Transcriptome profiling of FGL2-stimulated enteric neural crest cells to investigate metabolic dysregulation in Hirschsprung disease pathogenesis
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ABSTRACT:
PROVIDER: PRJNA1304459 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:This dataset comprises bulk RNA sequencing data from primary mouse enteric neural crest cells (ENCCs) stimulated with fibrinogen-like protein 2 (FGL2) or left untreated as controls. ENCCs were isolated from embryonic day 14–15 C57BL/6J mice and cultured under defined conditions to ensure >95% expression of P75 and nestin markers. Following FGL2 stimulation, total RNA was extracted, and sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina. Paired-end sequencing (2 × 150 bp) was performed on the Illumina NovaSeq 6000 platform. The study aims to identify transcriptional changes and signaling pathway alterations induced by FGL2 in ENCCs, providing insights into the role of FGL2-mediated metabolic dysregulation in the pathogenesis of Hirschsprung disease.
2025-08-14 | GSE305114 | GEO
Project description:Background & Aims Enteric glial cells (EGCs) play vital roles in gut homeostasis and neurogenesis. Hirschsprung disease (HSCR) is a neurocristopathy, yet paradoxically, neural crest-derived enteric glial cells (EGCs) are present within the muscle of the affected region. This study investigates the molecular identity, origins, and neurogenic potential of EGCs in the aganglionic mouse and human colon. Methods We utilized single-cell RNA sequencing (scRNA-seq) from aganglionic and ganglionated segments of 10-14 day old Ednrb-null mice (Plp1-GFP;Ednrb-/-) and wild-type controls. Results scRNA-seq revealed the absence of GFAP+ intraganglionic (IG) glia in aganglionic colon, while CAMK2b+ extraganglionic (EG) glia and Schwann-like cells (SLCs) were present. EG glia exhibited a transcriptional profile similar to SLCs, suggesting a possible shared embryonic origin. Conclusions A subpopulation of extraganglionic EGCs, transcriptionally similar to SLCs, persists in the aganglionic segment of HSCR.
2025-10-15 | GSE293138 | GEO
Project description:This study was undertaken to identify gene expression changes in enteric neurons of Hirschsprung disease mice. Methods: We performed single-cell RNA sequencing of enteric nervous system cells isolated from the small intestine of Ednrb-knockout mice at age P14. Results: Through comparisons with published datasets of WT small intestine enteric neurons, we identify a missing neuronal population in the ganglionated small intestine of Hirschsprung disease mice. Conclusion: Single-cell RNA sequencing permits identification of cellular perturbations in Hirschsprung disease models.
2022-08-24 | GSE199683 | GEO
Project description:Enteric nervous system (ENS) development requires migration, proliferation, and neuronal diversification of progenitors to achieve normal gastrointestinal motility. Sox10 deficits disrupt the ratios of mature enteric neuron subtypes. How Sox10 deficiency alters the proportions of neuronal subtypes is still being determined. Sox10's prominent expression in neural crest-derived enteric progenitors and lack of this gene in enteric neurons led us to investigate how Sox10 effects in early enteric progenitors could alter neuronal allocation. To explore this further, enteric progenitors, developing neurons, and enteric glia were isolated from Sox10+/+ and Sox10Dom/+ littermates at 15.5 days post-coitus based on dual Sox10-H2BVenus and Phox2b-H2BCerulean transgene expression for single-cell RNA sequencing (scRNA-seq). The data was processed in R to identify cell-specific markers, differentially expressed genes, biological processes associated with gene ontology, cell fate trajectories, and gene regulatory network activity between genotypes. Hybridization chain reaction (HCR) validated expression changes detected in scRNA-seq.
2025-08-25 | GSE262898 | GEO
Project description:Despite extensive genetic heterogeneity, 72% of pathogenic alleles for Hirschsprung disease (HSCR) arise from coding and regulatory variants in genes of the RET and EDNRB gene regulatory network (GRN) in the enteric nervous system (ENS). Reduced signaling of these two receptors below a threshold in enteric neural crest-derived cells (ENCDCs) leads to a molecular tipping point at which otherwise lesser cellular defects result in aganglionosis. To elucidate the mechanisms leading to enteric neuronal loss from these genetic defects, we generated four strains of mice carrying reduced function alleles at Ret or Ednrb or both, along with their wildtype alleles. ENS tissue- and single-cell gene expression profiling of the developing and postnatal gastrointestinal tract in five mouse models, with various combinations of mutant alleles, revealed 3 major insights: (i) Ret and Ednrb deficiency, rather than complete loss, is sufficient to induce HSCR, (ii) Ret and Ednrb demonstrate strong trans interactions, and (iii) disruption of this interaction leads to cellular fate changes to compensate for neuronal loss. This study of targeted mouse models of a multifactorial disorder reveals how increasing dosage of genetic defects within a GRN leads to quantifiably increasing dysregulation from genotype to gene expression to cellular identity to function. Importantly, our studies establish that aganglionosis results only with severely reduced gene expression at both receptor genes and their consequent disruption of normal and compensatory cell fate trajectories.
