Gamble, LD, Purgato, S, Murray, J, Xiao, L, Yu, DMT, Hanssen, KM, Giorgi, FM, Carter, DR, Gifford, AJ, Valli, E, Milazzo, G, Kamili, A, Mayoh, C, Liu, B, Eden, G, Sarraf, S, Allan, S, Giacomo, SD, Flemming, CL, Russell, AJ, Cheung, BB, Oberthuer, A, London, WB, Fischer, M, Trahair, TN, Fletcher, JI, Marshall, GM, Ziegler, DS, Hogarty, MD, Burns, MR, Perini, G, Norris, MD & Haber, M 2019, 'Inhibition of polyamine synthesis and uptake reduces tumor progression and prolongs survival in mouse models of neuroblastoma', Science Translational Medicine, vol. 11, no. 477.View/Download from: UTS OPUS or Publisher's site
Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Amplification of the MYCN oncogene is associated with an aggressive phenotype and poor outcome in childhood neuroblastoma. Polyamines are highly regulated essential cations that are frequently elevated in cancer cells, and the rate-limiting enzyme in polyamine synthesis, ornithine decarboxylase 1 (ODC1), is a direct transcriptional target of MYCN. Treatment of neuroblastoma cells with the ODC1 inhibitor difluoromethylornithine (DFMO), although a promising therapeutic strategy, is only partially effective at impeding neuroblastoma cell growth due to activation of compensatory mechanisms resulting in increased polyamine uptake from the surrounding microenvironment. In this study, we identified solute carrier family 3 member 2 (SLC3A2) as the key transporter involved in polyamine uptake in neuroblastoma. Knockdown of SLC3A2 in neuroblastoma cells reduced the uptake of the radiolabeled polyamine spermidine, and DFMO treatment increased SLC3A2 protein. In addition, MYCN directly increased polyamine synthesis and promoted neuroblastoma cell proliferation by regulating SLC3A2 and other regulatory components of the polyamine pathway. Inhibiting polyamine uptake with the small-molecule drug AMXT 1501, in combination with DFMO, prevented or delayed tumor development in neuroblastoma-prone mice and extended survival in rodent models of established tumors. Our findings suggest that combining AMXT 1501 and DFMO with standard chemotherapy might be an effective strategy for treating neuroblastoma.
Dorneburg, C, Fischer, M, Barth, TFE, Mueller-Klieser, W, Hero, B, Gecht, J, Carter, DR, de Preter, K, Mayer, B, Christner, L, Speleman, F, Marshall, GM, Debatin, K-M & Beltinger, C 2018, 'LDHA in Neuroblastoma Is Associated with Poor Outcome and Its Depletion Decreases Neuroblastoma Growth Independent of Aerobic Glycolysis.', Clinical cancer research : an official journal of the American Association for Cancer Research, vol. 24, no. 22, pp. 5772-5783.View/Download from: UTS OPUS or Publisher's site
Purpose: To investigate whether lactate dehydrogenase A (LDHA), an important component of the LDH tetramer crucial for aerobic glycolysis, is associated with patient outcome and constitutes a therapeutic target in neuroblastoma (NB).Experimental Design: Expression of LDHA mRNA and protein was determined in 709 and 110 NB patient samples, respectively, and correlated with survival and risk factors. LDHA and LDHB were depleted in human NB cell lines by CRISPR/Cas9 and shRNA, respectively, and aerobic glycolysis, clonogenicity, and tumorigenicity were determined. Expression of LDHA in relation to MYCN was measured in NB cell lines and in the TH-MYCN NB mouse model.Results: Expression of LDHA, both on the mRNA and the protein level, was significantly and independently associated with decreased patient survival. Predominant cytoplasmic localization of LDHA protein was associated with poor outcome. Amplification and expression of MYCN did not correlate with expression of LDHA in NB cell lines or TH-MYCN mice, respectively. Knockout of LDHA inhibited clonogenicity, tumorigenicity, and tumor growth without abolishing LDH activity or significantly decreasing aerobic glycolysis. Concomitant depletion of LDHA and the isoform LDHB ablated clonogenicity while not abrogating LDH activity or decreasing aerobic glycolysis. The isoform LDHC was not expressed.Conclusions: High expression of LDHA is independently associated with outcome of NB, and NB cells can be inhibited by depletion of LDHA or LDHB. This inhibition appears to be unrelated to LDH activity and aerobic glycolysis. Thus, investigations of inhibitory mechanisms beyond attenuation of aerobic glycolysis are warranted, both in NB and normal cells. Clin Cancer Res; 24(22); 5772-83. ©2018 AACR.
