FoxP1 promotes midbrain identity in embryonic stem cell‐derived dopamine neurons by regulating Pitx3

Abstract

J. Neurochem. (2010) 113, 836–847. Abstract The robust generation of midbrain dopamine neurons from embryonic stem cells and patient‐specific induced pluripotent stem cells is a prospective tool for the development of new drugs and cell based therapies, and investigations into the aetiology of Parkinson’s disease. To achieve this, it is crucial to identify the fate‐determining regulatory factors that influence dopamine cell fate decision and the underlying molecular machinery. We identified FoxP1 as a novel marker for midbrain dopamine neurons. Enforced expression of FoxP1 in embryonic stem cells actuates the expression of Pitx3, a homeobox protein that is exclusively expressed in midbrain dopaminergic neurons and is required for their differentiation and survival during development and from embryonic stem cells in vitro. We show that FoxP1 can be recruited to the Pitx3 locus in embryonic stem cells and regulate Pitx3 promoter activity in a dual‐luciferase assay. This transcriptional regulation of Pitx3 by FoxP1 depends on the presence of two high affinity binding sites in the distal Pitx3 promoter, through which FoxP1 directly binds as demonstrated by chromatin immunoprecipitation and electrophoretic mobility shift assay. Thus, this study demonstrates for the first time a transcription regulatory role for FoxP1 on the Pitx3 gene in mammalian stem cells.

The distinct and overlapping phenotypic spectra of FOXP1 and FOXP2 in cognitive disorders

“…On the other hand, the fact that CNTNAP2 is implicated in both ASD (Alarcon et al 2008) and in language impairment without ASD (Vernes et al 2008) suggests distinct CNTNAP2 pathways exist. Elsewhere, Foxp1 has also been shown to regulate Pitx3 transcription during the differentiation of ES cells into midbrain dopaminergic neurons (Konstantoulas et al 2010). …”

Temporally controlled modulation of FGF/ERK signaling directs midbrain dopaminergic neural progenitor fate in mouse and human pluripotent stem cells

“…Stimulation of ESC-derived neural progenitors with Shh and FGF8 is used by almost all dopamine differentiation protocols (Barberi et al, 2003;Kim et al, 2002;Lee et al, 2000;Perrier et al, 2004;Yan et al, 2005). However, unless combined with genetic manipulation of mDA transcription factors, such as Pitx3 or Lmx1a (Andersson et al, 2006;Chung et al, 2002;Konstantoulas et al, 2010;Maxwell et al, 2005), the midbrain regional identity of the dopamine neurons generated has remained uncertain. Furthermore, the yield of Th + neurons has often proved unreliable between experiments and even highly variable between different microscopic fields within a single culture.…”

“…Recent years have witnessed a rapid advancement in understanding the regulatory cascade that governs ventral midbrain neuroepithelial cell fate specification and the differentiation and maintenance of mDA neurons (Andersson et al, 2006;Ferri et al, 2007;Nakatani et al, 2009). Overexpression of mDA transcription factors offers a valid strategy for generating DA neurons from embryonic stem cells (ESCs) (Andersson et al, 2006;Chung et al, 2005;Kittappa et al, 2007;Konstantoulas et al, 2010;Maxwell et al, 2005). However, mDA specification via genetic manipulation is limited and context dependent (Andersson et al, 2006;Friling et al, 2009;Parmar and Li, 2007;Roybon et al, 2008).…”

Doublesex and mab-3–related transcription factor 5 promotes midbrain dopaminergic identity in pluripotent stem cells by enforcing a ventral-medial progenitor fate

“…Examination of intrinsic transcriptional regulators and extrinsic niche factors critical for mDA development in ESC differentiation has proved to be a powerful means for illuminating or validating their functions in dopaminergic neuron fate specification (6,(16)(17)(18)(19)(20). Notably, engineered expression of mDA transcription factors such as Lmx1a and Pitx3 induced midbrain regional character in derived dopamine neurons (6,19).…”