Experimental measurements indicate a second signal mechanism, in addition to canonical WNT signaling, being involved in the regulation of nuclear B-catenin levels during the cell fate commitment phase of neural differentiation.

However, to control hNPC differentiation within the scope of stem cell engineering, a thorough understanding of cell fate determination and its endogenous regulation is required.

Here we investigate the spatio-temporal regulation of WNT/fi-catenin signaling in the process of cell fate commitment in hNPCs, which has been reported to play a crucial role for the differentiation process of hNPCs.

In a combined in-vitro and in-silico approach we find strong evidence, that cell fate commitment in human neural progenitor cells is driven by two distinct fi-catenin signaling mechanisms.

However, controlling NPC differentiation in stem cell engineering demands a thorough understanding of neuronal and glial cell fate determination and its endogenous regulation.

A first characterization of ReNcell VM197 hNPC cell fate commitment uncovered a spa-tio-temporal regulation of WNT/fi-catenin key proteins, like LRP6, DVL, AXIN and fi-catenin throughout the entire phase of early differentiation [8].

However, the exact mechanisms that drive the WNT/fi-catenin signaling and therewith control the cell fate commitment in hNPC remain unclear.

We evaluate the impact of lipid raft disruption on WNT/fi-catenin signaling during differentiation by measuring the temporal progress of WNT signaling in terms of nuclear fi-catenin concentrations in methyl-fi-cyclodextrin-treated and untreated cells in the process of cell fate commitment.

To explore the potential mechanisms that drive the spatio-temporal regulation of fi-catenin signaling during cell fate commitment and to close this important gap in existing models, we built a comprehensive, stochastic WNT/fi-catenin signaling model, that combines both, mem-brane-related and intracellular processes.

This allows us to study WNT signaling in the context of cell fate commitment in a time dependent manner.

Identifying control strategies for biological networks is paramount for practical applications that involve reprogramming a cell’s fate , such as disease therapeutics and stem cell reprogramming.

In biological situations (like in our test cases) one commonly has certain molecular markers of cell fate which specify the attractor to a large degree but not at the level of every node.

The network control framework we propose is applicable to any cell fate reprogramming process for which a logical dynamical model can be constructed.

To demonstrate the potential of our framework, we choose two types of cell fate reprogramming processes: disease therapeutics and cell differentiation.

The framework presented in this work is formulated and applied in the context of logical network modeling of cell fate reprogramming processes but its applicability is not restricted to it.

Practical applications such as stem cell reprogramming [1—3] and the search for new therapeutic targets for diseases [4—6] have also motivated a great interest in the general task of cell fate reprogramming, i.e., controlling the internal state of a cell so that it is driven from an initial state to a final target state (see references [7—13]).

The attractors of intracellular networks have been found to be identifiable with different cell fates , cell behaviors, and stable patterns of cell activity [24—30, 39, 40].

We refer to such a diagram as a stable motif succession diagram, and we note that it is closely analogous to a cell fate decision diagram.

The observation that neuroblastoma cell lines expressed more than half of the RTKs in the human genome (S3 Fig), and responded to signals from growth factors in the embryonic mi-croenvironment to migrate and differentiate into a number of neural crest target sites (S4 Fig), suggests that neuroblastoma, and neural crest from which it is derived, takes full advantage of RTK signaling mechanisms to govern cell fate decisions.

Compartmentalization of signal-initiating receptors and downstream effectors may be employed to distinguish extracellular instructions that determine cell fate .

In fact, there is evidence that endosomal signaling from a number of different receptors affects cell fate decisions during development [62—66].

That neuroblastoma cells express so many RTKs suggests that mechanisms to discern and integrate different receptors’ signals must play a role in cell fate decisions in neural crest and neuroblastoma [106—108].

Neural crest cells appear to restrict their range of cell fate choices in sequential steps [7,8], and the profound heterogeneity in neuroblastoma is caused by a failure to differentiate at different stages.

The immediate-early response mediates cell fate in response to a variety of extracellular stimuli and is dysregulated in many cancers.

The source and duration of the induction signal can determine alternative cell fates , for example, transient signalling may result in cell proliferation, whereas sustained signalling gives rise to cell differentiation [2].

The activation of ErbB receptors by epidermal growth factor (EGF) or heregulin (HRG) in the MCF7 breast cancer cell line exemplifies the impact of such transient or sustained signalling on cell fate [3, 4].

This class of transcripts is currently understudied, but lncRNAs are differentially expressed during differentiation, are preferentially localised in chromatin and have been proposed to ‘f1ne-tune’ cell fate via their roles in transcriptional regulation [14—16].