Development of gene silencing pyrrole-imidazole polyamide targeting the TGF-beta1 promoter for treatment of progressive renal diseases.
Pyrrole-imidazole (Py-Im) polyamides are nuclease-resistant novel compounds that inhibit gene expression by binding to the minor groove of DNA. A Py-Im polyamide that targets rat TGF-beta1 was designed as a gene-silencing agent for progressive renal diseases, and the distribution and the effects of this polyamide on renal injury were examined in Dahl-salt sensitive (Dahl-S) rats. For identification of transcription factor binding elements for activation of the rat TGF-beta1 gene, recombinant TGF-beta1 reporter plasmids were transfected into HEK-293 cells, and promoter activity was measured. Py-Im polyamide was designed to the activator protein-1 binding site of the rat TGF-beta1 promoter. This Py-Im polyamide showed strong, fast, and specific binding to the target DNA in gel mobility shift and Biacore assays. Py-Im polyamide significantly inhibited TGF-beta1 promoter activity and expression of TGF-beta1 mRNA and protein in rat mesangial cells. Intravenously administered fluorescein-labeled polyamide distributed to the kidney of rats. Py-Im polyamide significantly inhibited expression of TGF-beta1 mRNA and protein in the renal cortex of Dahl-S rats and reduced the increase in urinary protein and albumin in Dahl-S rats independent of changes in blood pressure. These results indicate that Py-Im polyamide that targets TGF-beta1 will be a novel gene-silencing agent for the TGF-beta1-associated diseases, including progressive renal diseases.
Matsuda
Hiroyuki
H
Department of Internal Medicine, Nihon University School of Medicine, Ooyaguchi-kami 30-1, Itabashi-ku, Tokyo 173-8610, Japan.
Fukuda
Noboru
N
Ueno
Takahiro
T
Tahira
Yoshiko
Y
Ayame
Hirohito
H
Zhang
Wen
W
Bando
Toshikazu
T
Sugiyama
Hiroshi
H
Saito
Satoshi
S
Matsumoto
Koichi
K
Mugishima
Hideo
H
Serie
Kazuo
K
D000818:Animals; D018929:Cell Culture Techniques; D020868:Gene Silencing; D007093:Imidazoles / Q000494:pharmacology*; D007674:Kidney Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D050527:Mesangial Cells / Q000187:drug effects* / Q000378:metabolism; D009757:Nylons / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000502:physiology; D011758:Pyrroles / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020303:Rats, Inbred Dahl; D017208:Rats, Wistar; D018808:Transcription Factor AP-1 / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*; D053773:Transforming Growth Factor beta1
D007093:Imidazoles; D009757:Nylons; D011758:Pyrroles; D012333:RNA, Messenger; C508962:Tgfb1 protein, rat; D018808:Transcription Factor AP-1; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; C029899:imidazole