Mucopolysaccharidosis type IVB (MPSIVB) is a lysosomal storage disorder caused by (3-galactosidase ((3 (3-GAL) deficiency fi ciency characterized by severe skeletal and neurological alterations without approved treatments. To develop hematopoietic stem progenitor cell (HSPC) gene therapy (GT) for MPSIVB, we designed lentiviral vectors (LVs) encoding human (3-GAL to achieve supraphysiological release of the therapeutic enzyme in human HSPCs and metabolic correction of diseased cells. Transduced HSPCs displayed proper colony formation, proliferation, and differentiation capacity, but their progeny failed to release the enzyme at supraphysiological levels. Therefore, we tested alternative LVs to overexpress an enhanced (3-GAL deriving from murine (LV-enhGLB1) and human selectively mutated GLB1 sequences (LV-mutGLB1). Only human HSPCs transduced with LVenhGLB1 overexpressed (3-GAL in vitro and in vivo without evidence of overexpression-related toxicity. Their hematopoietic progeny efficiently fi ciently released (3-GAL, allowing the cross-correction of defective cells, including skeletal cells. We found that the low levels of human GLB1 mRNA in human hematopoietic cells and the improved stability of the enhanced (3-GAL contribute to the increased efficacy fi cacy of LV-enhGLB1. Importantly, the enhanced (3-GAL enzyme showed physiological lysosomal trafficking fi cking in human cells and was not associated with increased immunogenicity in vitro. These results support the use of LVenhGLB1 for further HSPC-GT development and future clinical translation to treat MPSIVB multisystem disease.

A GLB1 transgene with enhanced therapeutic potential for the preclinical development of ex-vivo gene therapy to treat mucopolysaccharidosis type IVB / Crippa, Stefania; Alberti, Gaia; Passerini, Laura; Savoia, Evelyn Oliva; Mancino, Marilena; De Ponti, Giada; Santi, Ludovica; Berti, Margherita; Testa, Marialuisa; Hernandez, Raisa Jofra; Quaranta, Pamela; Ceriotti, Selene; Visigalli, Ilaria; Morrone, Amelia; Paoli, Antonella; Forni, Claudia; Scala, Serena; Degano, Massimo; Staiano, Leopoldo; Gregori, Silvia; Aiuti, Alessandro; Bernardo, Maria Ester. - In: MOLECULAR THERAPY. METHODS & CLINICAL DEVELOPMENT. - ISSN 2329-0501. - ELETTRONICO. - Volume 32(3); 2024 Sep 12:(2024), pp. eCollection 2024 Sep 12.0-eCollection 2024 Sep 12.0. [10.1016/j.omtm.2024.101313]

A GLB1 transgene with enhanced therapeutic potential for the preclinical development of ex-vivo gene therapy to treat mucopolysaccharidosis type IVB

Morrone, Amelia;Forni, Claudia;
2024

Abstract

Mucopolysaccharidosis type IVB (MPSIVB) is a lysosomal storage disorder caused by (3-galactosidase ((3 (3-GAL) deficiency fi ciency characterized by severe skeletal and neurological alterations without approved treatments. To develop hematopoietic stem progenitor cell (HSPC) gene therapy (GT) for MPSIVB, we designed lentiviral vectors (LVs) encoding human (3-GAL to achieve supraphysiological release of the therapeutic enzyme in human HSPCs and metabolic correction of diseased cells. Transduced HSPCs displayed proper colony formation, proliferation, and differentiation capacity, but their progeny failed to release the enzyme at supraphysiological levels. Therefore, we tested alternative LVs to overexpress an enhanced (3-GAL deriving from murine (LV-enhGLB1) and human selectively mutated GLB1 sequences (LV-mutGLB1). Only human HSPCs transduced with LVenhGLB1 overexpressed (3-GAL in vitro and in vivo without evidence of overexpression-related toxicity. Their hematopoietic progeny efficiently fi ciently released (3-GAL, allowing the cross-correction of defective cells, including skeletal cells. We found that the low levels of human GLB1 mRNA in human hematopoietic cells and the improved stability of the enhanced (3-GAL contribute to the increased efficacy fi cacy of LV-enhGLB1. Importantly, the enhanced (3-GAL enzyme showed physiological lysosomal trafficking fi cking in human cells and was not associated with increased immunogenicity in vitro. These results support the use of LVenhGLB1 for further HSPC-GT development and future clinical translation to treat MPSIVB multisystem disease.
2024
Volume 32(3); 2024 Sep 12
0
0
Crippa, Stefania; Alberti, Gaia; Passerini, Laura; Savoia, Evelyn Oliva; Mancino, Marilena; De Ponti, Giada; Santi, Ludovica; Berti, Margherita; Testa...espandi
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in FLORE sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1388452
Citazioni
  • ???jsp.display-item.citation.pmc??? 0
  • Scopus ND
  • ???jsp.display-item.citation.isi??? 0
social impact