Ooi, CY, Carter, DR, Liu, B, Mayoh, C, Beckers, A, Lalwani, A, Nagy, Z, De Brouwer, S, Decaesteker, B, Hung, TT, Norris, MD, Haber, M, Liu, T, De Preter, K, Speleman, F, Cheung, BB & Marshall, GM 2018, 'Network modeling of microRNA-mRNA interactions in neuroblastoma tumorigenesis identifies miR-204 as a direct inhibitor of MYCN', Cancer Research, vol. 78, no. 12, pp. 3122-3134.View/Download from: UTS OPUS or Publisher's site
© 2018 American Association for Cancer Research. Neuroblastoma is a pediatric cancer of the sympathetic nervous system where MYCN amplification is a key indicator of poor prognosis. However, mechanisms by which MYCN promotes neuroblastoma tumorigenesis are not fully understood. In this study, we analyzed global miRNA and mRNA expression profiles of tissues at different stages of tumorigenesis from TH-MYCN transgenic mice, a model of MYCN-driven neuroblastoma.Onthe basis of a Bayesian learning network model in which wecompared pretumor ganglia from TH-MYCN+/+ mice to age-matched wildtype controls, we devised a predicted miRNA-mRNA interaction network. Among the miRNA-mRNA interactions operating during human neuroblastoma tumorigenesis, we identified miR-204 as a tumor suppressor miRNA that inhibited a subnetwork of oncogenes strongly associated with MYCN-amplified neuroblastoma and poor patient outcome. MYCN bound to the miR-204 promoter and repressed miR-204 transcription. Conversely, miR-204 directly bound MYCN mRNA and repressed MYCN expression. miR-204 overexpression significantly inhibited neuroblastoma cell proliferation in vitro and tumorigenesis in vivo. Together, these findings identify novel tumorigenic miRNA gene networks and miR-204 as a tumor suppressor that regulates MYCN expression in neuroblastoma tumorigenesis. Significance: Network modeling of miRNA-mRNA regulatory interactions in a mouse model of neuroblastoma identifies miR-204 as a tumor suppressor and negative regulator of MYCN.
Carter, DR, Sutton, SK, Pajic, M, Murray, J, Sekyere, EO, Fletcher, J, Beckers, A, De Preter, K, Speleman, F, George, RE, Haber, M, Norris, MD, Cheung, BB & Marshall, GM 2016, 'Glutathione biosynthesis is upregulated at the initiation of MYCN-driven neuroblastoma tumorigenesis', Molecular Oncology, vol. 10, no. 6, pp. 866-878.View/Download from: Publisher's site
© 2016 . The MYCN gene is amplified and overexpressed in a large proportion of high stage neuroblastoma patients and has been identified as a key driver of tumorigenesis. However, the mechanism by which MYCN promotes tumor initiation is poorly understood. Here we conducted metabolic profiling of pre-malignant sympathetic ganglia and tumors derived from the TH-MYCN mouse model of neuroblastoma, compared to non-malignant ganglia from wildtype littermates. We found that metabolites involved in the biosynthesis of glutathione, the most abundant cellular antioxidant, were the most significantly upregulated metabolic pathway at tumor initiation, and progressively increased to meet the demands of tumorigenesis. A corresponding increase in the expression of genes involved in ribosomal biogenesis suggested that MYCN-driven transactivation of the protein biosynthetic machinery generated the necessary substrates to drive glutathione biosynthesis. Pre-malignant sympathetic ganglia from TH-MYCN mice had higher antioxidant capacity and required glutathione upregulation for cell survival, when compared to wildtype ganglia. Moreover, in vivo administration of inhibitors of glutathione biosynthesis significantly delayed tumorigenesis when administered prophylactically and potentiated the anticancer activity of cytotoxic chemotherapy against established tumors. Together these results identify enhanced glutathione biosynthesis as a selective metabolic adaptation required for initiation of MYCN-driven neuroblastoma, and suggest that glutathione-targeted agents may be used as a potential preventative strategy, or as an adjuvant to existing chemotherapies in established disease.
