Download P004 Controlling mouse embryonic stem cell fate using novel GSK

P004 Controlling mouse embryonic stem cell fate using novel
GSK-3 inhibitors
Yolanda Sanchez-Ripoll, Heather Bone,
Benjamin Kumpfmueller, Teresa Damiano,
Adam Nelson, Melanie J. Welham
Centre for Regenerative Medicine, University of Bath, and
University of Leeds.
Pluripotent embryonic stem cells (ESCs) have great potential for
use in regenerative medicine and drug discovery. However, in
order to harness this potential, we must understand the molecular
mechanisms regulating self-renewal and differentiation. Previous
studies had implicated Glycogen Synthase Kinase-3 (GSK-3) in
both maintenance of pluripotency and neuronal differentiation.
To investigate the role of GSK-3 in control of ESC fate further,
we generated a series of bis-indolylmaleimides that selectively
inhibit GSK-3. We demonstrate that specific bis-indolylmaleimides
enhance self-renewal of murine ESCs in the presence of LIF and
serum but not in the absence of LIF. Furthermore, expression of
known markers of pluripotency such as Nanog, Rex1 and Oct4,
are increased. However, the exact mechanism of action of GSK-3
in enhancing self-renewal is unclear. Early changes in Nanog
protein levels following GSK-3 inhibition, suggest that GSK-3
may control its translation. Reports have also implicated GSK-3
inhibition in the generation of mesendodermal progenitors from
ESCs. Treatment with our GSK-3 inhibitors increased expression
of mesodermal markers such as Brachyury and Nodal, demonstrated using reporter lines and RT-PCR. Our series of selective
GSK-3 inhibitors are proving to be valuable tools for deciphering
the function of GSK-3 in regulating ESC fate.