DIPG-25. IN VITRO AND IN VIVO ANALYSIS OF TENASCIN-C EXPRESSION IN PEDIATRIC BRAINSTEM GLIOMA
| Autor: | Tadanori Tomita, C. David James, Tina Huang, Jin Qi, Rintaro Hashizume, Amanda Saratsis |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Cancer Research
biology business.industry Tenascin C In vivo analysis medicine.disease musculoskeletal system Pediatric Brainstem Glioma In vitro Abstracts Oncology Glioma Gene expression Cancer research biology.protein Medicine Immunohistochemistry Neurology (clinical) business Protein overexpression |
| Popis: | INTRODUCTION: Diffuse intrinsic pontine glioma (DIPG) is a highly morbid pediatric brainstem tumor. Tenascin-C (TNC) is an extracellular matrix protein associated with NOTCH pathway activation, and is expressed during normal brain development by oligodendroglial cells (OPCs), the purported DIPG cell of origin. We previously reported TNC overexpression in DIPG tissue and CSF, associated with Histone H3K27M mutation. We therefore investigated the pattern and effect of TNC expression using pediatric glioma primary cell lines. METHODS: To determine the molecular effects of altered TNC expression in DIPG in vitro, we evaluated endogenous TNC expression in DIPG cell lines (n=3) and compared to TNC suppression and over-expression via vector-mediated shRNA knockdown and cDNA amplification, respectively. Effects of genetic manipulations were confirmed by western blot and qPCR. Cell proliferation, migration and adhesion were compared, as well as patterns of gene expression (RNA-Seq) across cell lines. Luciferase-expressing DIPG cells modified with TNC shRNA were used to create mouse intracranial xenografts. Animal survival and tumor growth was compared between groups, and TNC expression confirmed by immunohistochemistry. RESULTS: shRNA knockdown achieved up to 80% reduction in TNC expression, inhibiting cell proliferation and migration on functional analysis compared to control DIPG lines. Distinct pathways of gene expression favoring dedifferentiation and stem-like state correlated with higher TNC expression. Anaylsis cDNA and in vivo studies is currently underway. CONCLUSIONS: Here, we report the pattern and effect of modulated TNC expression in pediatric brainstem glioma (DIPG). Our findings suggest TNC may play a role in cell proliferation, migration, and maintaining stem-ness, possibly contributing to brainstem gliomagenesis. Further in vitro and in vivo studies exploring the mechanism of overexpression and effects of targeting TNC expression in DIPG are planned. |
| Databáze: | OpenAIRE |
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