Identification of hematopoietic stem cell-specific miRNAs enables gene therapy of globoid cell leukodystrophy

Bernhard Gentner, Ilaria Visigalli, Hidefumi Hiramatsu, Eric Lechman, Silvia Ungari, Alice Giustacchini, Giulia Schira, Mario Amendola, Angelo Quattrini, Sabata Martino, Aldo Orlacchio, John E. Dick, Alessandra Biffi, Luigi Naldini

Research output: Contribution to journalArticle

Abstract

Globoid cell leukodystrophy (GLD; also known as Krabbe disease) is an invariably fatal lysosomal storage disorder caused by mutations in the galactocerebrosidase (GALC) gene. Hematopoietic stem cell (HSC)-based gene therapy is being explored for GLD; however, we found that forced GALC expression was toxic to HSCs and early progenitors, highlighting the need for improved regulation of vector expression. We used a genetic reporter strategy based on lentiviral vectors to detect microRNA activity in hematopoietic cells at single-cell resolution. We report that miR-126 and miR-130a were expressed in HSCs and early progenitors from both mice and humans, but not in differentiated progeny. Moreover, repopulating HSCs could be purified solely on the basis of miRNA expression, providing a new method relevant for human HSC isolation. By incorporating miR-126 target sequences into a GALC-expressing vector, we suppressed GALC expression in HSCs while maintaining robust expression in mature hematopoietic cells. This approach protected HSCs from GALC toxicity and allowed successful treatment of a mouse GLD model, providing a rationale to explore HSC-based gene therapy for GLD.

Original languageEnglish
Article number58ra84
JournalScience Translational Medicine
Volume2
Issue number58
DOIs
Publication statusPublished - Nov 17 2010

ASJC Scopus subject areas

  • Medicine(all)

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    Gentner, B., Visigalli, I., Hiramatsu, H., Lechman, E., Ungari, S., Giustacchini, A., Schira, G., Amendola, M., Quattrini, A., Martino, S., Orlacchio, A., Dick, J. E., Biffi, A., & Naldini, L. (2010). Identification of hematopoietic stem cell-specific miRNAs enables gene therapy of globoid cell leukodystrophy. Science Translational Medicine, 2(58), [58ra84]. https://doi.org/10.1126/scitranslmed.3001522