Fabian, J, Opitz, D, Althoff, K, Lodrini, M, Hero, B, Volland, R, Beckers, A, Preter, KD, Decock, A, Patil, N, Abba, M, Kopp-Schneider, A, Astrahantseff, K, Wünschel, J, Pfeil, S, Ercu, M, Künkele, A, Hu, J, Thole, T, Schweizer, L, Mechtersheimer, G, Carter, D, Cheung, BB, Popanda, O, Deimling, AV, Koster, J, Versteeg, R, Schwab, M, Marshall, GM, Speleman, F, Erb, U, Zoeller, M, Allgayer, H, Simon, T, Fischer, M, Kulozik, AE, Eggert, A, Witt, O, Schulte, JH & Deubzer, HE 2016, 'MYCN and HDAC5 transcriptionally repress CD9 to trigger invasion and metastasis in neuroblastoma', Oncotarget, vol. 7, no. 41, pp. 66344-66359.View/Download from: Publisher's site
The systemic and resistant nature of metastatic neuroblastoma renders it largely incurable with current multimodal treatment. Clinical progression stems mainly from the increasing burden of metastatic colonization. Therapeutically inhibiting the migration-invasion-metastasis cascade would be of great benefit, but the mechanisms driving this cycle are as yet poorly understood. In-depth transcriptome analyses and ChIP-qPCR identified the cell surface glycoprotein, CD9, as a major downstream player and direct target of the recently described GRHL1 tumor suppressor. CD9 is known to block or facilitate cancer cell motility and metastasis dependent upon entity. High-level CD9 expression in primary neuroblastomas correlated with patient survival and established markers for favorable disease. Low-level CD9 expression was an independent risk factor for adverse outcome. MYCN and HDAC5 colocalized to the CD9 promoter and repressed transcription. CD9 expression diminished with progressive tumor development in the TH-MYCN transgenic mouse model for neuroblastoma, and CD9 expression in neuroblastic tumors was far below that in ganglia from wildtype mice. Primary neuroblastomas lacking MYCN amplifications displayed differential CD9 promoter methylation in methyl-CpG-binding domain sequencing analyses, and highlevel methylation was associated with advanced stage disease, supporting epigenetic regulation. Inducing CD9 expression in a SH-EP cell model inhibited migration and invasion in Boyden chamber assays. Enforced CD9 expression in neuroblastoma cells transplanted onto chicken chorioallantoic membranes strongly reduced metastasis to embryonic bone marrow. Combined treatment of neuroblastoma cells with HDAC/ DNA methyltransferase inhibitors synergistically induced CD9 expression despite hypoxic, metabolic or cytotoxic stress. Our results show CD9 is a critical and indirectly druggable suppressor of the invasion-metastasis cycle in neuroblastoma.
Sutton, SK, Carter, DR, Kim, P, Tan, O, Arndt, GM, Zhang, XD, Baell, J, Noll, BD, Wang, S, Kumar, N, McArthur, GA, Cheung, BB & Marshall, GM 2016, 'A novel compound which sensitizes BRAF wild-type melanoma cells to vemurafenib in a TRIM16-dependent manner', Oncotarget, vol. 7, no. 32, pp. 52166-52178.View/Download from: Publisher's site
There is an urgent need for better therapeutic options for advanced melanoma patients, particularly those without the BRAFV600E/K mutation. In melanoma cells, loss of TRIM16 expression is a marker of cell migration and metastasis, while the BRAF inhibitor, vemurafenib, induces melanoma cell growth arrest in a TRIM16-dependent manner. Here we identify a novel small molecule compound which sensitized BRAF wild-type melanoma cells to vemurafenib. High throughput, cell-based, chemical library screening identified a compound (C012) which significantly reduced melanoma cell viability, with limited toxicity for normal human fibroblasts. When combined with the BRAFV600E/K inhibitor, vemurafenib, C012 synergistically increased vemurafenib potency in 5 BRAFWT and 4 out of 5 BRAFV600E human melanoma cell lines (Combination Index: CI < 1), and, dramatically reduced colony forming ability. In addition, this drug combination was significantly anti-tumorigenic in vivo in a melanoma xenograft mouse model. The combination of vemurafenib and C012 markedly increased expression of TRIM16 protein, and knockdown of TRIM16 significantly reduced the growth inhibitory effects of the vemurafenib and C012 combination. These findings suggest that the combination of C012 and vemurafenib may have therapeutic potential for the treatment of melanoma, and, that reactivation of TRIM16 may be an effective strategy for patients with this disease.
