MiR-34a targeting of notch ligand delta-like 1 impairs CD15 +/CD133 + tumor-propagating cells and supports neural differentiation in medulloblastoma

Pasqualino de Antonellis, Chiara Medaglia, Emilio Cusanelli, Immacolata Andolfo, Lucia Liguori, Gennaro de Vita, Marianeve Carotenuto, Annamaria Bello, Fabio Formiggini, Aldo Galeone, Giuseppe de Rosa, Antonella Virgilio, Immacolata Scognamiglio, Manuela Sciro, Giuseppe Basso, Johannes H. Schulte, Giuseppe Cinalli, Achille Iolascon, Massimo Zollo

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Background: Through negative regulation of gene expression, microRNAs (miRNAs) can function as oncosuppressors in cancers, and can themselves show altered expression in various tumor types. Here, we have investigated medulloblastoma tumors (MBs), which arise from an early impairment of developmental processes in the cerebellum, where Notch signaling is involved in many of the cell-fate-determining stages. Notch regulates a subset of MB cells that have stem-cell-like properties and can promote tumor growth. On the basis of this evidence, we hypothesized that miRNAs targeting the Notch pathway can regulate these phenomena, and can be used in anti-cancer therapies. Methodology/Principal Findings: In a screening of potential targets within Notch signaling, miR-34a was seen to be a regulator of the Notch pathway through its targeting of Notch ligand Delta-like 1 (Dll1). Down-regulation of Dll1 expression by miR-34a negatively regulates cell proliferation, and induces apoptosis and neural differentiation in MB cells. Using an inducible tetracycline on-off model of miR-34a expression, we show that in Daoy MB cells, Dll1 is the first target that is regulated in MB, as compared to the other targets analyzed here: Cyclin D1, cMyc and CDK4. MiR-34a expression negatively affects CD133 +/CD15 + tumor-propagating cells, then we assay through reverse-phase proteomic arrays, Akt and Stat3 signaling hypo-phosphorylation. Adenoviruses carrying the precursor miR-34a induce neurogenesis of tumor spheres derived from a genetic animal model of MB (Patch1 +/- p53 -/-), thus providing further evidence that the miR-34a/Dll1 axis controls both autonomous and non autonomous signaling of Notch. In vivo, miR-34a overexpression carried by adenoviruses reduces tumor burden in cerebellum xenografts of athymic mice, thus demonstrating an anti-tumorigenic role of miR-34a in vivo. Conclusions/Significance: Despite advances in our understanding of the pathogenesis of MB, one-third of patients with MB remain incurable. Here, we show that stable nucleic-acid-lipid particles carrying mature miR-34a can target Dll1 in vitro and show equal effects to those of adenovirus miR-34a cell infection. Thus, this technology forms the basis for their therapeutic use for the delivery of miR-34a in brain-tumor treatment, with no signs of toxicity described to date in non-human primate trials.

Original languageEnglish
Article numbere24584
JournalPLoS One
Volume6
Issue number9
DOIs
Publication statusPublished - Sep 12 2011

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Medulloblastoma
Tumors
Ligands
neoplasms
Neoplasms
Adenoviridae
MicroRNAs
cerebellum
Cerebellum
microRNA
cells
Phosphorylation
Somatostatin-Secreting Cells
Genetic Models
Cyclin D1
Neurogenesis
Cell proliferation
Gene Expression Regulation
therapeutics
Therapeutic Uses

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

MiR-34a targeting of notch ligand delta-like 1 impairs CD15 +/CD133 + tumor-propagating cells and supports neural differentiation in medulloblastoma. / de Antonellis, Pasqualino; Medaglia, Chiara; Cusanelli, Emilio; Andolfo, Immacolata; Liguori, Lucia; de Vita, Gennaro; Carotenuto, Marianeve; Bello, Annamaria; Formiggini, Fabio; Galeone, Aldo; de Rosa, Giuseppe; Virgilio, Antonella; Scognamiglio, Immacolata; Sciro, Manuela; Basso, Giuseppe; Schulte, Johannes H.; Cinalli, Giuseppe; Iolascon, Achille; Zollo, Massimo.

In: PLoS One, Vol. 6, No. 9, e24584, 12.09.2011.

Research output: Contribution to journalArticle

de Antonellis, P, Medaglia, C, Cusanelli, E, Andolfo, I, Liguori, L, de Vita, G, Carotenuto, M, Bello, A, Formiggini, F, Galeone, A, de Rosa, G, Virgilio, A, Scognamiglio, I, Sciro, M, Basso, G, Schulte, JH, Cinalli, G, Iolascon, A & Zollo, M 2011, 'MiR-34a targeting of notch ligand delta-like 1 impairs CD15 +/CD133 + tumor-propagating cells and supports neural differentiation in medulloblastoma', PLoS One, vol. 6, no. 9, e24584. https://doi.org/10.1371/journal.pone.0024584
de Antonellis, Pasqualino ; Medaglia, Chiara ; Cusanelli, Emilio ; Andolfo, Immacolata ; Liguori, Lucia ; de Vita, Gennaro ; Carotenuto, Marianeve ; Bello, Annamaria ; Formiggini, Fabio ; Galeone, Aldo ; de Rosa, Giuseppe ; Virgilio, Antonella ; Scognamiglio, Immacolata ; Sciro, Manuela ; Basso, Giuseppe ; Schulte, Johannes H. ; Cinalli, Giuseppe ; Iolascon, Achille ; Zollo, Massimo. / MiR-34a targeting of notch ligand delta-like 1 impairs CD15 +/CD133 + tumor-propagating cells and supports neural differentiation in medulloblastoma. In: PLoS One. 2011 ; Vol. 6, No. 9.
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T1 - MiR-34a targeting of notch ligand delta-like 1 impairs CD15 +/CD133 + tumor-propagating cells and supports neural differentiation in medulloblastoma

