Ets Factors Regulate Neural Stem Cell Depletion and Gliogenesis in Ras Pathway Glioma

Breunig, Joshua J.; Levy, Rachelle; Antonuk, C. Danielle; Molina, Jéssica; Dutra‐Clarke, Marina; Park, Hannah; Akhtar, Aslam Abbasi; Kim, Gi Bum; Hu, Xin; Bannykh, Serguei; Verhaak, Roel G.W.; Danielpour, Moise

doi:10.1016/j.celrep.2015.06.012

Cited by 55 publications

(40 citation statements)

References 55 publications

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“…Etv4 and Etv5 are well-known effectors of the FGF-Ras-ERK signaling cascade (Mao et al, 2009; Zhang et al, 2009; Hollenhorst et al, 2011; Li et al, 2012; Breunig et al, 2015). Moreover, ETV5 is specifically expressed in bRG among human cortical progenitor cells (Pollen et al, 2015; Thomsen et al, 2016).…”

Section: Resultsmentioning

confidence: 99%

Analogous mechanism regulating formation of neocortical basal radial glia and cerebellar Bergmann glia

Heng

Guo

Leung

et al. 2017

eLife

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Neocortical basal radial glia (bRG) and cerebellar Bergmann glia (BG) are basal progenitors derived from ventricular apical radial glia (aRG) that selectively lose their apical processes. bRG and BG have been implicated in the expansion and folding of the cerebrum and cerebellum, respectively. Here, we analyzed the molecular characteristics and development of bRG and BG. Transcriptomic comparison revealed striking similarity of the molecular features of bRG and BG. We found that heightened ERK signaling activity in aRG is tightly linked to the temporal formation and the relative abundance of bRG in human and mouse cortices. Forced activation of an FGF-ERK-ETV axis that is crucial to BG induction specifically induced bRG with canonical human bRG features in mice. Therefore, our data point to a common mechanism of bRG and BG generation, bearing implications to the role for these basal progenitors in the evolution of cortical folding of the cerebrum and cerebellum.DOI: http://dx.doi.org/10.7554/eLife.23253.001

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Section: Resultsmentioning

confidence: 99%

Analogous mechanism regulating formation of neocortical basal radial glia and cerebellar Bergmann glia

Heng

Guo

Leung

et al. 2017

eLife

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“…The first plasmid constitutively expresses the pBase integrase enzyme (Figure 1A1. pBase catalyzes the random stable integration of plasmids that are flanked with piggyBac terminal repeats (PB TR) (Breunig et al, 2015; Chen and LoTurco, 2012). The second plasmid constitutively expresses a transactivator protein; the rtTA-V10 (tetON) transactivator protein is shown here (Figure 1A2).…”

Section: Basic Protocol 1: Cloning Of Respective Goi (Genetic Elementmentioning

confidence: 99%

“…For these reasons, investigation of gene function in these diverse contexts requires increasingly precise control over transgene expression. Further, pB-tet-GOI uses piggyBac transposition as a means of genomic integration of transgenic elements, allowing for the stable transduction of cell lineages through non-viral means (Breunig et al, 2015; Chen and LoTurco, 2012). Herein, we provide a detailed protocol for the use of the system in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

Tetracycline‐Inducible and Reversible Stable Gene Expression in Human iPSC‐Derived Neural Progenitors and in the Postnatal Mouse Brain

Akhtar

Breunig

2017

CP Stem Cell Biology

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The pB-tet-GOI plasmid system allows for stable piggyBac transposition-mediated integration into cells, a fluorescent nuclear reporter to identify cells that have been transfected, and robust transgene activation or suppression upon the addition of dox to the cell culture or diet of the animal. Furthermore, the addition of luciferase downstream of the target gene allows for quantitative assessment of gene activity in a non-invasive manner. The protocols herein provide instructions for the use of this system in cell lines as well as in the neonatal mouse brain. Specifically, a detailed protocol is provided to illustrate the: (1) cloning of respective GOI (genetic element of interest) [Basic Protocol 1], (2) nucleofection of the plasmid system into human induced pluripotent stem cell (iPSC)-derived neural progenitors [Basic Protocol 2]; (3) dox-induced activation in vitro or in vivo [Basic Protocol 3]; and (4) the non-invasive assessment of gene activity in vivo by bioluminescence imaging [Basic Protocol 4].

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“…Wiesner et al showed that the triple combination of NRAS‐G12V, EGFRvIII, and shp53 induced gliomas with 100% efficiency and median survival was 83 days. The piggyBac transposon system has been used to induce glioma with HRAS and AKT mutations . One limitation of the existing transposon‐based glioma models is that RAS mutations are rare in human gliomas; thus, there is a need for a highly penetrant glioma mouse model that genetically mimics human gliomas.…”

Section: Introductionmentioning

confidence: 99%

Spontaneous development of intratumoral heterogeneity in a transposon‐induced mouse model of glioma

2018

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Glioma is the most common form of malignant brain cancer in adults. The Sleeping Beauty (SB) transposon‐based glioma mouse model allows for effective in vivo analysis of candidate genes. In the present study, we developed a transposon vector that encodes the triple combination of platelet‐derived growth factor subunit A (PDGFA), and shRNAs against Nf1 and Trp53 (shNf1/shp53). Initiation and progression of glioma in the brain were monitored by expression of a fluorescent protein. Transduction of the vector into neural progenitor and stem cells (NPC) in the subventricular zone (SVZ) of the neonatal brain induced proliferation of oligodendrocyte precursor cells, and promoted formation of highly penetrant malignant gliomas within 2‐4 months. Cells isolated from the tumors were capable of forming secondary tumors. Two transposon vectors, encoding either PDGFA or shNf1/shp53 were co‐electroporated into NPC. Cells expressing PDGFA or shNf1/shp53 were labeled with unique fluorescent proteins allowing visualization of the spatial distribution of cells with different genetic alterations within the same tumor. Tumor cells located at the center of tumors expressed PDGFA at higher levels than those located at the periphery, indicating that intratumoral heterogeneity in PDGFA expression levels spontaneously developed within the same tumor. Tumor cells comprising the palisading necrosis strongly expressed PDGFA, suggesting that PDGFA signaling is involved in hypoxic responses in glioma. The transposon vectors developed are compatible with any genetically engineered mouse model, providing a useful tool for the functional analysis of candidate genes in glioma.

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Ets Factors Regulate Neural Stem Cell Depletion and Gliogenesis in Ras Pathway Glioma

Cited by 55 publications

References 55 publications

Analogous mechanism regulating formation of neocortical basal radial glia and cerebellar Bergmann glia

Analogous mechanism regulating formation of neocortical basal radial glia and cerebellar Bergmann glia

Tetracycline‐Inducible and Reversible Stable Gene Expression in Human iPSC‐Derived Neural Progenitors and in the Postnatal Mouse Brain

Spontaneous development of intratumoral heterogeneity in a transposon‐induced mouse model of glioma

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