Beckers, A, Peer, GV, Carter, DR, Mets, E, Althoff, K, Cheung, BB, Schulte, JH, Mestdagh, P, Vandesompele, J, Marshall, GM, De Preter, KD & Speleman, F 2015, 'MYCN-targeting miRNAs are predominantly downregulated during MYCN-driven neuroblastoma tumor formation', Oncotarget, vol. 6, no. 7, pp. 5204-5216.View/Download from: Publisher's site
MYCN is a transcription factor that plays key roles in both normal development and cancer. In neuroblastoma, MYCN acts as a major oncogenic driver through pleiotropic effects controlled by multiple protein encoding genes as well as microRNAs (miRNAs). MYCN activity is tightly regulated at the level of transcription and protein stability through various mechanisms. Like most genes, MYCN is further controlled by miRNAs, but the full complement of all miRNAs implicated in this process has not been determined through an unbiased approach. To elucidate the role of miRNAs in regulation of MYCN, we thus explored the MYCN-miRNA interactome to establish miRNAs controlling MYCN expression levels. We combined results from an unbiased and genome-wide high-throughput miRNA target reporter screen with miRNA and mRNA expression data from patients and a murine neuroblastoma progression model. We identified 29 miRNAs targeting MYCN, of which 12 miRNAs are inversely correlated with MYCN expression or activity in neuroblastoma tumor tissue. The majority of MYCN-targeting miRNAs in neuroblastoma showed a decrease in expression during murine MYCN-driven neuroblastoma tumor development. Therefore, we provide evidence that MYCN-targeting miRNAs are preferentially downregulated in MYCNdriven neuroblastoma, suggesting that MYCN negatively controls the expression of these miRNAs, to safeguard its expression.
Beckers, A, Van Peer, G, Carter, DR, Gartlgruber, M, Herrmann, C, Agarwal, S, Helsmoortel, HH, Althoff, K, Molenaar, JJ, Cheung, BB, Schulte, JH, Benoit, Y, Shohet, JM, Westermann, F, Marshall, GM, Vandesompele, J, De Preter, K & Speleman, F 2015, 'MYCN-driven regulatory mechanisms controlling LIN28B in neuroblastoma', Cancer Letters, vol. 366, no. 1, pp. 123-132.View/Download from: Publisher's site
© 2015 Elsevier Ireland Ltd. LIN28B has been identified as an oncogene in various tumor entities, including neuroblastoma, a childhood cancer that originates from neural crest-derived cells, and is characterized by amplification of the MYCN oncogene. Recently, elevated LIN28B expression levels were shown to contribute to neuroblastoma tumorigenesis via let-7 dependent de-repression of MYCN. However, additional insight in the regulation of LIN28B in neuroblastoma is lacking. Therefore, we have performed a comprehensive analysis of the regulation of LIN28B in neuroblastoma, with a specific focus on the contribution of miRNAs.We show that MYCN regulates LIN28B expression in neuroblastoma tumors via two distinct parallel mechanisms. First, through an unbiased LIN28B-3'UTR reporter screen, we found that miR-26a-5p and miR-26b-5p regulate LIN28B expression. Next, we demonstrated that MYCN indirectly affects the expression of miR-26a-5p, and hence regulates LIN28B, therefore establishing an MYCN-miR-26a-5p-LIN28B regulatory axis. Second, we provide evidence that MYCN regulates LIN28B expression via interaction with the LIN28B promoter, establishing a direct MYCN-LIN28B regulatory axis. We believe that these findings mark LIN28B as an important effector of the MYCN oncogenic phenotype and underline the importance of MYCN-regulated miRNAs in establishing the MYCN-driven oncogenic process.
