Establishment and controlled differentiation of neural crest stem cell lines using conditional transgenesis

Maurer, Jochen; Fuchs, Sebastian; Jäger, Richard; Kurz, Bodo; Sommer, Lukas; Schorle, Hubert

doi:10.1111/j.1432-0436.2007.00164.x

Cited by 54 publications

(65 citation statements)

References 57 publications

(79 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To further test the cell-autonomous role of Brg1 and develop cellular assays to study Brg1 biology in NCCs, we knocked down Brg1 in Joma1.3 cells, a transformed murine NCC line that expresses early NCC markers and can be induced to differentiate into SMCs (20). We generated stable Joma1.3 NCCs by infecting these cells with lentivirus that allowed doxycycline (Dox)-inducible expression of shRNA against Brg1 (designated siBrg1 NCCs), as well as a scrambled control shRNA (designated siCtrl NCCs) ( Fig.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Brg1 governs distinct pathways to direct multiple aspects of mammalian neural crest cell development

Xiong²,

Shang³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Development of the cerebral vessels, pharyngeal arch arteries (PAAs). and cardiac outflow tract (OFT) requires multipotent neural crest cells (NCCs) that migrate from the neural tube to target tissue destinations. Little is known about how mammalian NCC development is orchestrated by gene programming at the chromatin level, however. Here we show that Brahma-related gene 1 (Brg1), an ATPase subunit of the Brg1/Brahma-associated factor (BAF) chromatin-remodeling complex, is required in NCCs to direct cardiovascular development. Mouse embryos lacking Brg1 in NCCs display immature cerebral vessels, aberrant PAA patterning, and shortened OFT. Brg1 suppresses an apoptosis factor, Apoptosis signal-regulating kinase 1 (Ask1), and a cell cycle inhibitor, p21 cip1 , to inhibit apoptosis and promote proliferation of NCCs, thereby maintaining a multipotent cell reservoir at the neural crest. Brg1 also supports Myosin heavy chain 11 (Myh11) expression to allow NCCs to develop into mature vascular smooth muscle cells of cerebral vessels. Within NCCs, Brg1 partners with chromatin remodeler Chromodomain-helicase-DNAbinding protein 7 (Chd7) on the PlexinA2 promoter to activate PlexinA2, which encodes a receptor for semaphorin to guide NCCs into the OFT. Our findings reveal an important role for Brg1 and its downstream pathways in the survival, differentiation, and migration of the multipotent NCCs critical for mammalian cardiovascular development.N eural crest cells (NCCs) originate from the neural crest of the dorsal neural tube and migrate to many regions of the embryo, where they differentiate into a variety of local cells, including cardiovascular tissues (1). NCCs that emigrate from the neural crest of rhombomere 6-8 to pharyngeal arches and the heart are essential for the patterning of pharyngeal arch arteries (PAAs) and the cardiac outflow tract (OFT) (2, 3). These NCCs also differentiate into vascular smooth muscle cells (SMCs) of PAAs and the muscular septum of the aorta and pulmonary trunk (4, 5). In contrast, NCCs from the cephalic neural tube migrate to the face and forebrain to form craniofacial bones, as well as SMCs of facial and forebrain vessels (6). Thus, NCCs are critical for the formation of cardiac OFT and vascular supplies of large areas of the body.Disruption of NCC development, either directly or indirectly, results in many forms of human birth defects with cardiovascular malformations, including Alagille, Carpenter, Ivemark, Leopard, Williams, DiGeorge, and CHARGE syndromes (7). These syndromes involve defects in PAAs or cardiac OFT, such as coarctation of the aorta, interrupted aortic arch, pulmonary artery stenosis, double-outlet right ventricle, tetralogy of Fallot, or persistent truncus arteriosus. During PAA and OFT development, NCCs are regulated by numerous transcription factors, including Pax3, Pbx1/2/3, Tbx1/2/3/20, Msx1/2, Hand2, AP2a, Cited2, Pitx2, Sox4, Foxc1/c2/d3/h1, Fog2, Gata3/4/6, and Notch/NICD (8). Such extensive involvement of transcription factors indicates the importance of gene pr...

show abstract

Section: Resultsmentioning

confidence: 99%

“…To test the effect of Brg1 on the differentiation of NCCs into SMCs, we treated siBrg1 NCCs with Tgfβ to induce their differentiation into mature SMCs expressing Myh11 (20). qRT-PCR showed that Tgfβ induced Myh11 expression in siCtrl NCs, but not in siBrg1 NCCs (Fig.…”

Section: Resultsmentioning

confidence: 99%

Brg1 governs distinct pathways to direct multiple aspects of mammalian neural crest cell development

Xiong²,

Shang³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…JoMa1 cells were cultured in a modified neural crest culture medium (NCC medium; Maurer et al, 2007). Dulbecco's modified Eagle's medium (4,5 mg/ml glucose, L-glutamine, Pyruvate) and Ham's F12 medium were mixed 1:1 and supplemented with 1% N2-Supplement (Invitrogen), 2% B27-Supplement (Invitrogen), 10 ng/ml epidermal growth factor (PromoKine, Heidelberg, Germany), 1 ng/ml FGF (Millipore, Billerica, MA, USA), 100 U/ml Penicillin-Streptomycin (Invitrogen) and 10% chick embryo extract (Pajtler et al, 2010).…”

