40 stimulates release of heparin binding (HB)-EGF, but not transforming growth factor (TGF)α or amphiregulin, in young adult mouse colon cells and ADAM17(-/-) MCE cells overexpressing WT ADAM17. Knockdown of HB-EGF expression by siRNA suppresses p40 effects on transactivating EGFR and Akt, preventing apoptosis, and preserving tight junction function. The effects of p40 on HB-EGF release and ADAM17 activation in vivo are examined after administration of p40-containing pectin/zein hydrogel beads to mice. p40 stimulates ADAM17 activity and EGFR activation in colonic epithelial cells and increases HB-EGF levels in blood from WT mice, but not from mice with intestinal epithelial cell-specific ADAM17 deletion. Thus, these data define a mechanism of a probiotic-derived soluble protein in modulating intestinal epithelial cell homeostasis through ADAM17-mediated HB-EGF release, leading to transactivation of EGFR.
authors.affiliation: From the Departments of Pediatrics and.
authors.forename: Fang
authors.initials: F
authors.lastname: Yan
authors.affiliation: From the Departments of Pediatrics and.
authors.forename: Liping
authors.initials: L
authors.lastname: Liu
authors.affiliation: the Departments of Pediatrics and Communicable Diseases and; Molecular and Integrative Physiology, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Dempsey
authors.affiliation: the Departments of Pediatrics and Communicable Diseases and; Molecular and Integrative Physiology, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Yu-Hwai
authors.initials: YH
authors.lastname: Tsai
authors.affiliation: the Department of Pathology, University of Washington School of Medicine, Seattle, Washington 98195.
authors.forename: Elaine W
authors.initials: EW
authors.lastname: Raines
authors.affiliation: the Department of Pathology, University of Washington School of Medicine, Seattle, Washington 98195.
authors.forename: Carole L
authors.initials: CL
authors.lastname: Wilson
authors.affiliation: From the Departments of Pediatrics and; the Department of Gastroenterology, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Hailong
authors.initials: H
authors.lastname: Cao
authors.affiliation: Medicine, Division of Gastroenterology, Hepatology and Nutrition, Vanderbilt University Medical Center, Nashville, Tennessee 37232.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Cao
authors.affiliation: the Eastern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, Pennsylvania 19038, and.
authors.forename: LinShu
authors.initials: L
authors.lastname: Liu
authors.affiliation: the Departments of Pediatrics and Biochemistry and Molecular Biology, University of Southern California and Saban Research Institute of Children's Hospital Los Angeles, Los Angeles, California 90089. Electronic address: fang.yan@vanderbilt.edu.
authors.forename: D Brent
authors.initials: DB
authors.lastname: Polk
mesh_terms: D051722:ADAM Proteins / Q000096:biosynthesis / Q000235:genetics; D000072198:ADAM17 Protein; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D004789:Enzyme Activation / Q000235:genetics; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology; D066246:ErbB Receptors / Q000096:biosynthesis* / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D055785:Gene Knockdown Techniques; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology; D052201:Lacticaseibacillus rhamnosus / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D019936:Probiotics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D016211:Transforming Growth Factor alpha / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D016211:Transforming Growth Factor alpha; C560306:p40 protein, Lactobacillus rhamnosus; C512478:EGFR protein, human; C541485:EGFR protein, mouse; D066246:ErbB Receptors; D051057:Proto-Oncogene Proteins c-akt; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human; C000606463:Adam17 protein, mouse
keywords: Apoptosis; Epidermal Growth Factor (EGF); Intestinal Epithelium; Probiotics; Tight Junctions
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