Medullospheres from DAOY, UW228 and ONS-76 Cells: Increased Stem Cell Population and Proteomic Modifications

Zanini, Cristina; Ercole, Elisabetta; Mandili, Giorgia; Salaroli, Roberta; Poli, Alice; Renna, Cristiano; Papa, Valentina; Cenacchi, Giovanna; Forni, Marco

doi:10.1371/journal.pone.0063748

Cited by 36 publications

(42 citation statements)

References 36 publications

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“…2B). This indicates that medulloblastoma cells grown as neurospheres adopt CSC characteristics, which is consistent with previous reports (35). Additionally, neuro-spheres depicted substantial increase in phosphorylation of AKT on Ser-473, indicative of activation of the PI3K/AKT pathway (Fig.…”

Section: Resultssupporting

confidence: 91%

Potent Antineoplastic Effects of Combined PI3Kα–MNK Inhibition in Medulloblastoma

Eckerdt

Bell

Beauchamp

et al. 2019

Molecular Cancer Research

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Medulloblastoma is a highly malignant pediatric brain tumor associated with poor outcome. Developing treatments that target the cancer stem cell (CSC) population in medulloblastoma are important to prevent tumor relapse and induce long-lasting clinical responses. We utilized medulloblastoma neurospheres that display CSC characteristics and found activation of the PI3K/AKT pathway in sphere-forming cells. Of all class I A PI3Ks, only the PI3Ka isoform was required for sphere formation by medulloblastoma cells. Knockdown of p110a, but not p110b or p110d, significantly disrupted cancer stem cell frequencies as determined by extreme limiting dilution analysis (ELDA), indicating an essential role for the PI3Ka catalytic isoform in medulloblastoma CSCs. Importantly, pharmacologic inhibition of the MAPK-interacting kinase (MNK) enhanced the antineoplastic effects of targeted PI3Ka inhibition in medulloblastoma. This indicates that MNK signaling promotes survival in medulloblastoma, suggesting dual PI3Ka and MNK inhibition may provide a novel approach to target and eliminate medulloblastoma CSCs. We also observed a significant reduction in tumor formation in subcutaneous and intracranial mouse xenograft models, which further suggests that this combinatorial approach may represent an efficient therapeutic strategy for medulloblastoma.Implications: These findings raise the possibility of a unique therapeutic approach for medulloblastoma, involving MNK targeting to sensitize medulloblastoma CSCs to PI3Ka inhibition.

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Section: Resultssupporting

confidence: 91%

Potent Antineoplastic Effects of Combined PI3Kα–MNK Inhibition in Medulloblastoma

Eckerdt

Bell

Beauchamp

et al. 2019

Molecular Cancer Research

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“…Although Daoy and UW228 are commonly used in studies of medulloblastoma, very few proteomic studies related to Daoy and UW228 have been published [28,30], with almost no studies performed by integrated transcriptomic and proteomic analysis. Given the distinct developmental origins of the MB subtypes, driving the heterogeneity within and between the different subtypes, identification and validation of predictive and prognostic signature biomarkers for each subtype, with utility for diagnosis, management and follow-up of the disease may ultimately improve survival.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, Martelli et al [29] performed a top-down LC-MS proteomic analysis of Daoy, which identified 53 proteins. Conversely, the only detailed proteomics study for UW228 was reported by Zanini et al [30], who compared the proteomes of Daoy and UW228, cultured in adherence or as medullospheres, to determine the influence of growth conditions on the modulation of protein expression. Moreover, the exact MB subtype to which Daoy and UW228 belong is not clearly established.…”

