Transcriptional and signaling networks establish complex cross-regulatory interactions that drive cellular differentiation during development. Using microarrays we identified the gene encoding the ligand Wnt9a as a candidate target of Neurogenin3, a basic helix-loop-helix transcription factor that functions as a master regulator of pancreatic endocrine differentiation. Here we show that Wnt9a is expressed in the embryonic pancreas and that its deficiency enhances activation of the endocrine transcriptional program and increases the number of endocrine cells at birth. We identify the gene encoding the endocrine transcription factor Nkx2-2 as one of the most upregulated genes in Wnt9a-ablated pancreases and associate its activation to reduced expression of the Wnt effector Tcf7l2. Accordingly, in vitro studies confirm that Tcf7l2 represses activation of Nkx2-2 by Neurogenin3 and inhibits Nkx2-2 expression in differentiated β-cells. Further, we report that Tcf7l2 protein levels decline upon initiation of endocrine differentiation in vivo, disclosing the downregulation of this factor in the developing endocrine compartment. These findings highlight the notion that modulation of signalling cues by lineage-promoting factors is pivotal for controlling differentiation programs.

This work has been supported by the Spanish Ministerio de Ciencia e Innovación (BFU2008-02299/BMC to RGa), Ministerio de Economía y Competitividad/Instituto de Salud Carlos III (PI13/01500 to RGa) and Generalitat de Catalunya (2014 SGR659 to RGo). Predoctoral fellowships were provided by the Spanish Ministerio de Ciencia e Innovación (BES-2007- 17284, GP) and IDIBAPS (ME). The research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/2009-2013) under the grant agreement n°229673 (SC). CIBERDEM (Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas) is an initiative of the Instituto de Salud Carlos III.