Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways
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 (h...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7576 |
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| author | Klaus Fortschegger Anna-Maria Husa Dagmar Schinnerl Karin Nebral Sabine Strehl |
| author_facet | Klaus Fortschegger Anna-Maria Husa Dagmar Schinnerl Karin Nebral Sabine Strehl |
| author_sort | Klaus Fortschegger |
| collection | DOAJ |
| description | 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 <i>RUNX1-JAK2</i> fusion into one endogenous <i>RUNX1</i> 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 <i>RUNX1</i> regulatory elements at physiological levels. Functional analysis reveals that <i>RUNX1-JAK2</i> knock-in cell lines yield fewer hematopoietic progenitors, due to <i>RUNX1</i> haploinsufficiency. Nevertheless, these progenitors further differentiate 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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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:37:35Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c005a61b4f5b4bba95ac77e183e90b6f2023-11-22T04:00:46ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214757610.3390/ijms22147576Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC PathwaysKlaus Fortschegger0Anna-Maria Husa1Dagmar Schinnerl2Karin Nebral3Sabine Strehl4St. Anna Children’s Cancer Research Institute (CCRI), 1090 Vienna, AustriaSt. Anna Children’s Cancer Research Institute (CCRI), 1090 Vienna, AustriaSt. Anna Children’s Cancer Research Institute (CCRI), 1090 Vienna, AustriaSt. Anna Children’s Cancer Research Institute (CCRI), 1090 Vienna, AustriaSt. Anna Children’s Cancer Research Institute (CCRI), 1090 Vienna, AustriaA 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 <i>RUNX1-JAK2</i> fusion into one endogenous <i>RUNX1</i> 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 <i>RUNX1</i> regulatory elements at physiological levels. Functional analysis reveals that <i>RUNX1-JAK2</i> knock-in cell lines yield fewer hematopoietic progenitors, due to <i>RUNX1</i> haploinsufficiency. Nevertheless, these progenitors further differentiate 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.https://www.mdpi.com/1422-0067/22/14/7576leukemiaoncogenic fusionCRISPR/Cas9hematopoiesishiPSCJAK-STAT signaling |
| spellingShingle | Klaus Fortschegger Anna-Maria Husa Dagmar Schinnerl Karin Nebral Sabine Strehl Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways International Journal of Molecular Sciences leukemia oncogenic fusion CRISPR/Cas9 hematopoiesis hiPSC JAK-STAT signaling |
| title | Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways |
| title_full | Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways |
| title_fullStr | Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways |
| title_full_unstemmed | Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways |
| title_short | Expression of RUNX1-JAK2 in Human Induced Pluripotent Stem Cell-Derived Hematopoietic Cells Activates the JAK-STAT and MYC Pathways |
| title_sort | expression of runx1 jak2 in human induced pluripotent stem cell derived hematopoietic cells activates the jak stat and myc pathways |
| topic | leukemia oncogenic fusion CRISPR/Cas9 hematopoiesis hiPSC JAK-STAT signaling |
| url | https://www.mdpi.com/1422-0067/22/14/7576 |
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