Nodal/Activin Signaling Predicts Human Pluripotent Stem Cell Lines Prone to Differentiate Toward the Hematopoietic Lineage

Abstract

Lineage-specific differentiation potential varies among different human pluripotent stem cell (hPSC) lines, becoming therefore highly desirable to prospectively know which hPSC lines exhibit the highest differentiation potential for a certain lineage. We have compared the hematopoietic potential of 14 human embryonic stem cell (hESC)/induced pluripotent stem cell (iPSC) lines. The emergence of hemogenic progenitors, primitive and mature blood cells, and colony-forming unit (CFU) potential was analyzed at different time points. Significant differences in the propensity to differentiate toward blood were observed among hPSCs: some hPSCs exhibited good blood differentiation potential, whereas others barely displayed blood-differentiation capacity. Correlation studies revealed that the CFU potential robustly correlates with hemogenic progenitors and primitive but not mature blood cells. Developmental progression of mesoendodermal and hematopoietic transcription factors expression revealed no correlation with either hematopoietic initiation or maturation efficiency. Microarray studies showed distinct gene expression profile between hPSCs with good versus poor hematopoietic potential. Although neuroectoderm-associated genes were downregulated in hPSCs prone to hematopoietic differentiation many members of the Nodal/Activin signaling were upregulated, suggesting that this signaling predicts those hPSC lines with good blood-differentiation potential. The association between Nodal/Activin signaling and the hematopoietic differentiation potential was confirmed using loss- and gain-of-function functional assays. Our data reinforce the value of prospective comparative studies aimed at determining the lineage-specific differentiation potential among different hPSCs and indicate that Nodal/Activin signaling seems to predict those hPSC lines prone to hematopoietic specification.

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

“…Next, we tested whether this prenatal leukemic fusion may impact the hemato-endothelial cell fate of hESCs using in vitro hematopoietic and endothelial differentiation from hESCs as surrogate assays for early developmental events. During human embryoid body (hEB) differentiation, a population of primitive hemogenic precursors arises, which is uniquely responsible for hematopoietic and endothelial development [14,15]. We thus investigated first the effect of MLL-AF4 on the emergence of hemogenic precursors (CD45-CD31 + CD34 + ) throughout hEB development (Figure 2A and 2B).…”

“…H9 and AND-1 hESCs were maintained undifferentiated in a feeder-free culture as previously described [14,39]. Briefly, hESCs were cultured in Matrigel (BD Biosciences, Bedford, MA, USA)-coated T25 flasks in human feeder conditioned medium (CM) supplemented with 8 ng/ml basic fibroblast growth factor (bFGF; Miltenyi, Germany) [40].…”

“…The fact that leukemogenesis manifests as altered cell differentiation suggests that hematopoietic-directed differentiation of hESCs could become a promising humanspecific strategy to study the onset of hematopoiesis, particularly the emergence of the earliest events leading to the specification of both normal and abnormal hematopoietic tissue [12]. During hESC differentiation, a population of primitive hemogenic precursors arises that is uniquely responsible for hematopoietic and endothelial development [13][14][15]. Interestingly, MLL fusions have also been implicated in endothelial cell maturation [16] and endothelial dysfunction has recently been linked to disease outcome in childhood leukemias [17].…”

MSX2 mediates entry of human pluripotent stem cells into mesendoderm by simultaneously suppressing SOX2 and activating NODAL signaling

“…Activation of Wnt signaling results in a loss of pluripotency and drives hPSC differentiation towards endoderm and mesoderm [12][13][14], and activation of BMP signaling leads to mesendoderm or trophoectoderm differentiation, depending on the dose and duration of stimulation [15,16]. Interestingly, Nodal/Activin signaling cooperates with bFGF to maintain pluripotency [17,18], but promotes mesendoderm differentiation of hESCs in the absence of bFGF [19][20][21]. Despite these studies, there are still significant gaps in the knowledge of the signaling mechanisms that regulate cell fate transitions from pluripotency to the embryonic germ layers.…”

Insights into the cellular origin and etiology of the infant pro-B acute lymphoblastic leukemia with MLL-AF4 rearrangement

“…19,27 Therefore, using conditions previously optimized to promote hematopoietic differentiation from hESCs, 16,29,[31][32][33][34][35][36][37][38][39][40][41] Bueno et al assessed to what extent etoposide induces MLL breaks at two different developmental stages during human embryonic hematopoietic development. Early (day þ 15) hESC-derived hematopoietic derivatives seem to be slightly more susceptible to etoposide-induced MLL breaks than late (day þ 22) fully differentiated hESC-derived hematopoietic derivatives (2.9 vs 1.6%, respectively).…”