Brief Report: Ectopic Expression of Nup98-HoxA10 Augments Erythroid Differentiation of Human Embryonic Stem Cells

Abstract

Hox genes encode highly conserved transcription factors that have been implicated in hematopoietic development and self-renewal of hematopoietic stem cells (HSCs) and hematopoietic development. The potency of NUP98-HOXA10hd (NA10) on adult murine bone marrow HSC selfrenewal prompted us to examine its effect on specification and proliferation of hematopoietic cells derived from human embryonic stem cells (hESCs). Here, we demonstrate that expression of NA10 in hESCs influences the hematopoietic differentiation program. The specific effect of NA10 is dependent on the developmental stage of hematopoietic emergence from hESCs. Overexpression of NA10 in either undifferentiated hESCs or early hemogenic precursors augmented the frequency of CD452 GlycophorinA 1 cells and erythroid progenitors (blast-forming unit-erythrocyte). In contrast, targeted NA10 expression in primitive CD341 cells committed to the hematopoietic lineage had no effect on erythropoietic capacity but instead increased hematopoietic progenitor proliferation. Our study reveals a novel neomorphic effect of NA10 in early human erythroid development from pluripotent stem cells.

A human ESC model for MLL-AF4 leukemic fusion gene reveals an impaired early hematopoietic-endothelial specification

“…The fact that leukemogenesis manifests as a blockage or altered cell differentiation suggests that hematopoietic differentiation of hESCs could become a promising human-specific strategy to study the onset of hematopoiesis, particularly the emergence of the earliest events leading to the specification of the hematopoietic tissue [12]. Previous studies made use of the BCR-ABL fusion gene to promote hematopoietic proliferation in mouse ESCs [27,28] and only a very recent study has studied the impact of Nup98-HoxA10 fusion oncogene on hESC-derived hematopoiesis [29]. However, this is the first study exploring the developmental impact of a leukemic fusion known to arise in utero on hESC hemato-endothelial development.…”

“…Long-term, large-scale culture of MLL-AF4 hESC-derived hematopoietic cells or hemogenic precursors provides an unprecedented system for drug screening and toxicity studies. It also offers a unique in vitro system to test the ability of potential cooperating oncogenic events (reciprocal AF4-MLL or FLT3-activating mutations) or causal genotoxic compounds to induce leukemic transformation in vitro, characterized by the outgrowth of malignant Blymphoid/monocyte clones [29,37,38]. Taken …”

Generation and Characterization of Erythroid Cells from Human Embryonic Stem Cells and Induced Pluripotent Stem Cells: An Overview

“…A recently published method of culturing hESCs and hiPSCs in suspension may aid in the scaling up production of hESCs and hiPSCs [40]. In addition, the proposed utilization of synthetic 3D structures mimicking bone marrow structure [41] and the manipulation of transcriptional environment, such as ectopic expression of engineered Nup98-HoxA10 fusion protein [42] and downregulation of microRNAs-126/125* [43], may improve the erythroid production efficiency. …”

Development of Stem Cell Therapy for Medical Uses

“…The choice of tissue depends on ease of access and functional requirements [129][130][131][132][133]. The choice of tissue depends on ease of access and functional requirements [129][130][131][132][133].…”

“…To make transfusion product cost-effective, considerable amount of progress in the expansion, maturation, and terminal differentiation/ enucleation of erythrocytes will be needed [131]. may also help to improve production efficiency [132,133]. Furthermore, utilization of synthetic 3D structures mimicking bone marrow structure and the manipulation of transcriptional environment (e.g., downregulation of microRNAs-126/125*, ectopic expression of engineered Nup98-HoxA10 fusion protein, etc.)…”