Carter, DR, Murray, J, Cheung, BB, Gamble, L, Koach, J, Tsang, J, Sutton, S, Kalla, H, Syed, S, Gifford, AJ, Issaeva, N, Biktasova, A, Atmadibrata, B, Sun, Y, Sokolowski, N, Ling, D, Kim, PY, Webber, H, Clark, A, Ruhle, M, Liu, B, Oberthuer, A, Fischer, M, Byrne, J, Saletta, F, Thwe, LM, Purmal, A, Haderski, G, Burkhart, C, Speleman, F, De Preter, K, Beckers, A, Ziegler, DS, Liu, T, Gurova, KV, Gudkov, AV, Norris, MD, Haber, M & Marshall, GM 2015, 'Therapeutic targeting of the MYC signal by inhibition of histone chaperone FACT in neuroblastoma', SCIENCE TRANSLATIONAL MEDICINE, vol. 7, no. 312.View/Download from: Publisher's site
Cheung, BB, Tan, O, Koach, J, Liu, B, Shum, MSY, Carter, DR, Sutton, S, Po'uha, ST, Chesler, L, Haber, M, Norris, MD, Kavallaris, M, Liu, T, O'Neill, GM & Marshall, GM 2015, 'Thymosin-β4 is a determinant of drug sensitivity for Fenretinide and Vorinostat combination therapy in neuroblastoma', Molecular Oncology, vol. 9, no. 7, pp. 1484-1500.View/Download from: Publisher's site
© 2015 Federation of European Biochemical Societies. Retinoids are an important component of neuroblastoma therapy at the stage of minimal residual disease, yet 40-50% of patients treated with 13- cis-retinoic acid (13- cis-RA) still relapse, indicating the need for more effective retinoid therapy. Vorinostat, or Suberoylanilide hydroxamic acid (SAHA), is a potent inhibitor of histone deacetylase (HDAC) classes I & II and has antitumor activity in vitro and in vivo. Fenretinide (4-HPR) is a synthetic retinoid which acts on cancer cells through both nuclear retinoid receptor and non-receptor mechanisms. In this study, we found that the combination of 4-HPR + SAHA exhibited potent cytotoxic effects on neuroblastoma cells, much more effective than 13- cis-RA + SAHA. The 4-HPR + SAHA combination induced caspase-dependent apoptosis through activation of caspase 3, reduced colony formation and cell migration in vitro, and tumorigenicity in vivo. The 4-HPR and SAHA combination significantly increased mRNA expression of thymosin-beta-4 (Tβ4) and decreased mRNA expression of retinoic acid receptor α (RARα). Importantly, the up-regulation of Tβ4 and down-regulation of RARα were both necessary for the 4-HPR + SAHA cytotoxic effect on neuroblastoma cells. Moreover, Tβ4 knockdown in neuroblastoma cells increased cell migration and blocked the effect of 4-HPR + SAHA on cell migration and focal adhesion formation. In primary human neuroblastoma tumor tissues, low expression of Tβ4 was associated with metastatic disease and predicted poor patient prognosis. Our findings demonstrate that Tβ4 is a novel therapeutic target in neuroblastoma, and that 4-HPR + SAHA is a potential therapy for the disease.
Lambertz, I, Kumps, C, Claeys, S, Lindner, S, Beckers, A, Janssens, E, Carter, DR, Cazes, A, Cheung, BB, De Mariano, M, De Bondt, A, De Brouwer, S, Delattre, O, Gibbons, J, Janoueix-Lerosey, I, Laureys, G, Liang, C, Marchall, GM, Porcu, M, Takita, J, Trujillo, DC, Van Den Wyngaert, I, Van Roy, N, Van Goethem, A, Van Maerken, T, Zabrocki, P, Cools, J, Schulte, JH, Vialard, J, Speleman, F & De Preter, K 2015, 'Upregulation of MAPK negative feedback regulators and RET in mutant ALK neuroblastoma: Implications for targeted treatment', Clinical Cancer Research, vol. 21, no. 14, pp. 3327-3339.View/Download from: Publisher's site
© 2015 American Association for Cancer Research. Purpose: Activating ALK mutations are present in almost 10% of primary neuroblastomas and mark patients for treatment with small-molecule ALK inhibitors in clinical trials. However, recent studies have shown that multiple mechanisms drive resistance to these molecular therapies. We anticipated that detailed mapping of the oncogenic ALK-driven signaling in neuroblastoma can aid to identify potential fragile nodes as additional targets for combination therapies. Experimental Design: To achieve this goal, transcriptome profiling was performed in neuroblastoma cell lines with the ALKF1174L or ALKR1275Q hotspot mutations, ALK amplification, or wild-type ALK following pharmacologic inhibition of ALK using four different compounds. Next, we performed cross-species genomic analyses to identify commonly transcriptionally perturbed genes in MYCN/ALKF1174L double transgenic versusMYCN transgenic mouse tumors as compared with the mutant ALKdriven transcriptome in human neuroblastomas. Results: A 77-gene ALK signature was established and successfully validated in primary neuroblastoma samples, in a neuroblastoma cell line with ALKF1174L and ALKR1275Q regulable overexpression constructs and in other ALKomas. In addition to the previously established PI3K/AKT/mTOR, MAPK/ERK, and MYC/MYCN signaling branches, we identified that mutant ALK drives a strong upregulation of MAPK negative feedback regulators and upregulates RET and RET-driven sympathetic neuronal markers of the cholinergic lineage. Conclusions: We provide important novel insights into the transcriptional consequences and the complexity ofmutant ALK signaling in this aggressive pediatric tumor. The negative feedback loop of MAPK pathway inhibitors may affect novel ALK inhibition therapies, whereas mutant ALK induced RET signaling can offer novel opportunities for testing ALK-RET oriented molecular combination therapies.
