Long-term Cultured Human Neural Stem Cells Undergo Spontaneous Transformation to Tumor-Initiating Cells

In this report, we describe the spontaneous malignant transformation of long-term cultured human fetal striatum neural stem cells (hsNSCs, passage 17). After subcutaneous transplantation of long-term cultured hsNSCs into immunodeficient nude mice, 2 out of 15 mice formed xenografts which expressed neuroendocrine tumor markers CgA and NSE. T1 cells, a cell line that we derived from one of the two subcutaneous xenografts, have undergone continuous expansion in vitro. These T1 cells showed stem cell-like features and expressed neural stem cell markers nestin and CD133. The T1 cells were involved in abnormal karyotype, genomic instability and fast proliferation. Importantly, after long-term in vitro culture, the T1 cells did not result in subcutaneous xenografts, but induced intracranial tumor formation, indicating that they adjusted themselves to the intracranial microenvironment. We further found that the T1 cells exhibited an overexpressed level of EGFR, and the CD133 positive T1 cells showed a truncation mutation in the exons 2-7 of the EGFR (EGFRvIII) gene. These results suggest that continuous expansion of neural stem cells in culture may lead to malignant spontaneous transformation. This phenomenon may be functionally related to EGFR by EGFRvIII gene mutation.

[1]  L. Ricci-Vitiani,et al.  Tumorigenic Potential of Olfactory Bulb-Derived Human Adult Neural Stem Cells Associates with Activation of TERT and NOTCH1 , 2009, PloS one.

[2]  Kouji Matsushima,et al.  The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract , 1998, Nature.

[3]  L. Chin,et al.  Malignant astrocytic glioma: genetics, biology, and paths to treatment. , 2007, Genes & development.

[4]  W. Cavenee,et al.  EGFR gene amplification ‐ rearrangement in human glioblastomas , 1995, International journal of cancer.

[5]  B. van Deurs,et al.  EGFRvIII escapes down-regulation due to impaired internalization and sorting to lysosomes. , 2007, Carcinogenesis.

[6]  J. Schlessinger,et al.  Signaling by Receptor Tyrosine Kinases , 1993 .

[7]  S. Schulz,et al.  Enhanced expression of the CXCl12/SDF-1 chemokine receptor CXCR7 after cerebral ischemia in the rat brain , 2008, Journal of Neuroimmunology.

[8]  R. McKay,et al.  CNS stem cells express a new class of intermediate filament protein , 1990, Cell.

[9]  I. Weissman,et al.  Transplanted human fetal neural stem cells survive, migrate, and differentiate in ischemic rat cerebral cortex. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Yoshitaka Narita,et al.  Tumor heterogeneity is an active process maintained by a mutant EGFR-induced cytokine circuit in glioblastoma. , 2010, Genes & development.

[11]  R. Salgia,et al.  Chemokine Receptors CXCR-1/2 Activate Mitogen-activated Protein Kinase via the Epidermal Growth Factor Receptor in Ovarian Cancer Cells* , 2000, The Journal of Biological Chemistry.

[12]  G. Schettini,et al.  CXCR4 Activation Induces Epidermal Growth Factor Receptor Transactivation in an Ovarian Cancer Cell Line , 2004, Annals of the New York Academy of Sciences.

[13]  Qiuyue Du,et al.  Correlation effect of EGFR and CXCR4 and CCR7 chemokine receptors in predicting breast cancer metastasis and prognosis , 2010, Journal of experimental & clinical cancer research : CR.

[14]  B. Liu,et al.  Hypoxia-inducible factor (HIF)-1 alpha directly enhances the transcriptional activity of stem cell factor (SCF) in response to hypoxia and epidermal growth factor (EGF). , 2008, Carcinogenesis.

[15]  E. Lagasse,et al.  Cancer stem cells with genetic instability: the best vehicle with the best engine for cancer , 2008, Gene Therapy.

