Glycan Epitope and Integrin Expression Dynamics Characterize Neural Crest Epithelial-to-Mesenchymal Transition (EMT) in Human Pluripotent Stem Cell Differentiation

Abstract

The neural crest gives rise to progeny as diverse as peripheral neurons, myelinating cells, cranial muscle, bone and cartilage tissues, and melanocytes. Neural crest derivation encompasses complex morphological change, including epithelial-to-mesenchymal transition (EMT) and migration to the eventual target locations throughout the body. Neural crest cultures derived from stem cells provide an attractive source for developmental studies in human model systems, of immediate biomedical relevance for neurocristopathies, neural cancer biology and regenerative medicine, if only appropriate markers for lineage and cell type definition and quality control criteria were available. Implementing a defined, scalable protocol to generate neural crest cells from embryonic stem cells, we identify stage-defining cluster-of-differentiation (CD) surface markers during human neural crest development in vitro. Acquisition of increasingly mesenchymal phenotype was characterized by absence of neuroepithelial stemness markers (CD15, CD133, CD49f) and by decrease of CD57 and CD24. Increased per-cell-expression of CD29, CD44 and CD73 correlated with established EMT markers as determined by immunofluorescence and immunoblot analysis. The further development towards migratory neural crest was associated with decreased CD24, CD49f (ITGA6) and CD57 (HNK1) versus an enhanced CD49d (ITGA4), CD49e (ITGA5) and CD51/CD61 (ITGAV/ITGB3) expression. Notably, a shift from CD57 to CD51/CD61 was identified as a sensitive surrogate surface indicator of EMT in neural crest in vitro development. The reported changes in glycan epitope and integrin surface expression may prove useful for elucidating neural crest stemness, EMT progression and malignancies. Graphical Abstract

Developmentally regulated expression of integrin alpha-6 distinguishes neural crest derivatives in the skin

“…ITGA6 has been identified as a robust marker for stem cells of different sources, including epithelial, neural, hematopoietic, germline, and mesenchymal stem cells, and regulates their biological behaviors through cross-talk with environmental cues ( Krebsbach and Villa-Diaz, 2017 ; Xu et al, 2019 ; Yang et al, 2020 ; Zha et al, 2021 ). The expression of ITGA6 has also been found to negatively affect the migration of neural crest cells and play a role in their subsequent differentiation ( Strachan and Condic, 2003 ; Kim et al, 2014 ; Corallo et al, 2020 ; Thomas et al, 2022 ).…”

TCRαβ+NK1.1-CD4-CD8- double-negative T cells inhibit central and peripheral inflammation and ameliorate ischemic stroke in mice

“…Moreover, the sensorimotor function was also improved after delayed DNT cell administration ( Figure S2 C). CD15 and CD24 hi are considered as the surface markers of neuronal progenitors 21 , 22 . In this study, the proportion of Glast - CD24 hi CD15 + neuronal progenitors in the infarct area was detected by flow cytometry.…”

Lymph Node Metastasis-Related Gene ITGA4 Promotes the Proliferation, Migration, and Invasion of Gastric Cancer Cells by Regulating Tumor Immune Microenvironment

“…In addition, our results show that undifferentiated M0 type TAMs is enriched in tissues with high expression of ITGA4. This means that the high expression of ITGA4 is closely related to macrophage polarization and may induce more TAMs [ 28 ]. In the future, researches should pay more attention to how ITGA4 in the GC microenvironment affects the dynamic polarization of macrophages.…”