title: ST3GAL1-Associated Transcriptomic Program in Glioblastoma Tumor Growth, Invasion, and Prognosis.
abstract: BACKGROUND
  Cell surface sialylation is associated with tumor cell invasiveness in many cancers. Glioblastoma is the most malignant primary brain tumor and is highly infiltrative. ST3GAL1 sialyltransferase gene is amplified in a subclass of glioblastomas, and its role in tumor cell self-renewal remains unexplored.
  
  
  METHODS
  Self-renewal of patient glioma cells was evaluated using clonogenic, viability, and invasiveness assays. ST3GAL1 was identified from differentially expressed genes in Peanut Agglutinin-stained cells and validated in REMBRANDT (n = 390) and Gravendeel (n = 276) clinical databases. Gene set enrichment analysis revealed upstream processes. TGFβ signaling on ST3GAL1 transcription was assessed using chromatin immunoprecipitation. Transcriptome analysis of ST3GAL1 knockdown cells was done to identify downstream pathways. A constitutively active FoxM1 mutant lacking critical anaphase-promoting complex/cyclosome ([APC/C]-Cdh1) binding sites was used to evaluate ST3Gal1-mediated regulation of FoxM1 protein. Finally, the prognostic role of ST3Gal1 was determined using an orthotopic xenograft model (3 mice groups comprising nontargeting and 2 clones of ST3GAL1 knockdown in NNI-11 [8 per group] and NNI-21 [6 per group]), and the correlation with patient clinical information. All statistical tests on patients' data were two-sided; other P values below are one-sided.
  
  
  RESULTS
  High ST3GAL1 expression defines an invasive subfraction with self-renewal capacity; its loss of function prolongs survival in a mouse model established from mesenchymal NNI-11 (P < .001; groups of 8 in 3 arms: nontargeting, C1, and C2 clones of ST3GAL1 knockdown). ST3GAL1 transcriptomic program stratifies patient survival (hazard ratio [HR] = 2.47, 95% confidence interval [CI] = 1.72 to 3.55, REMBRANDT P = 1.92 x 10⁻⁸; HR = 2.89, 95% CI = 1.94 to 4.30, Gravendeel P = 1.05 x 10⁻¹¹), independent of age and histology, and associates with higher tumor grade and T2 volume (P = 1.46 x 10⁻⁴). TGFβ signaling, elevated in mesenchymal patients, correlates with high ST3GAL1 (REMBRANDT gliomacor = 0.31, P = 2.29 x 10⁻¹⁰; Gravendeel gliomacor = 0.50, P = 3.63 x 10⁻²⁰). The transcriptomic program upon ST3GAL1 knockdown enriches for mitotic cell cycle processes. FoxM1 was identified as a statistically significantly modulated gene (P = 2.25 x 10⁻⁵) and mediates ST3Gal1 signaling via the (APC/C)-Cdh1 complex.
  
  
  CONCLUSIONS
  The ST3GAL1-associated transcriptomic program portends poor prognosis in glioma patients and enriches for higher tumor grades of the mesenchymal molecular classification. We show that ST3Gal1-regulated self-renewal traits are crucial to the sustenance of glioblastoma multiforme growth.
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Yuk Kien
authors.initials: YK
authors.lastname: Chong
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Edwin
authors.initials: E
authors.lastname: Sandanaraj
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Lynnette W H
authors.initials: LW
authors.lastname: Koh
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Moogaambikai
authors.initials: M
authors.lastname: Thangaveloo
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Melanie S Y
authors.initials: MS
authors.lastname: Tan
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Geraldene R H
authors.initials: GR
authors.lastname: Koh
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Tan Boon
authors.initials: TB
authors.lastname: Toh
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Grace G Y
authors.initials: GG
authors.lastname: Lim
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Joanna D
authors.initials: JD
authors.lastname: Holbrook
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Oi Lian
authors.initials: OL
authors.lastname: Kon
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Mahendran
authors.initials: M
authors.lastname: Nadarajah
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Ivan
authors.initials: I
authors.lastname: Ng
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Wai Hoe
authors.initials: WH
authors.lastname: Ng
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Nguan Soon
authors.initials: NS
authors.lastname: Tan
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA).
authors.forename: Kah Leong
authors.initials: KL
authors.lastname: Lim
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA). carol_tang@nni.com.sg beng_ti_ang@nni.com.sg yuk_kien_chong@nni.com.sg.
authors.forename: Carol
authors.initials: C
authors.lastname: Tang
authors.affiliation: Department of Research (YKC, ES, LWHK, MT, MSYT, GRHK, TBT, GGYL, KLL, CT), Department of Neuroradiology (MN), and Department of Neurosurgery (IN, WHN, BTA), National Neuroscience Institute, Singapore; Department of Physiology (YKC, KLL, BTA) and Department of Biochemistry (OLK), Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Singapore Institute for Clinical Sciences (ES, JDH, BTA) and Institute of Molecular and Cell Biology (NST), Agency for Science, Technology and Research (A*STAR), Singapore; School of Biological Sciences, Nanyang Technological University, Singapore (ES, LWHK, MSYT, NST); Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore (OLK, CT); Duke-National University of Singapore Graduate Medical School, Singapore (IN, WHN, KLL, CT, BTA). carol_tang@nni.com.sg beng_ti_ang@nni.com.sg yuk_kien_chong@nni.com.sg.
authors.forename: Beng Ti
authors.initials: BT
authors.lastname: Ang
mesh_terms: D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology*; D049109:Cell Proliferation; D002470:Cell Survival; D047369:Chromatin Immunoprecipitation; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D005909:Glioblastoma / Q000378:metabolism* / Q000473:pathology*; D064593:Heterografts; D006801:Humans; D053208:Kaplan-Meier Estimate; D051379:Mice; D009361:Neoplasm Invasiveness; D011379:Prognosis; D012799:Sialyltransferases / Q000378:metabolism*; D015398:Signal Transduction; D059467:Transcriptome; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014410:Tumor Stem Cell Assay; D015854:Up-Regulation; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase
chemical_list: C107585:FOXM1 protein, human; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors; C491524:Foxm1 protein, mouse; D016212:Transforming Growth Factor beta; D012799:Sialyltransferases; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase