New models for the most frequent Mucolipidosis II-causing mutation using iPSCs and zebrafish: a crucial step towards the development of new therapies

Publicações

Establishment of a Human iPSC Line from Mucolipidosis Type II That Expresses the Key Markers of the Disease

Publication . Moutinho, Maria Eduarda; Gonçalves, Mariana; Duarte, Ana J.; Encarnação, Marisa; Coutinho, Maria Francisca; Matos, Liliana; Santos, Juliana I.; Ribeiro, Diogo; Amaral, Olga; Gaspar, Paulo; Alves, Sandra; Moreira, Luciana V.

Mucolipidosis type II (ML II) is a rare and fatal disease of acid hydrolase trafficking. It is caused by pathogenic variants in the GNPTAB gene, leading to the absence of GlcNAc-1-phosphotransferase activity, an enzyme that catalyzes the first step in the formation of the mannose 6-phosphate (M6P) tag, essential for the trafficking of most lysosomal hydrolases. Without M6P, these do not reach the lysosome, which accumulates undegraded substrates. The lack of samples and adequate disease models limits the investigation into the pathophysiological mechanisms of the disease and potential therapies. Here, we report the generation and characterization of an ML II induced pluripotent stem cell (iPSC) line carrying the most frequent ML II pathogenic variant [NM_024312.5(GNPTAB):c.3503_3504del (p.Leu1168fs)]. Skin fibroblasts were successfully reprogrammed into iPSCs that express pluripotency markers, maintain a normal karyotype, and can differentiate into the three germ layers. Furthermore, ML II iPSCs showed a phenotype comparable to that of the somatic cells that originated them in terms of key ML II hallmarks: lower enzymatic activity of M6P-dependent hydrolases inside the cells but higher in conditioned media, and no differences in an M6P-independent hydrolase and accumulation of free cholesterol. Thus, ML II iPSCs constitute a novel model for ML II disease, with the inherent iPSC potential to become a valuable model for future studies on the pathogenic mechanisms and testing potential therapeutic approaches.

2025-04-19Artigo científico

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Palavras-chave

CRISPR/Cas9,lysosomal storage disorder,in vitro model,in vivo model, Medical and health sciences

Entidade financiadora

Fundação para a Ciência e a Tecnologia, I.P.
Fundação para a Ciência e a Tecnologia, I.P.

Programa de financiamento

Concurso de Projetos de I&D em Todos os Domínios Científicos - 2022 - PEX
Concurso de Projetos de I&D em Todos os Domínios Científicos - 2022

Número da atribuição

2022.03836.PTDC