Therapeutic Effects of Long-Term Administration of Tranilast in an Animal Model for the Treatment of Fibroids

Abstract

Tranilast (N-3, 4-dimethoxycinnamoyl anthranilic acid) is an orally administered drug with antiallergic properties and approved in Japan and the Republic of Korea for the treatment of asthma and hypertrophic scars. Previous in vitro studies indicated that tranilast reduced fibroid growth through its inhibitory effects on cell proliferation and induction of apoptosis. The objective of this study was to determine the efficacy of tranilast for treatment of human-derived fibroids in a mouse model. SCID mice (ovariectomized, supplemented with estrogen and progesterone) were implanted with fibroid explants and treated for two months with tranilast (50 m/kg/daily) or the vehicle. After sacrifice, xenografts were excised and analyzed. Tranilast was well tolerated without adverse side effects. There was a 37% reduction in tumor weight along with a significant decrease in staining for Ki67, CCND1, and E2F1; a significant increase in nuclear staining for cleaved caspase 3; and reduced staining for TGF-β3 and Masson’s trichrome in the tranilast treated mice. There was a significant inhibition of mRNA and protein expression of fibronectin, COL3A1, CCND1, E2F1, and TGF-β3 in the xenografts from the tranilast-treated mice. These promising therapeutic effects of tranilast warrant additional animal studies and human clinical trials to evaluate its efficacy for treatment of fibroids.

Targeting the long non-coding RNA MIAT for the treatment of fibroids in an animal model

“…Alternative therapies that do not inhibit estrogen production causing hypoestrogenic side effects are urgently needed for treatment of fibroids. We have previously provided preclinical in vivo data for two therapies in a mouse model, including treatment with the anti-inflammatory drug tranilast [ 14 ] and more recently using a drug that inhibits the activity of the tryptophan catabolizing enzyme TDO2 [ 15 ] which is markedly overexpressed in fibroids [ 43 , 44 ]. Because of their localized presence and slow growth treatment of fibroids by gene therapy approaches have attracted some attention with some promising results.…”

“…The plasma chemistry was analyzed by the VetScan VS2 chemistry analyzer (Abaxis, Union City, CA, U.S.A.) (Comprehensive Diagnostic Profile Rotor (#500-1038, Abaxis, Union City, CA, U.S.A.) according to the manufacturer’s protocol as previously described [ 14 , 15 ].…”

“…Five-micron sections were cut and mounted on glass slides. The Masson’s trichrome stain (HT15-1KT, Sigma-Aldrich) and the immunohistochemistry were performed as previously described [ 14 , 15 ]. The primary antibodies used in this study are rabbit anti-Ki67 (dilution 1:250, 27309-1-AP, Proteintech Group, Inc, Rosemont, IL, U.S.A.), rabbit anti-cleaved caspase-3 (dilution 1:50, #9664, Cell Signaling Technology, Danvers, MA, U.S.A.), rabbit anti-COL3A1 (dilution 1:1000, 22734-1-AP, Proteintech Group, Inc, Rosemont, IL, U.S.A.), and mouse anti-E2F1 (dilution 1:50, sc-251, Santa Cruz Biotechnology, Inc., Dallas, TX, U.S.A.).…”

“…Based on these findings we hypothesized that downregulation of MIAT would result in up-regulation of miR-29 which in turn would result in down-regulation of target genes for this key miRNA including genes regulating ECM composition such as collagen and TGF-β3 and genes that regulate the cell cycle such as CDK2. We tested this hypothesis in a mouse model for fibroids which we have previously characterized [ 14 , 15 ].…”

The Effect of Race/Ethnicity and MED12 Mutation on the Expression of Long Non-Coding RNAs in Uterine Leiomyoma and Myometrium

“…Total RNA was extracted from leiomyoma and matched myometrium (n = 19) using TRIzol (Thermo Fisher Scientific Inc., Waltham, MA, USA). RNA concentration and integrity was determined using a Nanodrop 2000c spectrophotometer (Thermo Scientific, Wilmington, DE, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) as previously described [29,96]. Samples with RNA integrity numbers (RIN) greater than or equal to 9 were used for library preparation.…”