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| Status |
Public on May 29, 2024 |
| Title |
Mis-splicing of mitotic regulators sensitizes SF3B1-mutated human HSCs to CHK1 inhibition [scRNA-seq] |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Splicing factor SF3B1 mutations are frequent somatic lesions in myeloid neoplasms that transform hematopoietic stem cells (HSCs) by inducing mis-splicing of target genes. However, the molecular and functional consequences of SF3B1 mutations in human HSCs remain unclear. Here, we identify the mis-splicing program in human HSCs as a targetable vulnerability by precise gene editing of SF3B1 K700E mutations in primary CD34+ cells. Mutant SF3B1 induced pervasive mis-splicing and reduced expression of genes regulating mitosis in single cell transcriptomics, critically BUBR1 and CDC27, leading to altered differentiation and delayed G2/M progression. Mutant SF3B1 mis-splicing or reduced expression of BUBR1 and CDC27 was sufficient to delay G2/M transit, leading to activation of CHK1, sensitizing cells to CHK1 inhibition. Clinical CHK1 inhibitor prexasertib selectively targeted SF3B1-mutant HSCs and abrogated engraftment in vivo. These findings identify mis-splicing of mitotic regulators in SF3B1-mutant HSCs as a targetable vulnerability engaged by pharmacological CHK1 inhibition.
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| Overall design |
Single cell RNA sequencing was performed on SF3B1 K700E mutant HSPCs and AAVS1 gene edited controls. Gene edited BFP+CD34+ HSPCs were flow-sorted 72 hours after gene editing.
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| Contributor(s) |
Sarchi M, Clough CA, Crosse EI, Kim J, Baquero Galvis LD, Aydinyan N, Wellington R, Gallì A, Creamer JP, Stewart S, Bradley RK, Malcovati L, Doulatov S |
| Citation(s) |
38856693 |
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| Submission date |
Apr 05, 2024 |
| Last update date |
Aug 29, 2024 |
| Contact name |
Martina Sarchi |
| E-mail(s) |
martina.sarchi@unipv.it, doulatov@uw.edu
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| Organization name |
University of Pavia
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| Department |
Dept. of Molecular Medicine
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| Street address |
Via Ferrata 9
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| City |
Pavia |
| ZIP/Postal code |
27100 |
| Country |
Italy |
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| Platforms (1) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA1096664 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE263300_RAW.tar |
980.0 Mb |
(http)(custom) |
TAR (of H5AD) |
| GSE263300_filtered_ad.h5ad |
876.9 Mb |
(ftp)(http) |
H5AD |
SRA Run Selector |
| Raw data are available in SRA |
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