Stable expression of neurogenin 1 induces LGR5, a novel stem cell marker, in an immortalized human neural stem cell line HB1.F3.

Abstract

Neural stem cells (NSC) with self-renewal and multipotent properties serve as an ideal cell source for transplantation to treat spinal cord injury, stroke, and neurodegenerative diseases. To efficiently induce neuronal lineage cells from NSC for neuron replacement therapy, we should clarify the intrinsic genetic programs involved in a time- and place-specific regulation of human NSC differentiation. Recently, we established an immortalized human NSC clone HB1.F3 to provide an unlimited NSC source applicable to genetic manipulation for cell-based therapy. To investigate a role of neurogenin 1 (Ngn1), a proneural basic helix-loop-helix (bHLH) transcription factor, in human NSC differentiation, we established a clone derived from F3 stably overexpressing Ngn1. Genome-wide gene expression profiling identified 250 upregulated genes and 338 downregulated genes in Ngn1-overexpressing F3 cells (F3-Ngn1) versus wild-type F3 cells (F3-WT). Notably, leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5), a novel stem cell marker, showed an 167-fold increase in F3-Ngn1, although transient overexpression of Ngn1 did not induce upregulation of LGR5, suggesting that LGR5 is not a direct transcriptional target of Ngn1. KeyMolnet, a bioinformatics tool for analyzing molecular relations on a comprehensive knowledgebase, suggests that the molecular network of differentially expressed genes involves the complex interaction of networks regulated by multiple transcription factors. Gene ontology (GO) terms of development and morphogenesis are enriched in upregulated genes, while those of extracellular matrix and adhesion are enriched in downregulated genes. These results suggest that stable expression of a single gene Ngn1 in F3 cells induces not simply neurogenic but multifunctional changes that potentially affect the differentiation of human NSC via a reorganization of complex gene regulatory networks.

• Authors: Jun-ichi Satoh, Shinya Obayashi, Hiroko Tabunoki, Taeko Wakana, Seung U Kim
• Journal: Cellular and molecular neurobiology (Cell Mol Neurobiol) Vol. 30 Issue 3 Pg. 415-26 (Apr 2010) ISSN: 1573-6830 [Electronic] United States
• PMID: 19813087 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Basic Helix-Loop-Helix Transcription Factors
• Biomarkers
• LGR5 protein, human
• NEUROG1 protein, human
• Nerve Tissue Proteins
• Receptors, G-Protein-Coupled

Topics

• Basic Helix-Loop-Helix Transcription Factors (genetics)
• Biomarkers (metabolism)
• Cell Differentiation (genetics)
• Cell Line, Transformed
• Cell Lineage (genetics)
• Central Nervous System (embryology, growth & development, metabolism)
• Computational Biology
• Gene Expression Profiling
• Gene Expression Regulation (genetics)
• Gene Expression Regulation, Developmental (genetics)
• Gene Regulatory Networks (genetics)
• Genome-Wide Association Study
• Humans
• Molecular Biology (methods)
• Morphogenesis (genetics)
• Nerve Tissue Proteins (genetics)
• Neurons (cytology, metabolism)
• Receptors, G-Protein-Coupled (genetics)
• Software
• Stem Cells (cytology, metabolism)
• Transcriptional Activation (genetics)
• Up-Regulation (genetics)