Preclinical Study of DNA-Recognized Peptide Compound Pyrrole-Imidazole Polyamide Targeting Human TGF-β1 Promoter for Progressive Renal Diseases in the Common Marmoset
Preclinical Study of DNA-Recognized Peptide Compound Pyrrole-Imidazole Polyamide Targeting Human TGF-β1 Promoter for Progressive Renal Diseases in the Common Marmoset
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DOI：
10.3390/molecules24173178
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发表时间：
2019-09-01
期刊：
影响因子：
4.6
通讯作者：
Abe, Masanori
中科院分区：
文献类型：
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作者：
Otsuki, Masari;Fukuda, Noboru;Abe, Masanori
吡咯-咪唑（PI）聚酰胺是一种新型的基因沉默剂，它能以序列特异性的方式与靶基因的启动子区结合，从而抑制基因的转录。我们创建了靶向人TGF-β 1（hTGF-β 1）的PI聚酰胺。为了开发这种靶向hTGF-β 1的PI聚酰胺（聚酰胺）作为治疗进行性肾脏疾病的实用药物，我们在两种常见的绒猴肾病模型中检查了聚酰胺的作用。我们通过以剂量依赖性方式抑制由PMA刺激的绒猴成纤维细胞中TGF-β 1 mRNA的表达，对靶向TGF-β 1的PI聚酰胺进行了先导优化。饲养绒猴，用0.05%NaCl和镁饲料喂养，并用环孢霉素A（CsA; 37.5 mg/kg/天，8周）处理，以建立慢性肾病。我们用聚酰胺（1 mg/kg/周，4周）治疗患有肾病的绒猴。我们还建立了单侧尿道梗阻（UUO）模型，以检查聚酰胺（1 mg/kg/周，4次）对绒猴的影响。组织学上，CsA处理过的绒猴肾髓质表现为管型形成和肾髓质间质纤维化。免疫组织化学显示，在CsA处理的绒猴肾髓质中的聚酰胺强染色。聚酰胺处理（1 mg/kg/周，4次）减少了CsA处理的绒猴的hTGF-β 1染色和尿蛋白排泄。在来自绒猴的UUO肾脏中，聚酰胺降低了肾小球损伤评分和肾小管间质损伤评分。聚酰胺显着抑制hTGF-β 1和snail mRNA的表达在UUO从绒猴肾脏。聚酰胺可有效改善CsA和UUO相关性肾病，表明其在预防进行性肾病肾纤维化中的潜在应用。
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-beta 1 (hTGF-beta 1). To develop this PI polyamide targeting hTGF-beta 1 (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-beta 1 by inhibiting in a dose-dependent manner the expression of TGF-beta 1 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-beta 1 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-beta 1 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.