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Keratinocyte cell lines derived from severe generalized recessive E pidermolysis B ullosa patients carrying a highly recurrent COL 7A1 homozygous mutation: models to assess cell and gene therapies in vitro and in vivo

2013; Wiley; Volume: 22; Issue: 9 Linguagem: Inglês

10.1111/exd.12203

1600-0625

Cristina Chamorro, David Almarza, Blanca Duarte, Sara Llames, Rodolfo Murillas, Marta García, Juan C. Cigudosa, Luis Espinosa‐Hevia, M.J. Escámez, Ángeles Mencía, Álvaro Meana, Ramón García‐Escudero, Rosa Moro, Claudio J. Conti, Marcela Del Río, Fernando Larcher,

Silk-based biomaterials and applications

Recessive dystrophic epidermolysis bullosa (RDEB) is caused by deficiency of type VII collagen due to COL7A1 mutations such as c.6527insC, recurrently found in the Spanish RDEB population. Assessment of clonal correction-based therapeutic approaches for RDEB requires large expansions of cells, exceeding the replication capacity of human primary keratinocytes. Thus, immortalized RDEB cells with enhanced proliferative abilities would be valuable. Using either the SV40 large T antigen or papillomavirus HPV16-derived E6-E7 proteins, we immortalized and cloned RDEB keratinocytes carrying the c.6527insC mutation. Clones exhibited high proliferative and colony-forming features. Cytogenetic analysis revealed important differences between T antigen-driven and E6-E7-driven immortalization. Immortalized cells responded to differentiation stimuli and were competent for epidermal regeneration and recapitulation of the blistering RDEB phenotype in vivo. These features make these cell lines useful to test novel therapeutic approaches including those aimed at editing mutant COL7A1.