title: Application of Minicircle Technology of Self-Reproducing Synthetic Protein Drugs in Preventing Skin Allograft Rejection.
abstract: BACKGROUND
  Recently, it has been reported that minicircle vectors could allow the expression of transgenes using the protein synthesis system of the host. Here, we tested a novel strategy to permit the production of synthetic biologics using minicircle technology and evaluated their feasibility as a therapeutic tool in a skin allograft model.
  
  
  METHODS
  We engineered vectors to carry cassette sequences for tocilizumab [anti-soluble interleukin-6 receptor (sIL-6R) antibody] and/or etanercept [tumor necrosis factor receptor 2 (TNFR2)-Fc fusion protein], and then isolated minicircle vectors from the parent vectors. We verified the production of proteins from minicircles and their duration in HEK293T cells and mice. We also evaluated whether these proteins were expressed at levels sufficient to ameliorate skin allograft rejection in mice.
  
  
  RESULTS
  Each minicircle transfected into cells was detectable for at least 30 days. In mice, the drugs were mainly expressed in the liver and were detectable for at least 10 days after a single injection. These drugs were also detected in the blood. Treatment of mice with minicircles prolonged skin allograft survival, which was accompanied by a reduction of the number of interferon-γ+ or interleukin-17+ lymphocytes and an induction of forkhead box P3 expression. These findings suggest that blocking of sIL-6R and/or TNF-α using minicircles encoding tocilizumab and/or etanercept was functionally active and relevant for preventing acute allograft rejection.
  
  
  CONCLUSIONS
  Self-reproducing synthetic protein drugs produced using minicircle technology are potentially powerful tools for preventing acute rejection in transplantation.
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Sun Woo
authors.initials: SW
authors.lastname: Lim
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Young Kyun
authors.initials: YK
authors.lastname: Kim
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Narae
authors.initials: N
authors.lastname: Park
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Long
authors.initials: L
authors.lastname: Jin
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Jian
authors.initials: J
authors.lastname: Jin
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Kyoung Chan
authors.initials: KC
authors.lastname: Doh
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Ji Hyeon
authors.initials: JH
authors.lastname: Ju
authors.affiliation: Convergent Research Consortium for Immunologic Disease, Seoul St, Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
authors.forename: Chul Woo
authors.initials: CW
authors.lastname: Yang
mesh_terms: D064591:Allografts; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000627:therapeutic use; D000068800:Etanercept / Q000627:therapeutic use; D006084:Graft Rejection / Q000276:immunology / Q000517:prevention & control*; D006085:Graft Survival / Q000276:immunology*; D057809:HEK293 Cells; D006801:Humans; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D051379:Mice; D016038:Skin Transplantation / Q000379:methods*
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D007166:Immunosuppressive Agents; C502936:tocilizumab; D000068800:Etanercept