TY - JOUR
T1 - Chromosome Transplantation
T2 - Correction of the Chronic Granulomatous Disease Defect in Mouse Induced Pluripotent Stem Cells
AU - Castelli, Alessandra
AU - Susani, Lucia
AU - Menale, Ciro
AU - Muggeo, Sharon
AU - Caldana, Elena
AU - Strina, Dario
AU - Cassani, Barbara
AU - Recordati, Camilla
AU - Scanziani, Eugenio
AU - Ficara, Francesca
AU - Villa, Anna
AU - Vezzoni, Paolo
AU - Paulis, Marianna
PY - 2019/7
Y1 - 2019/7
N2 - 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.
AB - 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.
KW - Chronic granulomatous disease
KW - CRISPR/Cas system
KW - Genetic therapy
KW - Induced pluripotent stem cells
KW - X-linked combined immunodeficiency diseases
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U2 - 10.1002/stem.3006
DO - 10.1002/stem.3006
M3 - Article
C2 - 30895693
AN - SCOPUS:85063947273
VL - 37
SP - 876
EP - 887
JO - Stem Cells
JF - Stem Cells
SN - 1066-5099
IS - 7
ER -