2025-04-01 | GSE255963 | GEO
Project description:Despite extensive genetic heterogeneity, 72% of pathogenic alleles for Hirschsprung disease (HSCR) arise from coding and regulatory variants in genes of the RET and EDNRB gene regulatory network (GRN) in the enteric nervous system (ENS). Reduced signaling of these two receptors below a threshold in enteric neural crest-derived cells (ENCDCs) leads to a molecular tipping point at which otherwise lesser cellular defects result in aganglionosis. To elucidate the mechanisms leading to enteric neuronal loss from these genetic defects, we generated four strains of mice carrying reduced function alleles at Ret or Ednrb or both, along with their wildtype alleles. ENS tissue- and single-cell gene expression profiling of the developing and postnatal gastrointestinal tract in five mouse models, with various combinations of mutant alleles, revealed 3 major insights: (i) Ret and Ednrb deficiency, rather than complete loss, is sufficient to induce HSCR, (ii) Ret and Ednrb demonstrate strong trans interactions, and (iii) disruption of this interaction leads to cellular fate changes to compensate for neuronal loss. This study of targeted mouse models of a multifactorial disorder reveals how increasing dosage of genetic defects within a GRN leads to quantifiably increasing dysregulation from genotype to gene expression to cellular identity to function. Importantly, our studies establish that aganglionosis results only with severely reduced gene expression at both receptor genes and their consequent disruption of normal and compensatory cell fate trajectories.
2025-04-01 | GSE227574 | GEO
Project description:Here, human pluripotent stem cells (hiPSCs and hESCs) were induced to differentiate to enteric neurons through neural crest specification. We sequenced mRNA samples from enteric neuronal differentiation of human pluripotent stem cells at 4 different stage to generate the gene expression profiles of these cells.
2017-06-14 | GSE79928 | GEO
Project description:The enteric nervous system (ENS), which is derived from enteric neural crest cells (ENCCs) during gut development, represents the neuronal innervation of the gastrointestinal tract and is critical for regulating normal intestinal function. Compromised ENCC migration can lead to Hirschsprung Disease, which is characterized by an aganglionic distal bowel. We find that removal of the ceca, a paired structure present at the midgut-hindgut junction in avian intestine, leads to severe hindgut aganglionosis, suggesting that the ceca are required for ENS development. To test this, we replaced the ceca of embryonic day 6 (E6) wild-type chicks with ceca from transgenic GFP chicks. Interestingly, the entire hindgut ENS arises from the GFP+ ceca-derived ENCC population. Comparative transcriptome profiling of the cecal buds compared to the interceca region shows that the non-canonical Wnt signaling pathway is preferentially expressed within the ceca. Specifically, Wnt11 is highly expressed in the ceca, as confirmed by RNA in situ hybridization, leading us to hypothesize that cecal expression of Wnt11 is important for ENCC colonization of the hindgut. Organ cultures were prepared using E6 avian intestine, when ENCCs are migrating through the ceca, and showed that Wnt11 inhibits enteric neuronal differentiation. These results reveal an essential role for the ceca during hindgut ENS formation and highlight an important function for non-canonical Wnt signaling in regulating ENCC differentiation and thereby promoting their migration into the colon.
2021-11-01 | GSE182783 | GEO
Project description:Enteric nervous system (ENS) development relies on intestinal colonization by enteric neural crest-derived cells (ENCDCs). This is driven by a population of highly migratory and proliferative cells at the wavefront, but the unique molecular characteristics of the wavefront are unknown. ENCDCs from the wavefront and the trailing region proximal were isolated from 11.5 days post coitum (dpc) Wnt1-tdT mouse intestine and subjected to RNA-seq. Wavefront ENCDCs were transcriptionally distinct from trailing ENCDCs, and temporal modelling confirmed their relative immaturity. Wavefront ENCDCs exhibited altered expression of ECM and cytoskeletal genes, consistent with a migratory phenotype. Unlike postnatal enteric neural stem cells, wavefront ENCDCs possessed characteristics of cells undergoing epithelial-mesenchymal transition and lacked neuronal or glial commitment.
2023-02-15 | GSE217757 | GEO
Project description:During the later stages of enteric nervous system (ENS) development, enteric neural crest derived cells (ENCDC) that have colonized the bowel must complete differentiating and mature into neurons and glia. This process is controlled by a complex array of intrinsic and extrinsic factors. We used microarrays to dermine which genes were differntially expressed in ENCDC versus other cells in the developing bowel. We identified many geness enriched in ENCDC with potential roles in the later stages of ENS development
2018-10-09 | GSE120936 | GEO