title: Shift of microRNA profile upon orthotopic xenografting of glioblastoma spheroid cultures.
abstract: Glioblastomas always recur despite surgery, radiotherapy and chemotherapy. A key player in the therapeutic resistance may be immature tumor cells with stem-like properties (TSCs) escaping conventional treatment. A group of promising molecular targets are microRNAs (miRs). miRs are small non-coding RNAs exerting post-transcriptional regulation of gene expression. In this study we aimed to identify over-expressed TSC-related miRs potentially amenable for therapeutic targeting. We used non-differentiated glioblastoma spheroid cultures (GSCs) containing TSCs and compared these to xenografts using a NanoString nCounter platform. This revealed 19 over-expressed miRs in the non-differentiated GSCs. Additionally, non-differentiated GSCs were compared to neural stem cells (NSCs) using a microarray platform. This revealed four significantly over-expressed miRs in the non-differentiated GSCs in comparison to the NSCs. The three most over-expressed miRs in the non-differentiated GSCs compared to xenografts were miR-126, -137 and -128. KEGG pathway analysis suggested the main biological function of these over-expressed miRs to be cell-cycle arrest and diminished proliferation. To functionally validate the profiling results suggesting association of these miRs with stem-like properties, experimental over-expression of miR-128 was performed. A consecutive limiting dilution assay confirmed a significantly elevated spheroid formation in the miR-128 over-expressing cells. This may provide potential therapeutic targets for anti-miRs to identify novel treatment options for GBM patients.
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 5000, Odense C, Denmark. bo.halle@rsyd.dk.|Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark. bo.halle@rsyd.dk.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark. bo.halle@rsyd.dk.
authors.forename: Bo
authors.initials: B
authors.lastname: Halle
authors.affiliation: Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.|Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Mads
authors.initials: M
authors.lastname: Thomassen
authors.affiliation: Regulus Therapeutics, 3545 John Hopkins Ct., Suite 210, San Diego, CA, 92121-1121, USA.
authors.forename: Ranga
authors.initials: R
authors.lastname: Venkatesan
authors.affiliation: Regulus Therapeutics, 3545 John Hopkins Ct., Suite 210, San Diego, CA, 92121-1121, USA.
authors.forename: Vivek
authors.initials: V
authors.lastname: Kaimal
authors.affiliation: Regulus Therapeutics, 3545 John Hopkins Ct., Suite 210, San Diego, CA, 92121-1121, USA.
authors.forename: Eric G
authors.initials: EG
authors.lastname: Marcusson
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 5000, Odense C, Denmark.|Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.
authors.forename: Sune
authors.initials: S
authors.lastname: Munthe
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.
authors.forename: Mia D
authors.initials: MD
authors.lastname: Sørensen
authors.affiliation: Department of Nuclear Medicine, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Aaberg-Jessen
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.
authors.forename: Stine S
authors.initials: SS
authors.lastname: Jensen
authors.affiliation: Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 21, 5000, Odense C, Denmark.
authors.forename: Morten
authors.initials: M
authors.lastname: Meyer
authors.affiliation: Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.|Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Torben A
authors.initials: TA
authors.lastname: Kruse
authors.affiliation: Faculty of Health Sciences, Lundbeckfonden Center of Excellence NanoCAN and Molecular Oncology, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 25, 5000, Odense C, Denmark.
authors.forename: Helle
authors.initials: H
authors.lastname: Christiansen
authors.affiliation: Faculty of Health Sciences, Lundbeckfonden Center of Excellence NanoCAN and Molecular Oncology, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 25, 5000, Odense C, Denmark.
authors.forename: Steffen
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Faculty of Health Sciences, Lundbeckfonden Center of Excellence NanoCAN and Molecular Oncology, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 25, 5000, Odense C, Denmark.
authors.forename: Jan
authors.initials: J
authors.lastname: Mollenhauer
authors.affiliation: Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.
authors.forename: Mette K
authors.initials: MK
authors.lastname: Schulz
authors.affiliation: Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.
authors.forename: Claus
authors.initials: C
authors.lastname: Andersen
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark.
authors.forename: Bjarne W
authors.initials: BW
authors.lastname: Kristensen
mesh_terms: D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic; D005909:Glioblastoma / Q000378:metabolism* / Q000473:pathology; D064593:Heterografts; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D046228:Microarray Analysis; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D058953:Neural Stem Cells / Q000378:metabolism; D011923:Rats, Nude; D018874:Spheroids, Cellular / Q000637:transplantation
chemical_list: D035683:MicroRNAs
keywords: Cancer stem cell; Glioblastoma; MicroRNA; Tumor stem cell