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1.
Sci Rep ; 6: 20019, 2016 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-26883577

RESUMO

In Friedreich's ataxia (FRDA) patients, diminished frataxin (FXN) in sensory neurons is thought to yield the predominant pathology associated with disease. In this study, we demonstrate successful usage of RNA transcript therapy (RTT) as an exogenous human FXN supplementation strategy in vitro and in vivo, specifically to dorsal root ganglia (DRG). Initially, 293 T cells were transfected with codon optimized human FXN mRNA, which was translated to yield FXN protein. Importantly, FXN was rapidly processed into the mature functional form of FXN (mFXN). Next, FXN mRNA, in the form of lipid nanoparticles (LNPs), was administered intravenously in adult mice. Examination of liver homogenates demonstrated efficient FXN LNP uptake in hepatocytes and revealed that the mitochondrial maturation machinery had efficiently processed all FXN protein to mFXN in ~24 h in vivo. Remarkably, greater than 50% mFXN protein derived from LNPs was detected seven days after intravenous administration of FXN LNPs, suggesting that the half-life of mFXN in vivo exceeds one week. Moreover, when FXN LNPs were delivered by intrathecal administration, we detected recombinant human FXN protein in DRG. These observations provide the first demonstration that RTT can be used for the delivery of therapeutic mRNA to DRG.


Assuntos
Ataxia de Friedreich/genética , Gânglios Espinais/metabolismo , Proteínas de Ligação ao Ferro/genética , Lipídeos , Nanopartículas , RNA Mensageiro , Animais , Modelos Animais de Doenças , Feminino , Ataxia de Friedreich/diagnóstico , Ataxia de Friedreich/metabolismo , Ataxia de Friedreich/terapia , Expressão Gênica , Genes Reporter , Humanos , Injeções Espinhais , Proteínas de Ligação ao Ferro/metabolismo , Lipídeos/química , Fígado/metabolismo , Medições Luminescentes , Camundongos , Imagem Molecular , Nanopartículas/administração & dosagem , Nanopartículas/química , Biossíntese de Proteínas , RNA Mensageiro/administração & dosagem , RNA Mensageiro/química , Transdução de Sinais , Transfecção , Frataxina
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