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Human pluripotent stem cell modeling, from normal development to neuroblastoma

(2024)
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Abstract
Understanding the nuances of human NCC development and their perturbations is crucial for comprehending neural crest disorders, particularly NB, the most prevalent extra-cranial solid tumor affecting children. Faithfully recapitulating in vivo developmental processes using in vitro differentiation of ESC has long been challenge. However, using single-cell transcriptome analysis we could confirm that our model was able to generate hSAP developmental subpopulations that transcriptionally resembled their in vivo counterparts. We further confirmed that our developing cells largely mirrored the nuanced intricacies witnessed during in vivo human development, offering a unique window into the complexities of NCC development and providing a solid foundation for future investigations into both normal and aberrant human neural crest development. As a proof of concept we used our model to investigate the impact of the ALKR1275Q mutation, commonly observed familial NB, on the normal development. This analysis revealed a distinct subpopulation, characterized by a marked increase in SOX2 expression, a factor intricately associated with preserving an undifferentiated cellular state. Therefore, we hypothesize that constitutive ALK signaling may cause aberrant expression of SOX2 and its target genes in a subset of NCC, thereby leading to a differentiation blockage and rendering these cells more vulnerable to subsequent mutations. Additionally, we explored the effect of MYCN overexpression in the hSAP developmental window. We could observe that overexpression of MYCN during the hSAP developmental phase, results in a substantial wave of cell death, particularly affecting bridging cells and CPCs and ECCs. The surviving cells exhibit a distinctive transcriptional response characteristic of cancer hallmarks, including evasion of apoptosis and a remarkable surge in cellular proliferation, all the while preserving their developmental identities. Further experiments with Xenografts were able to show that that the surviving cells, subjected to MYCN overexpression, possess the capacity to form tumors, however, display transcriptional variability contingent on the timing of induction of MYCN overexpression. These insights enhance our comprehension of the intricate role of MYCN in NB pathogenesis and tumorigenesis, opening new avenues for therapeutic interventions and treatments. Our work emphasizes the potential of our in vitro developmental model to delve into the complexities of SAP development and its perturbations. Understanding the impact of genetic aberrations, such as ALK and MYCN, provides insights into NB pathogenesis and potential therapeutic interventions, setting the stage for further investigations into the development and treatment of NB.
Keywords
In vitro modeling, neuroblastoma, ALK, MYCN, sympatho-adrenergic development

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@phdthesis{01HT5903H3WJCNTWCVPNE4KY2J,
  abstract     = {{Understanding the nuances of human NCC development and their perturbations is crucial for comprehending neural crest disorders, particularly NB, the most prevalent extra-cranial solid tumor affecting children. Faithfully recapitulating in vivo developmental processes using in vitro differentiation of ESC has long been challenge. However, using single-cell transcriptome analysis we could confirm that our model was able to generate hSAP developmental subpopulations that transcriptionally resembled their in vivo counterparts. We further confirmed that our developing cells largely mirrored the nuanced intricacies witnessed during in vivo human development, offering a unique window into the complexities of NCC development and providing a solid foundation for future investigations into both normal and aberrant human neural crest development. 

As a proof of concept we used our model to investigate the impact of the ALKR1275Q mutation, commonly observed familial NB, on the normal development.  This analysis revealed a distinct subpopulation, characterized by a marked increase in SOX2 expression, a factor intricately associated with preserving an undifferentiated cellular state. Therefore, we hypothesize that constitutive ALK signaling may cause aberrant expression of SOX2 and its target genes in a subset of NCC, thereby leading to a differentiation blockage and rendering these cells more vulnerable to subsequent mutations. Additionally, we explored the effect of MYCN overexpression in the hSAP developmental window. We could observe that overexpression of MYCN during the hSAP developmental phase, results in a substantial wave of cell death, particularly affecting bridging cells and CPCs and ECCs. The surviving cells exhibit a distinctive transcriptional response characteristic of cancer hallmarks, including evasion of apoptosis and a remarkable surge in cellular proliferation, all the while preserving their developmental identities. Further experiments with Xenografts were able to show that that the surviving cells, subjected to MYCN overexpression, possess the capacity to form tumors, however, display transcriptional variability contingent on the timing of induction of MYCN overexpression. These insights enhance our comprehension of the intricate role of MYCN in NB pathogenesis and tumorigenesis, opening new avenues for therapeutic interventions and treatments.
Our work emphasizes the potential of our in vitro developmental model to delve into the complexities of SAP development and its perturbations. Understanding the impact of genetic aberrations, such as ALK and MYCN, provides insights into NB pathogenesis and potential therapeutic interventions, setting the stage for further investigations into the development and treatment of NB.}},
  author       = {{Van Haver, Stéphane}},
  keywords     = {{In vitro modeling,neuroblastoma,ALK,MYCN,sympatho-adrenergic development}},
  language     = {{eng}},
  pages        = {{273}},
  publisher    = {{Ghent University. Faculty of Medicine and Health Sciences}},
  school       = {{Ghent University}},
  title        = {{Human pluripotent stem cell modeling, from normal development to neuroblastoma}},
  year         = {{2024}},
}