title: Spherical nucleic acid targeting microRNA-99b enhances intestinal MFG-E8 gene expression and restores enterocyte migration in lipopolysaccharide-induced septic mice.
abstract: Milk fat globule-EGF factor 8 (MFG-E8) maintains the intestinal homeostasis by enhancing enterocyte migration and attenuating inflammation. We previously reported that sepsis is associated with down-regulation of intestinal MFG-E8 and impairment of enterocyte migration. Here, we showed that impairment of intestinal epithelial cell migration occurred in lipopolysaccharide (LPS)-induced septic mice. Treatment of RAW264.7 cells (a murine macrophage-like cell line) with LPS increased expression of miR-99b, a microRNA that is predicted to target mouse MFG-E8 3'UTR. Using a luciferase assay, we showed that miR-99b mimic suppressed the activity of a reporter containing MFG-E8 3'UTR. This suggests the role of miR-99b in inhibition of MFG-E8 gene expression. In addition, we developed an anti-miR99b spherical nucleic acid nanoparticle conjugate (SNA-NC(anti-miR99b)). Treatment of both naïve and LPS-challenged cells with SNA-NC(anti-miR99b) enhanced MFG-E8 expression in the cells. Administration of SNA-NC(anti-miR99b) rescued intestinal MFG-E8 expression in LPS-induced septic mice and attenuated LPS inhibitory effects on intestinal epithelial cell migration along the crypt-villus axis. Collectively, our study suggests that LPS represses MFG-E8 expression and disrupts enterocyte migration via a miR-99b dependent mechanism. Furthermore, this work shows that SNA-NC(anti-miR99b) is a novel nanoparticle-conjugate capable of rescuing MFG-E8 gene expression and maintaining intestinal epithelial homeostasis in sepsis.
authors.affiliation: Center for Intestinal and Liver Inflammation Research, Stanley Manne Children's Research Institute, Ann &Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.|Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Chemistry and the International Institute for Nanotechnology, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Hao
authors.affiliation: Center for Intestinal and Liver Inflammation Research, Stanley Manne Children's Research Institute, Ann &Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.|Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Heng-Fu
authors.initials: HF
authors.lastname: Bu
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Scott
authors.affiliation: Center for Intestinal and Liver Inflammation Research, Stanley Manne Children's Research Institute, Ann &Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.|Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.|Department of Gastroenterology, Peking University People's Hospital, Beijing 100044, P. R. China.
authors.forename: Ke
authors.initials: K
authors.lastname: Tian
authors.affiliation: Center for Intestinal and Liver Inflammation Research, Stanley Manne Children's Research Institute, Ann &Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.|Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Fangyi
authors.initials: F
authors.lastname: Liu
authors.affiliation: Center for Intestinal and Liver Inflammation Research, Stanley Manne Children's Research Institute, Ann &Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.|Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Isabelle G
authors.initials: IG
authors.lastname: De Plaen
authors.affiliation: Department of Gastroenterology, Peking University People's Hospital, Beijing 100044, P. R. China.
authors.forename: Yulan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Chemistry and the International Institute for Nanotechnology, Northwestern University, Evanston, IL 60208, USA.|Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Chad A
authors.initials: CA
authors.lastname: Mirkin
authors.affiliation: Center for Intestinal and Liver Inflammation Research, Stanley Manne Children's Research Institute, Ann &Robert H. Lurie Children's Hospital of Chicago, Chicago, IL 60611, USA.|Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.|Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Xiao-Di
authors.initials: XD
authors.lastname: Tan
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis* / Q000235:genetics; D002465:Cell Movement / Q000187:drug effects* / Q000235:genetics; D020895:Enterocytes / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D018390:Gene Targeting; D008070:Lipopolysaccharides / Q000633:toxicity*; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008894:Milk Proteins / Q000096:biosynthesis* / Q000235:genetics; D000067996:RAW 264.7 Cells; D018805:Sepsis / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology
chemical_list: D000954:Antigens, Surface; D008070:Lipopolysaccharides; C490919:Mfge8 protein, mouse; D035683:MicroRNAs; D008894:Milk Proteins; C573158:Mirn99 microRNA, mouse