Preclinical Study of DNA-Recognized Peptide Compound Pyrrole-Imidazole Polyamide Targeting Human TGF-β1 Promoter for Progressive Renal Diseases in the Common Marmoset

Abstract

Pyrrole-imidazole (PI) polyamides are novel gene silencers that strongly bind the promoter region of target genes in a sequence-specific manner to inhibit gene transcription. We created a PI polyamide targeting human TGF-β1 (hTGF-β1). To develop this PI polyamide targeting hTGF-β1 (Polyamide) as a practical medicine for treating progressive renal diseases, we examined the effects of Polyamide in two common marmoset models of nephropathy. We performed lead optimization of PI polyamides that targeted hTGF-β1 by inhibiting in a dose-dependent manner the expression of TGF-β1 mRNA stimulated by PMA in marmoset fibroblasts. Marmosets were housed and fed with a 0.05% NaCl and magnesium diet and treated with cyclosporine A (CsA; 37.5 mg/kg/day, eight weeks) to establish chronic nephropathy. We treated the marmosets with nephropathy with Polyamide (1 mg/kg/week, four weeks). We also established a unilateral urethral obstruction (UUO) model to examine the effects of Polyamide (1 mg/kg/week, four times) in marmosets. Histologically, the renal medulla from CsA-treated marmosets showed cast formation and interstitial fibrosis in the renal medulla. Immunohistochemistry showed strong staining of Polyamide in the renal medulla from CsA-treated marmosets. Polyamide treatment (1 mg/kg/week, four times) reduced hTGF-β1 staining and urinary protein excretion in CsA-treated marmosets. In UUO kidneys from marmosets, Polyamide reduced the glomerular injury score and tubulointerstitial injury score. Polyamide significantly suppressed hTGF-β1 and snail mRNA expression in UUO kidneys from the marmosets. Polyamide effectively improved CsA- and UUO-associated nephropathy, indicating its potential application in the prevention of renal fibrosis in progressive renal diseases.

Contribution of TGF-β1 and Effects of Gene Silencer Pyrrole-Imidazole Polyamides Targeting TGF-β1 in Diabetic Nephropathy

“…We recently developed a PI polyamide to target human TGF-β1 and examined this PI polyamide in a primate model of cyclosporine A-induced nephropathy and unilateral urethral obstruction in the common marmoset, which has high gene homology of 90% to the human TGF-β1 gene. The human TGF-β1 PI polyamide effectively improved these nephropathies in the common marmoset [27]. We will, therefore, create diabetic mellitus in the common marmoset with STZ, and examine the effectiveness of human TGF-β1 PI polyamide as a preclinical study for the treatment of diabetic nephropathy.…”

Effects of Pyrrole-Imidazole Polyamides Targeting Human TGF-β1 on the Malignant Phenotypes of Liver Cancer Cells

“…Nevertheless, the binding of PI polyamide was reported to alter the conformation of double-stranded DNA promoter construction and impair target promoter activity [ 19 ]. We also reported that PI polyamides targeting the TGF-β1 promoter improved TGF-β1-induced fibrotic diseases such as progressive renal diseases [ 20 ], diabetic nephropathy [ 21 ], hypertrophic scar in marmoset [ 15 ], restenosis of arteries after injury [ 22 ], encapsulating peritoneal sclerosis [ 23 ], and suppressed dimethylnitrosamine-induced liver fibrosis [ 24 ]. In addition, pyridine derivative that inhibits TGF-β production improve HCV-related fibrosis and inflammation [ 25 ].…”

“…First, we did not confirm inhibition of cancer progression via TGF-β1 suppression by GB1101 in protein level. However, we previously reported the GB1101 effects on various types of diseases such as nephropathy [ 20 ], hypertrophic scar [ 15 ], restenosis of arteries after injury [ 22 ], peritoneal sclerosis [ 23 ], and liver fibrosis [ 24 ] using rat or marmoset models. Therefore, the perspective of this study is to evaluate whether GB1101 would actually be available to cancer therapy by in vivo study.…”

Rhein Improves Renal Fibrosis by Restoring Cpt1a-Mediated Fatty Acid Oxidation through SirT1/STAT3/twist1 Pathway

“…For histopathological assay, the sections were stained with hematoxylin-eosin (H&E) to observe pathological changes or Masson’s trichrome to evaluate collagen deposition. Renal pathological changes and collagen deposition were estimated by professionals using morphometric methods [ 33 ]. For IF assay of renal tissue, the sections were blocked with normal goat serum for 30 min, and then stained with primary antibodies of α-SMA, Col1A, or a mixture of Cpt1a and Twist1, respectively.…”