Encapsulating peritoneal sclerosis (EPS) is a devastating fibrotic complication in patients treated with peritoneal dialysis (PD). Transforming growth factor β1 (TGF-β1) is a pivotal factor in the induction of EPS.
Methods
To develop pyrrole-imidazole (PI) polyamide, a novel gene silencer, targeted to the TGF-β1 promoter (Polyamide) for EPS, we examined the effects of Polyamide on messenger RNA (mRNA) expression of TGF-β 1, vascular endothelial growth factor (VEGF), and extracellular matrix (ECM) in mesothelial cells in vitro, and on the thickness of injured peritoneum evaluated by histology and high- resolution regional elasticity mapping in rats in vivo.
Results
Polyamide significantly lowered mRNA expression of TGF-β 1 and ECM in vitro. Polyamide labeled with fluorescein isothiocyanate was taken up into the injured peritoneum and was strongly localized in the nuclei of most cells. Polyamide 1 mg was injected intraperitoneally 1 or 3 times in rats receiving a daily intraperitoneal injection of chlorhexidine gluconate and ethanol (CHX) for 14 days. Polyamide significantly suppressed peritoneal thickening and the abundance of TGF-β 1 and fibronectin mRNA, but did not affect expression of VEGF mRNA in the injured peritoneum. Elasticity distribution mapping showed that average elasticity was significantly lower in Polyamide-treated rats than in rats treated solely with CHX.
Conclusions
Polyamide suppressed the stiffness, ECM formation, and thickening of the injured peritoneum that occurs during EPS pathogenesis. These data suggest that PI polyamide targeted to the TGF-β 1 promoter will be a specific and feasible therapeutic strategy for patients with EPS.
HiramatsuM.Japanese Society for Elderly Patients on Peritoneal Dialysis. How to improve survival in geriatric peritoneal dialysis patients.Perit Dial Int2007; 27(Suppl 2): S185–9.
2.
KawaguchiY., KawanishiH., MujaisS., TopleyN., OreopoulosD.G.Encapsulating peritoneal sclerosis: definition, etiology, diagnosis, and treatment. International Society for Peritoneal Dialysis Ad Hoc Committee on Ultrafiltration Management in Peritoneal Dialysis.Perit Dial Int2000; 20(Suppl 4) S43–55.
3.
KredietR.T.The peritoneal membrane in chronic peritoneal dialysis.Kidney Int1999; 55: 341–56.
IgnotzR.A., MassaguéJ.. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix.J Biol Chem1986; 261: 4337–45.
6.
LiuL., ShiC.X., GhayurA., ZhangC., SuJ.Y., HoffC.M.Prolonged peritoneal gene expression using a helper-dependent adenovirus.Perit Dial Int2009; 29: 508–16.
7.
Yáñez–MóM., Lara-PezziE., SelgasR., Ramírez-HuescaM., Domínguez-JiménezC., Jiménez-HeffernanJ.A.Peritoneal dialysis and epithelial-to-mesenchymal transition of mesothelial cells.N Eng J Med2003; 348: 403–13.
8.
IoH., HamadaC., RoY., ItoY., HiraharaI., TominoY.Morphologic changes of peritoneum and expression of VEGF in encapsulated peritoneal sclerosis rat models.Kidney Int2004; 65: 1927–36.
9.
MatsuokaT., MaedaY., MatsuoK., NaikiY., TamaiY., SakaguchiM.Hepatocyte growth factor prevents peritoneal fibrosis in an animal model of encapsulating peritoneal sclerosis.J Nephrol2008; 21: 64–73.
10.
TraugerJ.W., BairdE.E., DervanP.B.Recognition of DNA by designed ligands at subnanomolar concentrations.Nature1996; 382: 559–61.
11.
WhiteS., BairdE.E., DervanP.B.On the pairing rules for recognition in the minor groove of DNA by pyrrole–imidazole polyamides.Chem Biol1997; 4: 569–78.
12.
MurtyM.S., SugiyamaH.Biology of N-methylpyrrole-N-methylimidazole hairpin polyamide.Biol Pharm Bull2004; 27: 468–74.
13.
GottesfeldJ.M., NeelyL., TraugerJ.W., BairdE.E., DervanP.B.Regulation of gene expression by small molecules.Nature1997; 387: 202–5.
14.
FujimotoK., IidaH., KawakamiM., BandoT., TaoZ.F., SugiyamaH.Sequence-specific protection of plasmid DNA from restriction endonuclease hydrolysis by pyrrole–imidazole– cyclopropapyrroloindole conjugates.Nucleic Acids Res2002; 30: 3748–53.
15.
MurayamaY., MizunoJ., KamakuraH., FuetaY., NakamuraH., AkaishiK.Mouse zona pellucida dynamically changes its elasticity during oocyte maturation, fertilization and early embryo development.Hum Cell2006; 19: 119–25.
