Nup98-Kdm5a, induced via CRISPR-Cas9 genome editing, transforms murine hematopoietic stem cells

Introduction: Pediatric acute megakaryoblastic leukemia is characterized by unique recurrent chromosomal translocations (CTs) with NUP98-KDM 5A being one of the most frequent. However, adequat models to investigate the role of NUP98-KDM 5A in non-DS-AMKL are yet to be established.

Methods: A lentiviral CRISPR-Cas9 vector containing two sgRNAs to generate the Nup98-Kdm5a translocation was generated and introduced into murine lineage^-, Sca1⁺, cKit⁺ (LSK) cells to evaluate the effects of endogenous Nup98-Kdm5a in vitro and in vivo.

Results: Expression of the Nup98-Kdm5a fusion gene was detectable in transduced murine LSK cells resulting in an increased proliferation and self-renewal in vitro. We thereby established a primary cell derived cell line characterized by myeloid morphology and aberrant megakaryocytic cell surface marker (CD41) expression.

Conclusions: Modeling NUP98-KDM 5A via the CRIPSR-Cas9-System allows to study its effects at endogenous expression level. Our model will provide new insights into the role of NUP98-KDMA5A in the transformation towards AMKL. Ongoing in vivo studies will yield novel insights in the role of the microenvironment in AMKL development.

*contributed equally.