The Leukemic Isocitrate Dehydrogenase (IDH) 1/2 Mutations Impair Myeloid and Erythroid Cell Differentiation of Primary Human Hematopoietic Stem and Progenitor Cells (HSPCs)
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Human CD34+ Hematopoietic Stem and Progenitor Cells from Healthy Donors
2.2. Sequences and Expression Vector
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- Homo sapiens isocitrate dehydrogenase (NADP(+)) 1, cytosolic (IDH1), transcript variant 1, mRNA (NCBI reference sequence: NM_005896.3).
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- Homo sapiens isocitrate dehydrogenase (NADP(+)) 2, mitochondrial (IDH2), transcript variant 1, mRNA (NCBI reference sequence: NM_002168.3).
- -
- Mutated sequences were virtually designed according to the most common nucleotide change in cytogenetically normal–AML [17]:
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- IDH1-R132H (nucleotide change c.395G>A).
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- IDH2-R140Q (nucleotide change c.419G>A).
2.3. Lentivirus Production
2.4. hCD34+ Lentiviral Infection
2.5. hCD34+ Cell-Staining and Sorting Strategies
2.6. hCD34+ Cells, Colony-Forming Unit Assays
2.7. CD34+ Cells from a IDH2-Mutated AML Patient
2.8. Next-Generation Targeted-DNA Sequencing (NGS)
2.9. Statistical Analysis
3. Results
3.1. Either the IDH1-R132H or IDH2-R140Q Mutation Blocks CFU Ability of Human CD34+ HSPC
3.2. Enasidenib (AG-221) Treatment Induces a Progressive Improvement of CFU Ability of Primary CD34+ Cells in a Patient with IDH2-Mutated AML
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Pierangeli, S.; Donnini, S.; Ciaurro, V.; Milano, F.; Cardinali, V.; Sciabolacci, S.; Cimino, G.; Gionfriddo, I.; Ranieri, R.; Cipriani, S.; et al. The Leukemic Isocitrate Dehydrogenase (IDH) 1/2 Mutations Impair Myeloid and Erythroid Cell Differentiation of Primary Human Hematopoietic Stem and Progenitor Cells (HSPCs). Cancers 2024, 16, 2675. https://doi.org/10.3390/cancers16152675
Pierangeli S, Donnini S, Ciaurro V, Milano F, Cardinali V, Sciabolacci S, Cimino G, Gionfriddo I, Ranieri R, Cipriani S, et al. The Leukemic Isocitrate Dehydrogenase (IDH) 1/2 Mutations Impair Myeloid and Erythroid Cell Differentiation of Primary Human Hematopoietic Stem and Progenitor Cells (HSPCs). Cancers. 2024; 16(15):2675. https://doi.org/10.3390/cancers16152675
Chicago/Turabian StylePierangeli, Sara, Serena Donnini, Valerio Ciaurro, Francesca Milano, Valeria Cardinali, Sofia Sciabolacci, Gaetano Cimino, Ilaria Gionfriddo, Roberta Ranieri, Sabrina Cipriani, and et al. 2024. "The Leukemic Isocitrate Dehydrogenase (IDH) 1/2 Mutations Impair Myeloid and Erythroid Cell Differentiation of Primary Human Hematopoietic Stem and Progenitor Cells (HSPCs)" Cancers 16, no. 15: 2675. https://doi.org/10.3390/cancers16152675