[16]  S. Morrison,et al.  Prospective identification of tumorigenic breast cancer cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[17]  S. Gabriel,et al.  Epidermal Growth Factor Receptor Activation in Glioblastoma through Novel Missense Mutations in the Extracellular Domain , 2006, PLoS medicine.

[18]  Daniel H. Geschwind,et al.  Cancerous stem cells can arise from pediatric brain tumors , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Jayant P. Menon,et al.  Neuronal and glioma-derived stem cell factor induces angiogenesis within the brain. , 2006, Cancer cell.

[20]  C. Brinckerhoff,et al.  CXCR4 and Matrix Metalloproteinase-1 Are Elevated in Breast Carcinoma–Associated Fibroblasts and in Normal Mammary Fibroblasts Exposed to Factors Secreted by Breast Cancer Cells , 2009, Molecular Cancer Research.

[21]  George M. Smith,et al.  Targeting Sensory Axon Regeneration in Adult Spinal Cord , 2007, The Journal of Neuroscience.

[22]  J. Schlessinger.  Cell Signaling by Receptor Tyrosine Kinases , 2000, Cell.

[23]  T. Kipps,et al.  CXCR4: a key receptor in the crosstalk between tumor cells and their microenvironment. , 2006, Blood.

[24]  R. DePinho,et al.  Malignant glioma: genetics and biology of a grave matter. , 2001, Genes & development.

[25]  Paul Workman,et al.  EGFRvIII Deletion Mutations in Pediatric High-Grade Glioma and Response to Targeted Therapy in Pediatric Glioma Cell Lines , 2009, Clinical Cancer Research.

[26]  U. Lendahl,et al.  Generalized potential of adult neural stem cells. , 2000, Science.

[27]  J. Schlessinger,et al.  Cell Signaling by Receptor Tyrosine Kinases , 2000, Cell.

[28]  I. Bayazitov,et al.  A perivascular niche for brain tumor stem cells. , 2007, Cancer cell.

[29]  R. Gilbertson,et al.  Making a tumour's bed: glioblastoma stem cells and the vascular niche , 2007, Nature Reviews Cancer.

[30]  F. Gage,et al.  Regenerating the damaged central nervous system , 2000, Nature.

[31]  M. Moran,et al.  Epidermal growth factor receptor variant III-induced glioma invasion is mediated through myristoylated alanine-rich protein kinase C substrate overexpression. , 2009, Cancer research.

[32]  P. Vogt,et al.  In vivo transformation of neural stem cells following transplantation in the injured nervous system. , 2010, Journal of reconstructive microsurgery.

[33]  Samia J. Khoury,et al.  Directed migration of neural stem cells to sites of CNS injury by the stromal cell-derived factor 1α/CXC chemokine receptor 4 pathway , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[34]  R. Thompson,et al.  Stem cells as vehicles for the treatment of brain cancer. , 2007, Neurosurgery clinics of North America.

[35]  S. Sorscher.  EGFR mutations and sensitivity to gefitinib. , 2004, The New England journal of medicine.

[36]  Li Shen,et al.  Isolation and characterization of neural stem cells from human fetal striatum. , 2005, Biochemical and biophysical research communications.

[37]  I. Blümcke,et al.  Spontaneous In Vitro Transformation of Adult Neural Precursors into Stem‐Like Cancer Cells , 2009, Brain pathology.

[38]  P. Lønning,et al.  Long-term cultures of bone marrow-derived human mesenchymal stem cells frequently undergo spontaneous malignant transformation. , 2009, Cancer research.

[39]  Angelo L. Vescovi,et al.  Brain tumour stem cells , 2006, Nature Reviews Cancer.

[40]  R. Nicholson,et al.  EGFR and cancer prognosis. , 2001, European journal of cancer.

[41]  A. Björklund,et al.  Cell replacement therapies for central nervous system disorders , 2000, Nature Neuroscience.

[42]  Gideon Rechavi,et al.  Donor-Derived Brain Tumor Following Neural Stem Cell Transplantation in an Ataxia Telangiectasia Patient , 2009, PLoS medicine.