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Generación de modificadores de epigenoma de diseño de ARNm que codifican el anti-HBV

Prashika Singh1, Tafadzwa Mlambo2, Kristie Bloom1

  • 1Wits/SAMRC Antiviral Gene Therapy Research Unit, Infectious Diseases and Oncology Research Institute (IDORI), Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.

Methods in molecular biology (Clifton, N.J.)
|August 28, 2025
PubMed
Resumen
Este resumen es generado por máquina.

Las nuevas herramientas de edición de epigenoma utilizan modificadores de epigenoma de diseño (DEM) entregados a través del ARNm para apuntar y silenciar la replicación del virus de la hepatitis B crónica (VHB). Este enfoque ofrece una estrategia prometedora y no mutagénica para el tratamiento duradero del VHB.

Palabras clave:
Islas CpGLos DEMEl VHBTranscripción in vitro

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Área de la Ciencia:

  • La epigenética
  • Virología
  • Biología molecular

Sus antecedentes:

  • La infección crónica por el virus de la hepatitis B (VHB) afecta a más de 257 millones de personas en todo el mundo, causando una mortalidad significativa por enfermedad hepática.
  • Las terapias antivirales actuales tienen una capacidad limitada para eliminar el reservorio de ADN circular covalentemente cerrado (cccDNA) persistente del VHB.
  • La edición del epigenoma presenta una estrategia novedosa y no mutagénica para apuntar a la expresión génica viral.

Objetivo del estudio:

  • Desarrollar y evaluar modificadores del epigenoma de diseño (DEM) codificados por ARNm para el silenciamiento epigenético dirigido del VHB.
  • Optimizar la producción de ARNm mediante la eliminación del ARN de doble cadena inmunostimulante (ARNds).
  • Evaluar la eficacia de la represión transcripcional mediada por DEM del ADN del VHB en cultivos celulares.

Principales métodos:

  • Producción de DEM codificantes de ARNm diseñados para metilizar las islas de CpG en marcos de lectura abiertos clave de VHB (ORF).
  • Transcripción in vitro de las secuencias DEM de la plataforma pT7 ((AG)).
  • Purificación del ARNm para eliminar los contaminantes del ARNm, mejorando la expresión.
  • Transfección de células cultivadas con ARNm sintetizado y evaluación de la represión transcripcional del ADN del VHB.

Principales resultados:

  • Síntesis exitosa del ARNm que codifica los DEM diseñados para la modificación del epigenoma del VHB.
  • Mejora de la expresión de ARNm lograda mediante protocolos de eliminación de ARNm.
  • Se ha demostrado la represión transcripcional dirigida del ADN del VHB en células de cultivo mediadas por DEM.

Conclusiones:

  • Los DEM suministrados por ARNm representan una estrategia viable para el silenciamiento epigenético del VHB crónico.
  • Este enfoque minimiza los riesgos de integración genómica y los efectos fuera del objetivo asociados con otras tecnologías de edición de genes.
  • La terapia epigenética tiene potencial para obtener beneficios terapéuticos duraderos en la infección crónica por VHB.