Understanding and controlling cell fate decisions

Supervisors: Sally Lowell, Kim Dale

Project description:

The aim of this project is to understand how pluripotent cells of the early embryo reliability select the correct differentiated fate at the appropriate time and place at gastrulation. We will use this knowledge to improve our ability to control differentiation of pluripotent cells (ES cells and iPS cells) in culture.

The global signals that control cell fate at gastrulation are now well understood, yet these signals do not reliably direct differentiation in culture for reasons we do not understand [1].  We will explore the idea that local communication between individual cells also influences differentiation decisions.

The Lowell lab has developed quantitative image analysis tools for following differentiation at single cell resolution in embryos, organoid-type cultures, or conventional monolayer cultures.  These tools will be used to uncover transient local patterning in early cell fate decisions.  These patterns will give clues as to the particular modes of local cell-cell interactions that are operating in these tissues. This will be explored using the expertise in mathematical modelling from the Dale lab.

The project will focus on Notch signalling as a candidate regulator of these local interactions [2,3]. We already know that Notch becomes active in certain regions of the gastrulating embryo and that it is capable of influencing differentiation of pluripotent cells by adjusting responsiveness to instructive signals.  The Lowell and Dale labs have complementary expertise in studying the Notch pathway in a number of different contexts. This project will use a set of tools for monitoring and manipulating Notch that have been developed in both labs in order to uncover the mechanisms by which Notch modulates cell fate decisions.  These findings will help us to understand how local cell-cell interactions confer robustness on developmental patterning in vivo, and provide new approaches for generating useful cell times from ES and iPS cells in culture.

Training outcomes

Wet lab skills: differentiation of pluripotent cells in monolayer and organoid cultures, embryology, genome engineering (routine techniques in Lowell and Dale labs).

Imaging skills: confocal and light-sheet microscopy, use of high-level quantitative image analysis software (already custom written and in use within Lowell lab)

Quantitative and mathematical modelling skills (Dale lab).

References

[1] Malaguti M, Nistor PA, Blin G, Pegg A, Zhou X, Lowell S. Bone morphogenic protein signalling suppresses differentiation of pluripotent cells by maintaining expression of E-Cadherin. Elife. 2013 Dec 17;2:e01197. doi: 10.7554/eLife.01197.

[2] Lowell S, Benchoua A, Heavey B, Smith AG. Notch promotes neural lineage entry  by pluripotent embryonic stem cells. PLoS Biol. 2006 May;4(5)

[3] Carrieri FA, Dale JK. Turn It Down a Notch. Front Cell Dev Biol. 2017 Jan 18;4:151.

 

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