Section: Joma1 Cell Experimentsmentioning

confidence: 99%

Oncogenic activation of FOXR1 by 11q23 intrachromosomal deletion-fusions in neuroblastoma

et al. 2011

View full text Add to dashboard Cite

Neuroblastoma tumors frequently show loss of heterozygosity of chromosome 11q with a shortest region of overlap in the 11q23 region. These deletions are thought to cause inactivation of tumor suppressor genes leading to haploinsufficiency. Alternatively, micro-deletions could lead to gene fusion products that are tumor driving. To identify such events we analyzed a series of neuroblastomas by comparative genomic hybridization and single-nucleotide polymorphism arrays and integrated these data with Affymetrix mRNA profiling data with the bioinformatic tool R2 (http://r2.amc.nl). We identified three neuroblastoma samples with small interstitial deletions at 11q23, upstream of the forkhead-box R1 transcription factor (FOXR1). Genes at the proximal side of the deletion were fused to FOXR1, resulting in fusion transcripts of MLL-FOXR1 and PAFAH1B2-FOXR1. FOXR1 expression has only been detected in early embryogenesis. Affymetrix microarray analysis showed high FOXR1 mRNA expression exclusively in the neuroblastomas with micro-deletions and rare cases of other tumor types, including osteosarcoma cell line HOS. RNAi silencing of FOXR1 strongly inhibited proliferation of HOS cells and triggered apoptosis. Expression profiling of these cells and reporter assays suggested that FOXR1 is a negative regulator of fork-head box factormediated transcription. The neural crest stem cell line JoMa1 proliferates in culture conditional to activity of a MYC-ER transgene. Over-expression of the wild-type FOXR1 could functionally replace MYC and drive proliferation of JoMa1. We conclude that FOXR1 is recurrently activated in neuroblastoma by intrachromosomal deletion/fusion events, resulting in overexpression of fusion transcripts. Forkhead-box transcription factors have not been previously implicated in neuroblastoma pathogenesis. Furthermore, this is the first identification of intrachromosomal fusion genes in neuroblastoma.

show abstract

“…Joma1.3 cells were cultured as described (Maurer et al, 2007). Briefly, cells were plated on cell culture dishes coated with 1 mg/ml fibronectin (Roche) in a 1:1 mixture of Dulbecco's modified Eagle's medium and F12 medium (Gibco) containing 1% N2 supplement (Invitrogen), 2% B27 supplement (Invitrogen), 10 ng/ml epidermal growth factor (Invitrogen), 1 ng/ml fibroblast growth factor (Invitrogen), 100 U/ml penicillin/ streptomycin (Invitrogen) and 10% chick embryo extract (Stemple and Anderson, 1992).…”

Section: Joma13 Cell Culturementioning

confidence: 99%

The neural crest-enriched microRNA miR-452 regulates epithelial-mesenchymal signaling in the first pharyngeal arch

et al. 2010

View full text Add to dashboard Cite

SUMMARYNeural crest cells (NCCs) are a subset of multipotent, migratory stem cells that populate a large number of tissues during development and are important for craniofacial and cardiac morphogenesis. Although microRNAs (miRNAs) have emerged as important regulators of development and disease, little is known about their role in NCC development. Here, we show that loss of miRNA biogenesis by NCC-specific disruption of murine Dicer results in embryos lacking craniofacial cartilaginous structures, cardiac outflow tract septation and thymic and dorsal root ganglia development. Dicer mutant embryos had reduced expression of Dlx2, a transcriptional regulator of pharyngeal arch development, in the first pharyngeal arch (PA1). miR-452 was enriched in NCCs, was sufficient to rescue Dlx2 expression in Dicer mutant pharyngeal arches, and regulated non-cell-autonomous signaling involving Wnt5a, Shh and Fgf8 that converged on Dlx2 regulation in PA1. Correspondingly, knockdown of miR-452 in vivo decreased Dlx2 expression in the mandibular component of PA1, leading to craniofacial defects. These results suggest that posttranscriptional regulation by miRNAs is required for differentiation of NCC-derived tissues and that miR-452 is involved in epithelial-mesenchymal signaling in the pharyngeal arch.

show abstract

Establishment and controlled differentiation of neural crest stem cell lines using conditional transgenesis

Cited by 54 publications

References 57 publications

Brg1 governs distinct pathways to direct multiple aspects of mammalian neural crest cell development

Brg1 governs distinct pathways to direct multiple aspects of mammalian neural crest cell development

Oncogenic activation of FOXR1 by 11q23 intrachromosomal deletion-fusions in neuroblastoma

The neural crest-enriched microRNA miR-452 regulates epithelial-mesenchymal signaling in the first pharyngeal arch

Contact Info

Product

Resources

About