Retinoid X receptor alpha is a spatiotemporally-specific therapeutic target for doxorubicin-induced cardiomyopathy in adult zebrafish

Abstract

While the genetic suppressor screen is efficient in suggesting therapeutic genes, this strategy has yet to be successful for cardiomyopathies in vertebrates. To develop such a strategy, we recently established a mutagenesis screen platform in zebrafish for systematic discovery of genetic modifiers of doxorubicin-induced cardiomyopathy (DIC). Here, we further revealed both molecular and cellular insights of the first salutary modifier emerged from the screen, i.e. gene-breaking transposon (GBT) 0419 that affects the retinoid X receptor alpha a (rxraa) gene.First, by rescuing the mutation in tissue-specific manner with multiple Cre-loxP systems, we demonstrated that the endothelial, but not myocardial or epicardial, function of rxraa is primary to this cardioprotective effects. Next, we showed that the rxraa-associated salutary effects on DIC were conferred partially by the activation of retinoid acid (RA) signaling. Finally, we identified isotretinoin and bexarotene, 2 US Food and Drug Administration-approved RXRA agonists that are effective in treating adult zebrafish DIC when administered during the early, but not the late, phase of DIC progression. Collectively, we provided the first in vivo genetic evidence in supporting RXRA as the therapeutic target for DIC, and uncovered a previously unrecognized spatiotemporally-restricted mechanism for this gene-based therapeutic strategy. Our study also justified that searching salutary modifiers via zebrafish mutagenesis screen can be effective in discovering new therapeutic targets for cardiomyopathies.

Disruption of MAP7D1 Gene Function Increases the Risk of Doxorubicin-Induced Cardiomyopathy and Heart Failure

“…Thus, a major novelty of this study is to combine the power of zebrafish genetics and a human GWAS for both identification and functional elucidation of MAP7D1 as a clinically relevant susceptibility gene to DIC and heart failure. The feasibility of this approach was further supported by recent studies that reported RXRA as a genetic factor associated with anthracycline-induced cardiotoxicity (AIC) [13,15]. This approach, we believe, can be readily applied to study other AIC genes as well.…”

“…Echocardiography. Echocardiography measurement was carried out using the Vevo 3100 high-frequency imaging system equipped with a 50 MHz (MX700) linear array transducer (FUJIFILM VisualSonics Inc.) accordingly [15]. Briefly, adult zebrafish at appropriate stages were anesthetized in tricaine (0.016%) for 5 minutes, placed ventral side up into a sponge.…”

“…Swimming Tunnel Assay. The swimming tunnel protocol was conducted using a swim tunnel respirometer (Mini Swim 170, Loligo Systems, Denmark) according to the previous reports [15]. Both GBT239 mutant and WT fish after doxorubicin injection were fasted for 24 h prior to swimming capacity measurement.…”