title: Intracellular delivery of desulfated heparin with bile acid conjugation alleviates T cell-mediated inflammatory arthritis via inhibition of RhoA-dependent transcellular diapedesis.
abstract: Heparin has a potential regulatory role in inflammatory diseases. However, the anticoagulant activity and poor oral bioavailability of heparin limit its use as an anti-inflammatory agent. Conjugation of bis-deoxycholic acid to 6-O-desulfated low molecular weight heparin (6DSHbD) was efficiently internalized by activated endothelial cells via a 2-step model, in which heparin attaches to adhesion molecules that facilitate accessibility of the bile acid conjugate to membrane transporters. The critical role of P-selectin during endothelial cell uptake of 6DSHbD by arthritic tissue was confirmed in p-selectin(-/-) arthritic mice. Intracellular 6DSHbD inhibited transcellular diapedesis of T cells through activated endothelial cells and impaired both the formation of ICAM-1-rich docking structures at the T cell contact surface and subsequent cytoskeletal rearrangement. Furthermore, 6DSHbD blocked activation of RhoA-GTPase and phosphorylation of ezrin/radixin/moesin induced by ICAM-1 cross-linking on activated endothelial cells, thereby impairing lymphocyte transcellular transmigration. After oral administration 6DSHbD was preferentially delivered to inflamed joint tissue, particularly in and around post-capillary venular endothelium and inhibited effector T cell homing to arthritic joints. Aggravation of collagen-induced arthritis conferred by the transfer of effector T cells was suppressed by oral 6DSHbD. Thus, intracellular heparin exerts anti-inflammatory effects through the inhibition of RhoA-dependent transendothelial recruitment of T cells and may have applications in the treatment of chronic inflammatory arthritis.
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: Jin Hee
authors.initials: JH
authors.lastname: Kang
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Seung Rim
authors.initials: SR
authors.lastname: Hwang
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: Shijin
authors.initials: S
authors.lastname: Sung
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: Ji Ae
authors.initials: JA
authors.lastname: Jang
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: Md Mahmudul
authors.initials: MM
authors.lastname: Alam
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: Keum Hee
authors.initials: KH
authors.lastname: Sa
authors.affiliation: Department of Biochemistry and Cellular Biology, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: Sang-Yeob
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Department of Biochemistry and Cellular Biology, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea; Cell and Matrix Research Institute, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea.
authors.forename: In San
authors.initials: IS
authors.lastname: Kim
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Young Ro
authors.initials: YR
authors.lastname: Byun
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea; Department of Biochemistry and Cellular Biology, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea; Cell and Matrix Research Institute, Kyungpook National University School of Medicine, Daegu 700-422, Republic of Korea. Electronic address: ymkang@knu.ac.kr.
authors.forename: Young Mo
authors.initials: YM
authors.lastname: Kang
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000627:therapeutic use*; D001172:Arthritis, Rheumatoid / Q000188:drug therapy* / Q000276:immunology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003840:Deoxycholic Acid / Q000737:chemistry*; D004195:Disease Models, Animal; D016503:Drug Delivery Systems; D006493:Heparin / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000627:therapeutic use; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology; D008811:Mice, Inbred DBA; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D057705:Transendothelial and Transepithelial Migration / Q000187:drug effects* / Q000276:immunology; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; C410523:heparin, O-desulfated; D003840:Deoxycholic Acid; D018799:Intercellular Adhesion Molecule-1; D006493:Heparin; D020742:rhoA GTP-Binding Protein
keywords: Bile acid conjugation; Desulfation; Endothelial cell; Inflammatory arthritis; Low molecular weight heparin; T cell