title: Nanoparticles with surface antibody against CD98 and carrying CD98 small interfering RNA reduce colitis in mice.
abstract: OBJECTIVE
  Nanoparticles have been explored as carriers of small interfering RNAs (siRNAs) and might be developed to treat patients with inflammatory bowel disease (IBD). Overexpression of CD98 on the surface of colonic epithelial cells and macrophages promotes the development and progression of IBD. We developed an orally delivered hydrogel that releases nanoparticles with single-chain CD98 antibodies on their surface (scCD98 functionalized) and loaded with CD98 siRNA (siCD98). We tested the ability of the nanoparticles to reduce levels of CD98 in the colons of mice with colitis.
  
  
  METHODS
  scCD98-functionalized siCD98-loaded nanoparticles were fabricated using a complex coacervation technique. We investigated the cellular uptake and lysosome escape profiles of the nanoparticles in Colon-26 cells and RAW 264.7 macrophages using fluorescence microscopy. Colitis was induced by transfer of CD4(+)CD45RB(high) T cells to Rag(-/-) mice or administration of dextran sodium sulfate to C57BL/6 mice. Mice were then given hydrogel (chitosan and alginate) containing scCD98-functionalized nanoparticles loaded with siCD98 or scrambled siRNA (control) via gavage.
  
  
  RESULTS
  The scCD98-functionalized nanoparticles were approximately 200 nm in size and had high affinity for CD98-overexpressing cells. The scCD98-functionalized siCD98-loaded nanoparticles significantly reduced levels of CD98 in Colon-26 cells and RAW 264.7 macrophages, along with production of inflammatory cytokines (tumor necrosis factor α, interleukin-6, and interleukin-12). In mice with colitis, administration of the scCD98-functionalized siCD98-loaded nanoparticles reduced colon expression of CD98. Importantly, the severity of colitis was also reduced compared with controls (based on loss of body weight, myeloperoxidase activity, inflammatory cytokine production, and histological analysis). Approximately 24.1% of colonic macrophages (CD11b(+)CD11c(-)F4/80(+)) in the mice had taken up fluorescently labeled siRNA-loaded nanoparticles within 12 hours of administration.
  
  
  CONCLUSIONS
  Nanoparticles containing surface CD98 antibody and loaded with siCD98 reduce expression of this protein by colonic epithelial cells and macrophages, and oral administration decreases the severity of colitis in mice. This nanoparticle in hydrogel (chitosan/alginate) formulation might be developed to treat patients with IBD.
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta. Electronic address: bxiao@gsu.edu.
authors: Bo
authors: B
authors: Xiao
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta.
authors: Hamed
authors: H
authors: Laroui
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta; Atlanta Veterans Affairs Medical Center, Decatur.
authors: Emilie
authors: E
authors: Viennois
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta.
authors: Saravanan
authors: S
authors: Ayyadurai
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta.
authors: Moiz A
authors: MA
authors: Charania
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta.
authors: Yuchen
authors: Y
authors: Zhang
authors: Center for Inflammation, Immunity and Infection, Department of Biology, Georgia State University, Atlanta.
authors: Zhan
authors: Z
authors: Zhang
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta.
authors: Mark T
authors: MT
authors: Baker
authors: Center for Inflammation, Immunity and Infection, Department of Biology, Georgia State University, Atlanta.
authors: Benyue
authors: B
authors: Zhang
authors: Center for Inflammation, Immunity and Infection, Department of Biology, Georgia State University, Atlanta; Department of Pathology, School of Medicine, Emory University, Atlanta, Georgia.
authors: Andrew T
authors: AT
authors: Gewirtz
authors: Center for Diagnostics and Therapeutics, Institute for Biomedical Science, Departments of Biology and Chemistry, Georgia State University, Atlanta; Atlanta Veterans Affairs Medical Center, Decatur.
authors: Didier
authors: D
authors: Merlin
mesh_terms: D000284:Administration, Oral; D000464:Alginates / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry; D003092:Colitis / Q000139:chemically induced / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D003106:Colon / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D016264:Dextran Sulfate; D004195:Disease Models, Animal; D027261:Fusion Regulatory Protein-1 / Q000235:genetics* / Q000276:immunology*; D015316:Genetic Therapy / Q000379:methods*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D020100:Hydrogels; D018664:Interleukin-12 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D050997:Nanomedicine / Q000379:methods*; D053758:Nanoparticles; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000378:metabolism; D012720:Severity of Illness Index; D057127:Single-Chain Antibodies / Q000008:administration & dosage*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000464:Alginates; D027261:Fusion Regulatory Protein-1; D006603:Hexuronic Acids; D018398:Homeodomain Proteins; D020100:Hydrogels; D015850:Interleukin-6; D034741:RNA, Small Interfering; D057127:Single-Chain Antibodies; D014409:Tumor Necrosis Factor-alpha; C064658:RAG-1 protein; D018664:Interleukin-12; D020723:Glucuronic Acid; D048271:Chitosan; D016264:Dextran Sulfate
keywords: CD98; Colitis-Targeted Therapy; Gene Silencing; Mouse Model