Chromosome Transplantation: Correction of the Chronic Granulomatous Disease Defect in Mouse Induced Pluripotent Stem Cells

Alessandra Castelli, Lucia Susani, Ciro Menale, Sharon Muggeo, Elena Caldana, Dario Strina, Barbara Cassani, Camilla Recordati, Eugenio Scanziani, Francesca Ficara, Anna Villa, Paolo Vezzoni, Marianna Paulis

Research output: Contribution to journalArticle

Abstract

In spite of the progress in gene editing achieved in recent years, a subset of genetic diseases involving structural chromosome abnormalities, including aneuploidies, large deletions and complex rearrangements, cannot be treated with conventional gene therapy approaches. We have previously devised a strategy, dubbed chromosome transplantation (CT), to replace an endogenous mutated chromosome with an exogenous normal one. To establish a proof of principle for our approach, we chose as disease model the chronic granulomatous disease (CGD), an X-linked severe immunodeficiency due to abnormalities in CYBB (GP91) gene, including large genomic deletions. We corrected the gene defect by CT in induced pluripotent stem cells (iPSCs) from a CGD male mouse model. The Hprt gene of the endogenous X chromosome was inactivated by CRISPR/Cas9 technology thus allowing the exploitation of the hypoxanthine–aminopterin–thymidine selection system to introduce a normal donor X chromosome by microcell-mediated chromosome transfer. X-transplanted clones were obtained, and diploid XY clones which spontaneously lost the endogenous X chromosome were isolated. These cells were differentiated toward the myeloid lineage, and functional granulocytes producing GP91 protein were obtained. We propose the CT approach to correct iPSCs from patients affected by other X-linked diseases with large deletions, whose treatment is still unsatisfactory. Stem Cells 2019;37:876–887.

Original languageEnglish
Pages (from-to)876-887
Number of pages12
JournalStem Cells
Volume37
Issue number7
DOIs
Publication statusPublished - Jul 2019

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Keywords

  • Chronic granulomatous disease
  • CRISPR/Cas system
  • Genetic therapy
  • Induced pluripotent stem cells
  • X-linked combined immunodeficiency diseases

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Cite this

Castelli, A., Susani, L., Menale, C., Muggeo, S., Caldana, E., Strina, D., Cassani, B., Recordati, C., Scanziani, E., Ficara, F., Villa, A., Vezzoni, P., & Paulis, M. (2019). Chromosome Transplantation: Correction of the Chronic Granulomatous Disease Defect in Mouse Induced Pluripotent Stem Cells. Stem Cells, 37(7), 876-887. https://doi.org/10.1002/stem.3006