Related Experiment Video
Updated: Nov 9, 2025
Mosaic Zebrafish Transgenesis for Functional Genomic Analysis of Candidate Cooperative Genes in Tumor Pathogenesis
Published on: March 31, 2015
Combining inhibitors of Brd4 and cyclin-dependent kinase can decrease tumor growth in neuroblastoma with MYCN
Lauren Wood1, Min Huang2, Jasmine Zeki1
1Department of Surgery, Stanford University, Stanford, CA, USA.
Introduction:
High-risk neuroblastoma is a deadly disease; poor prognosticators are MYCN-amplification and TERT-overexpression. We hypothesized that Gene Set Enrichment Analysis (GSEA) could identify pathways associated with MYCN-amplification and that inhibition of these pathways could decrease tumor growth.
Methods:
We analyzed the Neuroblastoma-Kocak dataset (GSE45547, n = 649) and identified pathways associated with MYCN-amplification. Inhibitors were selected from upregulated gene sets for in vitro cytotoxicity testing using ST16-patient-derived primary neuroblastoma cells and in vivo testing using orthotopic ST16-patient-derived xenografts (PDX) in mice. Tumor volume was measured with ultrasound and tumor sections examined after H&E staining.
Results:
GSEA identified significantly overexpressed gene sets in MYCN-amplified tumors including MYC targets, cell cycle mitotic genes, TERT associated genes, loss of RB1 gene sets, and E2Fs targets. Several genes were potential Bromodomain-containing protein 4 (Brd4) targets, making Brd4 inhibitors - JQ1, AZD5153 - and cyclin-dependent kinase (Brd4's binding partner) inhibitors - dinaciclib - potential therapeutic agents. JQ1 and dinaciclib were synergistic in inducing cytotoxicity in vitro. Dinaciclib-AZD5153 in vivo decreased tumor size compared to control, and increased tumor lymphocyte infiltration and necrosis on histology.
Conclusions:
GSEA is a powerful approach to identify upregulated genes and potential therapeutic targets. Dinaciclib-AZD5153 combination therapy can be effective against MYCN-amplified and TERT-overexpressing neuroblastoma tumors.
Insights
Gene Set Enrichment Analysis identified therapeutic targets for high-risk neuroblastoma. Combination therapy with dinaciclib and AZD5153 effectively reduced tumor growth in preclinical models.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- High-risk neuroblastoma presents a significant clinical challenge with poor prognoses often linked to MYCN-amplification and TERT-overexpression.
- Gene Set Enrichment Analysis (GSEA) offers a potential method to uncover biological pathways associated with these poor prognostic markers.
Purpose of the Study:
- To identify pathways dysregulated in MYCN-amplified neuroblastoma using GSEA.
- To evaluate the therapeutic potential of inhibiting identified pathways in preclinical models of neuroblastoma.
Main Methods:
- GSEA was performed on the Neuroblastoma-Kocak dataset (GSE45547) to identify gene sets associated with MYCN-amplification.
- In vitro cytotoxicity assays and in vivo orthotopic xenograft models (PDX) were used to test selected inhibitors, including Brd4 inhibitors (JQ1, AZD5153) and a CDK inhibitor (dinaciclib).
- Tumor growth was monitored via ultrasound, and histological analysis (H&E staining) assessed treatment effects.
Main Results:
- GSEA revealed significant enrichment of MYC targets, cell cycle genes, TERT-associated genes, RB1 loss, and E2F targets in MYCN-amplified tumors.
- Bromodomain-containing protein 4 (Brd4) and cyclin-dependent kinase (CDK) pathways emerged as potential therapeutic targets.
- Combination therapy with dinaciclib and AZD5153 demonstrated synergistic cytotoxicity in vitro and significantly reduced tumor volume, increased lymphocyte infiltration, and induced necrosis in vivo.
Conclusions:
- GSEA is a valuable tool for identifying actionable molecular targets in complex cancers like neuroblastoma.
- The combination of dinaciclib and AZD5153 shows promise as an effective therapeutic strategy for MYCN-amplified and TERT-overexpressing neuroblastoma.
Related Concept Videos
02:34Inhibition of Cdk Activity
02:50Combination Therapies and Personalized Medicine
The combination of the drug acetazolamide and sulforaphane is a good example of combination therapy to treat cancer. The cells in the interior of a large tumor often die due to the hypoxic and...
02:57Targeted Cancer Therapies
There are several types of targeted therapies against...
02:38Mitogens and the Cell Cycle
02:23Abnormal Proliferation
02:15M-Cdk Drives Transition Into Mitosis
Cyclin-dependent kinases, or Cdks, work in concert with cyclins to control cell cycle transitions. M-Cdk, a complex of Cdk1 bound to M cyclin, is a well-known example of this coordinated control that drives the transition from the G2 to the M phase.
M cyclin...