16.
MurayamaY., ConstantinouC.E., OmataS.Micro-mechanical sensing platform for the characterization of the elastic properties of the ovum via uniaxial measurement.J Biomech2004; 37: 67–72.
MurayamaY., HarutaM., HatakeyamaY., ShiinaT., SakumaH., TakenoshitaS.Development of a new instrument for examination of stiffness in the breast using haptic sensor technology.Sensors Actuators A Phys2008; 143: 430–8.
19.
MurayamaY., ConstantinouC.E., OmataS.Development of tactile mapping system for the stiffness characterization of tissue slice using novel tactile sensing technology.Sensors Actuators A Phys2005; 120: 543–9.
20.
OieT., SuzukiH., MurayamaY., FukudaT., OmataS., KandaK.Surface elasticity imaging of vascular tissues in liquid environment by scanning haptic microscope.J Artif Organs2010; 13: 121–5.
21.
LaiY.M., FukudaN., UenoT., MatsudaH., SaitoS., MatsumotoK.Synthetic pyrrole–imidazole polyamide inhibits expression of the human transforming growth factor-β1 gene.J Pharmacol Exp Ther2005; 315: 571–5.
22.
MatsudaH., FukudaN., UenoT., TahiraY., AyameH., ZhangW.Development of gene silencing pyrrole–imidazole polyamide targeting the TGF-β1 promoter for treatment of progressive renal diseases.J Am Soc Nephrol2006; 17: 422–32.
23.
United States, National Institutes of Health, Institute of Laboratory Animal Resources. Guide for the Care and Use of Laboratory Animals.7th ed.Washington, DC: National Academy Press; 1996.
24.
BandoT., NaritaA., SaitoI., SugiyamaH.Molecular design of a pyrrole–imidazole hairpin polyamide for effective DNA alkylation.Chemistry2002; 8: 4781–90.
25.
YingW.Z., SandersP.W.Dietary salt modulates renal production of transforming growth factor-beta in rats.Am J Physiol1998; 274: F635–41.
26.
IshiiY., SawadaT., ShimizuA., TojimbaraT., NakajimaI., FuchinoueS.An experimental sclerosing encapsulating peritonitis model in mice.Nephrol Dial Transplant2001; 16: 1262–6.
27.
LeaskA., AbrahamD.J.TGF-β signaling and the fibrotic response.FASEB J2004; 18: 816–27.
28.
MatsudaH., FukudaN., UenoT., KatakawaM., WangX., WatanabeT.Transcriptional inhibition of progressive renal disease by gene silencing pyrrole-imidazole poly-amide targeting of the transforming growth factor-β1 promoter.Kidney Int2011; 79: 46–56.
29.
YaoE.H., FukudaN., UenoT., MatsudaH., NagaseH., MatsumotoY.A pyrrole–imidazole polyamide targeting transforming growth factor-β1 inhibits restenosis and preserves endothelialization in the injured artery.Cardiovasc Res2009; 81: 797–804.
30.
ChenM., MatsudaH., WangL., WatanabeT., KimuraM.T., IgarashiJ.Pretranscriptional regulation of TGF-β1 by PI polyamide prevents scarring and accelerates wound healing of the cornea after exposure to alkali.Mol Ther2010; 18: 519–27.
31.
DobbieJ.W.Pathogenesis of peritoneal fibrosing syndromes (sclerosing peritonitis) in peritoneal dialysis.Perit Dial Int1992; 12: 14–27.
32.
SawadaT., IshiiY., TojimbaraT., NakajimaI., FuchinoueS., TeraokaS.The ACE inhibitor, quinapril, ameliorates peritoneal fibrosis in an encapsulating peritoneal sclerosis model in mice.Pharmacol Res2002; 46: 505–10.
33.
SugaH., TeraokaS., OtaK., KomemushiS., FurutaniS., YamauchiS.Preventive effect of pirfenidone against experimental sclerosing peritonitis in rats.Exp Toxicol Pathol1995; 47: 287–91.
34.
HamaguchiA., KimS., IzumiY., ZhanY., YamanakaS., IwaoH.Contribution of extracellular signal-regulated kinase to angiotensin II–induced transforming growth factor-β1 expression in vascular smooth muscle cells.Hypertension1999; 34: 126–31.
35.
FukasawaA., AoyamaT., NagashimaT., FukudaN., UenoT., SugiyamaH.Pharmacokinetics of pyrrole–imidazole polyamide after intravenous administration in rat.Biopharm Drug Dispos2009; 30: 81–9.
36.
CombetS., MiyataT., MoulinP., PouthierD., GoffinE., DevuystO.Vascular proliferation and enhanced expression of endothelial nitric oxide synthase in human peritoneum exposed to long-term peritoneal dialysis.J Am Soc Nephrol2000; 11: 717–28.
