RESEARCH ARTICLE

Adenovirus-mediated antisense ERK2 gene therapy ameliorates chronic allograft nephropathy in a rat model

  • Zhao DING ,
  • Zhishui CHEN ,
  • Xilin CHEN ,
  • Ming CAI ,
  • Hui GUO ,
  • Nianqiao GONG
Expand
  • Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

Received date: 05 Jan 2009

Accepted date: 01 Mar 2009

Published date: 05 Jun 2009

Copyright

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg

Abstract

To investigate the effect and underlying mechanism of adenovirus-mediated antisense ERK2 (Adanti-ERK2) gene therapy upon chronic allograft nephropathy (CAN) of rats, male Lewis (LEW, RT11) rats received male Fisher (F344, RT11v1) renal allografts. The recipients were divided into three groups: (1) empty control group; (2) vector control group; (3) gene therapy group. All recipients were sacrificed for the grafts and serum analysis at the 24th week after transplantation. Morphometric analysis was used to determine the fibrosis of grafts. Immunohistochemistry was used to detect the expression of E-Cadherin, Vimentin, TβR I and the infiltration of CD4+ T lymphocyte, CD8+ T lymphocyte and ED-1+ monocytes. Enzyme linked immunosorbent assay (ELISA) was used to detect TGF-β1 in serum. The grafts in the control group and vector control group showed CAN. There was less E-Cadherin in renal tubular epithelial cells in the empty control group but more Vimentin and TβR I. In the gene therapy group, the fibrosis was ameliorated and fewer T lymphocytes and ED-1+ monocytes infiltrated in the interstitium. There was no significant difference in the expression of E-Cadherin between the gene therapy group and normal rats. Compared with the empty control group, the expression of TGF-β1 in the gene therapy group was down-regulated. Adanti-ERK2 gene therapy protects the renal allograft and attenuates graft fibrosis, which may be correlated with a decreased renal tubular epithelial mesenchymal transition, a decreased infiltration of CD4+ T lymphocyte, CD8+ T lymphocytes and ED-1+ monocytes in renal interstitium, and the down-regulated TGF-β1 expression.

Cite this article

Zhao DING , Zhishui CHEN , Xilin CHEN , Ming CAI , Hui GUO , Nianqiao GONG . Adenovirus-mediated antisense ERK2 gene therapy ameliorates chronic allograft nephropathy in a rat model[J]. Frontiers of Medicine, 2009 , 3(2) : 204 -210 . DOI: 10.1007/s11684-009-0039-0

Acknowledgements

The study was supported by the National Natural Science Foundation of China (Grant No. 30300324).
1
PascualM, TheruvathT, KawaiT, Tolkoff-RubinN, CosimiA B. Strategies to improve long-term outcomes after renal transplantation. N Engl J Med, 2002, 346(8): 580–590

DOI

2
BurnsW C, KantharidisP, ThomasM C. The role of tubular epithelial-mesenchymal transition in progressive kidney disease. Cells Tissues Organs, 2007, 185(1-3): 222–31

DOI

3
NankivellB J, BorrowsR J, FungC L, O'ConnellP J, AllenR D, Chapman J R. The natural history of chronic allograft nephropathy. N Engl J Med, 2003, 349(24): 2326–2333

DOI

4
KyriakisJ M, AvruchJ. Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev, 2001, 81(2): 807–869

5
BostF, AouadiM, CaronL, EvenP, BelmonteN, ProtM, DaniC, HofmanP, PagèsG, PouysségurJ, Le Marchand-BrustelY, BinétruyB. The extracellular signal–regulated kinase isoform ERK1 is specifically required for in vitro and in vivo adipogenesis. Diabetes, 2005, 54(2): 402–411

DOI

6
VantaggiatoC, FormentiniI, BondanzaA, BoniniC, NaldiniL, BrambillaR. ERK1 and ERK2 mitogen-activated protein kinases affect Ras-dependent cell signaling differentially. J Biol, 2006, 5(5): 14

7
MizuguchiH, KayM A. A simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors. Hum Gene Ther, 1999, 10(12): 2013–2017

DOI

8
LeeS. An improved technique of renal transplantation in the rat. Surgery, 1967, 61(5): 771–773

9
PascualM, TheruvathT, KawaiT, Tolkoff-RubinN, CosimiA B. Strategies to improve long-term outcomes after renal transplantation. N Engl J Med, 2002, 346(8): 580–590

DOI

10
IwanoM, PliethD, DanoffT M, XueC, OkadaH, NeilsonE G. Evidence that fibroblasts derive from epithelium during tissue fibrosis. J Clin Invest, 2002, 110(3): 341–350

11
GongN, ZhangW, LiG, GuoH, YeQ. MAPK cascade pathway and anti-stress response of hepatocyte after live transplantation. Chin J Organ Transplant, 2002, 23(5): 288–290 (in Chinese)

12
LiG, GongN, YeQ, GuoH. Different effects of several signal pathways after liver transplantation. Zhonghua Ganzangbing Zazhi, 2003, 11(12): 742–745 (in Chinese).

13
DongC, GongN, ChenZ, ChenX, XuQ, GuoH, ZengZ, MingC, ChenZ K. Antisense ERK1/2 oligodeoxynucleotide gene therapy attenuates graft arteriosclerosis of aortic transplant in a rat model. Transplant Proc, 2006, 38(10): 3304–3306

DOI

14
WhitehurstA W, RobinsonF L, MooreM S, CobbM H. The death effector domain protein PEA-15 prevents nuclear entry of ERK2 by inhibiting required interactions. J Biol Chem, 2004, 279(13): 12840–12847

DOI

15
ChenR H, AbateC, BlenisJ. Phosphorylation of the c-Fos transrepression domain by mitogen-activated protein kinase and 90-kDa ribosomal S6 kinase. Proc Natl Acad Sci USA, 1993, 90(23): 10952–10956

DOI

16
GongN, PleyerU, VogtK, AnegonI, FlügelA, VolkH D, RitterT. Local overexpression of nerve growth factor in rat corneal transplants improves allograft survival. Invest Ophthalmol Vis Sci, 2007, 48(3): 1043–1052

DOI

17
TilneyN L, WhitleyW D, TulliusS G, HeemannU W, WasowskaB, BaldwinW M 3rd, HancockW W. Serial analysis of cytokines, adhesion molecule expression, and humoral responses during development of chronic kidney allograft rejection in a new rat model. Transplant Proc, 1993, 25(1 Pt 2): 861–862

18
TulliusS G, HancockW W, HeemannU, AzumaH, TilneyN L. Reversibility of chronic renal allograft rejection. Critical effect of time after transplantation suggests both host immune dependent and independent phases of progressive injury. Transplantation, 1994, 58(1): 93–99

19
Meindl-BeinkerN M, DooleyS. Transforming growth factor-beta and hepatocyte transdifferentiation in liver fibrogenesis. J Gastroenterol Hepatol, 2008, 23(Suppl 1): S122–S127

DOI

20
RobertsonH, AliS, McDonnellB J, BurtA D, KirbyJ A. Chronic renal allograft dysfunction: The role of T cell-mediated tubular epithelial to mesenchymal cell transition. J Am Soc Nephrol, 2004, 15(2): 390–397

DOI

21
ReedR C, BerwinB, BakerJ P, NicchittaC V. GRP94/gp96 elicits ERK activation in murine macrophages. A role for endotoxin contamination in NF-kappa B activation and nitric oxide production. J Biol Chem, 2003, 278(34): 31853–31860

DOI

22
ZhangY L, DongC. MAP kinases in immune responses. Cell Mol Immunol, 2005, 2(1): 20–27

Outlines

/