Section: Introductionmentioning

confidence: 99%

Integrated Proteomic and Transcriptomic-Based Approaches to Identifying Signature Biomarkers and Pathways for Elucidation of Daoy and UW228 Subtypes

et al. 2017

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Medulloblastoma (MB) is the most common malignant pediatric brain tumor. Patient survival has remained largely the same for the past 20 years, with therapies causing significant health, cognitive, behavioral and developmental complications for those who survive the tumor. In this study, we profiled the total transcriptome and proteome of two established MB cell lines, Daoy and UW228, using high-throughput RNA sequencing (RNA-Seq) and label-free nano-LC-MS/MS-based quantitative proteomics, coupled with advanced pathway analysis. While Daoy has been suggested to belong to the sonic hedgehog (SHH) subtype, the exact UW228 subtype is not yet clearly established. Thus, a goal of this study was to identify protein markers and pathways that would help elucidate their subtype classification. A number of differentially expressed genes and proteins, including a number of adhesion, cytoskeletal and signaling molecules, were observed between the two cell lines. While several cancer-associated genes/proteins exhibited similar expression across the two cell lines, upregulation of a number of signature proteins and enrichment of key components of SHH and WNT signaling pathways were uniquely observed in Daoy and UW228, respectively. The novel information on differentially expressed genes/proteins and enriched pathways provide insights into the biology of MB, which could help elucidate their subtype classification.

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“…Neurosphere formation was used as an established experimental assay to evaluate cancer stem cell proliferation (Nör et al, 2013;Zanini et al, 2013). Daoy and D283 cells were dissociated with trypsin-EDTA into a single cell suspension and seeded at 500 cells/well in 24-well low-attachment plates.…”

Section: Neurosphere Formationmentioning

confidence: 99%

HDAC and MAPK/ERK Inhibitors Cooperate to Reduce Viability and Stemness in Medulloblastoma

Jaeger

Ghisleni

Cardoso

et al. 2019

Preprint

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Medulloblastoma (MB), which originates from embryonic neural stem cells (NSCs) or neural precursors in the developing cerebellum, is the most common malignant brain tumor of childhood.Recurrent and metastatic disease is the principal cause of death and may be related to resistance within cancer stem cells (CSCs). Chromatin state is involved in maintaining signaling pathways related to stemness, and inhibition of histone deacetylase enzymes (HDAC) has emerged as a therapeutic tool to target this cell population. Here, we observed changes in the stemness phenotype induced by HDAC inhibition in MB. Analyses of MB tumor samples showed that the stemness markers BMI1 and CD133 are expressed in all molecular subgroups of MB. We then treated human MB cells with the HDAC inhibitor (HDACi) sodium butyrate (NaB). Increased acetylation mediated by NaB reduced cell viability and expression of BMI1 and CD133. Activation of extracellular-regulated kinase (ERK) signaling was also decreased after treatment with NaB.Enrichment analysis of genes associated with CD133 or BMI1 expression showed mitogenactivated protein kinase (MAPK)/ERK signaling and related pathways as the most enriched processes. We went on to use the mitogen-activated protein kinase kinase (MEK) inhibitor U0126 to explore the role of MAPK/ERK pathway. MEK inhibition reduced expression of the stemness markers, hindered MB neurosphere formation, and its antiproliferative effect was enhanced by combination with NaB. These results suggest that combining HDAC and MAPK/ERK inhibitors may be more effective in reducing MB proliferation than single-drug treatments, through modulation of the stemness phenotype of MB cells. 5 Keywords: Histone deacetylase Extracellular-regulated kinase Stemness Medulloblastoma Brain tumor 6

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Medullospheres from DAOY, UW228 and ONS-76 Cells: Increased Stem Cell Population and Proteomic Modifications

Cited by 36 publications

References 36 publications

Potent Antineoplastic Effects of Combined PI3Kα–MNK Inhibition in Medulloblastoma

Potent Antineoplastic Effects of Combined PI3Kα–MNK Inhibition in Medulloblastoma

Integrated Proteomic and Transcriptomic-Based Approaches to Identifying Signature Biomarkers and Pathways for Elucidation of Daoy and UW228 Subtypes

HDAC and MAPK/ERK Inhibitors Cooperate to Reduce Viability and Stemness in Medulloblastoma

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