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| Status |
Public on Jul 08, 2021 |
| Title |
Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways. |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
A heterogeneous genetic subtype of B-cell precursor acute lymphoblastic leukemia is driven by constitutive kinase-activation, including patients with JAK2 fusions. In our study, we model the impact of a novel JAK2 fusion protein on hematopoietic development in human induced pluripotent stem cells (hiPSCs). We insert the RUNX1-JAK2 fusion into one endogenous RUNX1 allele through employing in trans paired nicking genome editing. Tagging of the fusion with a degron facilitates protein depletion using the heterobifunctional compound dTAG-13. Throughout in vitro hematopoietic differentiation, the expression of RUNX1-JAK2 is driven by endogenous RUNX1 regulatory elements at physiological levels. Functional analysis reveals that RUNX1-JAK2 knock-in cell lines yield fewer hematopoietic progenitors, due to RUNX1 haploinsufficiency. Nevertheless, these progenitors further differen-tiate toward myeloid lineages to a similar extent as wild-type cells. The expression of the RUNX1-JAK2 fusion protein only elicits subtle effects on myeloid differentiation, and is unable to transform early hematopoietic progenitors. However, phosphoprotein and transcriptome analyses reveal that RUNX1-JAK2 constitutively activates JAK-STAT signaling in differentiating hiPSCs and at the same time upregulates MYC targets—confirming the interaction between these pathways. This proof-of-principle study indicates that conditional expression of oncogenic fusion proteins in combination with hematopoietic differentiation of hiPSCs may be applicable to leukemia-relevant disease modeling.
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| Overall design |
Differential gene expression analysis using RNA-seq of three wild-type controls (WT) and six RUNX1-JAK2 clones (RJ), each either dTAG-13-treated or not.
|
| Web link |
https://doi.org/10.3390/ijms22147576
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| |
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| Contributor(s) |
Fortschegger K, Husa A, Schinnerl D, Nebral K, Strehl S |
| Citation(s) |
34299194 |
| |
| Submission date |
Oct 08, 2020 |
| Last update date |
Aug 02, 2021 |
| Contact name |
Klaus Josef Fortschegger |
| E-mail(s) |
[email protected]
|
| Organization name |
St. Anna Kinderkrebsforschung
|
| Department |
Genetics of Leukemia
|
| Lab |
Strehl
|
| Street address |
Zimmermannplatz 10
|
| City |
Vienna |
| State/province |
Austria |
| ZIP/Postal code |
1090 |
| Country |
Austria |
| |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
|
| Samples (18)
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| Relations |
| BioProject |
PRJNA668149 |
| SRA |
SRP286788 |
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