AU - de Antonellis, Pasqualino

AU - Medaglia, Chiara

AU - Cusanelli, Emilio

AU - Andolfo, Immacolata

AU - Liguori, Lucia

AU - de Vita, Gennaro

AU - Carotenuto, Marianeve

AU - Bello, Annamaria

AU - Formiggini, Fabio

AU - Galeone, Aldo

AU - de Rosa, Giuseppe

AU - Virgilio, Antonella

AU - Scognamiglio, Immacolata

AU - Sciro, Manuela

AU - Basso, Giuseppe

AU - Schulte, Johannes H.

AU - Cinalli, Giuseppe

AU - Iolascon, Achille

AU - Zollo, Massimo

PY - 2011/9/12

Y1 - 2011/9/12

N2 - Background: Through negative regulation of gene expression, microRNAs (miRNAs) can function as oncosuppressors in cancers, and can themselves show altered expression in various tumor types. Here, we have investigated medulloblastoma tumors (MBs), which arise from an early impairment of developmental processes in the cerebellum, where Notch signaling is involved in many of the cell-fate-determining stages. Notch regulates a subset of MB cells that have stem-cell-like properties and can promote tumor growth. On the basis of this evidence, we hypothesized that miRNAs targeting the Notch pathway can regulate these phenomena, and can be used in anti-cancer therapies. Methodology/Principal Findings: In a screening of potential targets within Notch signaling, miR-34a was seen to be a regulator of the Notch pathway through its targeting of Notch ligand Delta-like 1 (Dll1). Down-regulation of Dll1 expression by miR-34a negatively regulates cell proliferation, and induces apoptosis and neural differentiation in MB cells. Using an inducible tetracycline on-off model of miR-34a expression, we show that in Daoy MB cells, Dll1 is the first target that is regulated in MB, as compared to the other targets analyzed here: Cyclin D1, cMyc and CDK4. MiR-34a expression negatively affects CD133 +/CD15 + tumor-propagating cells, then we assay through reverse-phase proteomic arrays, Akt and Stat3 signaling hypo-phosphorylation. Adenoviruses carrying the precursor miR-34a induce neurogenesis of tumor spheres derived from a genetic animal model of MB (Patch1 +/- p53 -/-), thus providing further evidence that the miR-34a/Dll1 axis controls both autonomous and non autonomous signaling of Notch. In vivo, miR-34a overexpression carried by adenoviruses reduces tumor burden in cerebellum xenografts of athymic mice, thus demonstrating an anti-tumorigenic role of miR-34a in vivo. Conclusions/Significance: Despite advances in our understanding of the pathogenesis of MB, one-third of patients with MB remain incurable. Here, we show that stable nucleic-acid-lipid particles carrying mature miR-34a can target Dll1 in vitro and show equal effects to those of adenovirus miR-34a cell infection. Thus, this technology forms the basis for their therapeutic use for the delivery of miR-34a in brain-tumor treatment, with no signs of toxicity described to date in non-human primate trials.

AB - Background: Through negative regulation of gene expression, microRNAs (miRNAs) can function as oncosuppressors in cancers, and can themselves show altered expression in various tumor types. Here, we have investigated medulloblastoma tumors (MBs), which arise from an early impairment of developmental processes in the cerebellum, where Notch signaling is involved in many of the cell-fate-determining stages. Notch regulates a subset of MB cells that have stem-cell-like properties and can promote tumor growth. On the basis of this evidence, we hypothesized that miRNAs targeting the Notch pathway can regulate these phenomena, and can be used in anti-cancer therapies. Methodology/Principal Findings: In a screening of potential targets within Notch signaling, miR-34a was seen to be a regulator of the Notch pathway through its targeting of Notch ligand Delta-like 1 (Dll1). Down-regulation of Dll1 expression by miR-34a negatively regulates cell proliferation, and induces apoptosis and neural differentiation in MB cells. Using an inducible tetracycline on-off model of miR-34a expression, we show that in Daoy MB cells, Dll1 is the first target that is regulated in MB, as compared to the other targets analyzed here: Cyclin D1, cMyc and CDK4. MiR-34a expression negatively affects CD133 +/CD15 + tumor-propagating cells, then we assay through reverse-phase proteomic arrays, Akt and Stat3 signaling hypo-phosphorylation. Adenoviruses carrying the precursor miR-34a induce neurogenesis of tumor spheres derived from a genetic animal model of MB (Patch1 +/- p53 -/-), thus providing further evidence that the miR-34a/Dll1 axis controls both autonomous and non autonomous signaling of Notch. In vivo, miR-34a overexpression carried by adenoviruses reduces tumor burden in cerebellum xenografts of athymic mice, thus demonstrating an anti-tumorigenic role of miR-34a in vivo. Conclusions/Significance: Despite advances in our understanding of the pathogenesis of MB, one-third of patients with MB remain incurable. Here, we show that stable nucleic-acid-lipid particles carrying mature miR-34a can target Dll1 in vitro and show equal effects to those of adenovirus miR-34a cell infection. Thus, this technology forms the basis for their therapeutic use for the delivery of miR-34a in brain-tumor treatment, with no signs of toxicity described to date in non-human primate trials.

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