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Un sistema de pseudovirus permite el escaneo mutativo profundo del pico completo de SARS-CoV-2
Bernadeta Dadonaite1, Katharine H D Crawford2, Caelan E Radford3
1Basic Sciences Division and Computational Biology Program, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA.
Cell
|March 3, 2023
Resumen
Una nueva plataforma de exploración de mutaciones profundas mide con precisión cómo miles de mutaciones de la proteína del pico del SARS-CoV-2 afectan la neutralización de anticuerpos y la infección viral. Este método seguro y de alto rendimiento ayuda a comprender la evolución viral y el escape de anticuerpos.
Área de la Ciencia:
- Virología
- Biología molecular
- Inmunología
Sus antecedentes:
- Comprender la evolución del SARS-CoV-2 requiere analizar las mutaciones en la proteína del pico.
- El impacto de numerosas mutaciones en la neutralización de anticuerpos y la función viral sigue siendo un desafío.
Objetivo del estudio:
- Desarrollar y aplicar una plataforma de alto rendimiento para cuantificar los efectos de las mutaciones de la proteína del pico del SARS-CoV-2.
- Mapear las mutaciones de escape de anticuerpos y evaluar su impacto en la infección viral.
Principales métodos:
- Utilizó lentivirus pseudotipados no replicativos para el escaneo mutativo profundo.
- Generó grandes bibliotecas de mutaciones para las proteínas de punta Omicron BA.1 y Delta (∼7,000 mutaciones, ∼135,000 combinaciones).
- Neutralización cuantificada de anticuerpos y infección por pseudovirus a través de combinaciones de mutaciones.
Principales resultados:
- Se mapearon con éxito miles de mutaciones punzantes del SARS-CoV-2 que afectan la neutralización de anticuerpos.
- Se identificaron mutaciones de escape dirigidas al dominio de unión al receptor, al dominio N-terminal y a la subunidad S2.
- Demostró un método seguro de alto rendimiento para evaluar los efectos de la mutación en la fuga de anticuerpos y la infección.
Conclusiones:
- Estableció una plataforma robusta para estudiar la evolución de la proteína del pico viral y el escape de anticuerpos.
- La plataforma permite la medición precisa de los efectos de las combinaciones de mutaciones ∼10^5.
- Este enfoque es adaptable para analizar las proteínas de entrada de otros virus.
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