Sun, Y, Bell, JL, Carter, D, Gherardi, S, Poulos, RC, Milazzo, G, Wong, JWH, Al-Awar, R, Tee, AE, Liu, PY, Liu, B, Atmadibrata, B, Wong, M, Trahair, T, Zhao, Q, Shohet, JM, Haupt, Y, Schulte, JH, Brown, PJ, Arrowsmith, CH, Vedadi, M, MacKenzie, KL, Hüttelmaier, S, Perini, G, Marshall, GM, Braithwaite, A & Liu, T 2015, 'WDR5 supports an N-myc transcriptional complex that drives a protumorigenic gene expression signature in neuroblastoma', Cancer Research, vol. 75, no. 23, pp. 5143-5154.View/Download from: Publisher's site
© 2015 American Association for Cancer Research. MYCN gene amplification in neuroblastoma drives a gene expression program that correlates strongly with aggressive disease. Mechanistically, trimethylation of histone H3 lysine 4 (H3K4) at target gene promoters is a strict prerequisite for this transcriptional program to be enacted. WDR5 is a histone H3K4 presenter that has been found to have an essential role in H3K4 trimethylation. For this reason, in this study, we investigated the relationship between WDR5-mediated H3K4 trimethylation and N-Myc transcriptional programs in neuroblastoma cells. N-Myc upregulated WDR5 expression in neuroblastoma cells. Gene expression analysis revealed that WDR5 target genes included those with MYC-binding elements at promoters such as MDM2. We showed that WDR5 could form a protein complex at the MDM2 promoter with N-Myc, but not p53, leading to histone H3K4 trimethylation and activation of MDM2 transcription. RNAi-mediated attenuation of WDR5 upregulated expression of wild-type but not mutant p53, an effect associated with growth inhibition and apoptosis. Similarly, a small-molecule antagonist of WDR5 reduced N-Myc/WDR5 complex formation, N-Myc target gene expression, and cell growth in neuroblastoma cells. In MYCN-transgenic mice, WDR5 was overexpressed in precancerous ganglion and neuroblastoma cells compared with normal ganglion cells. Clinically, elevated levels of WDR5 in neuroblastoma specimens were an independent predictor of poor overall survival. Overall, our results identify WDR5 as a key cofactor for N-Myc-regulated transcriptional activation and tumorigenesis and as a novel therapeutic target for MYCN-amplified neuroblastomas.
Kim, PY, Tan, O, Diakiw, SM, Carter, D, Sekerye, EO, Wasinger, VC, Liu, T, Kavallaris, M, Norris, MD, Haber, M, Chesler, L, Dolnikov, A, Trahair, TN, Cheung, NK, Marshall, GM & Cheung, BB 2014, 'Identification of plasma Complement C3 as a potential biomarker for neuroblastoma using a quantitative proteomic approach', Journal of Proteomics, vol. 96, pp. 1-12.View/Download from: Publisher's site
The majority of patients diagnosed with neuroblastoma present with aggressive disease. Improved detection of neuroblastoma cancer cells following initial therapy may help in stratifying patient outcome and monitoring for relapse. To identify potential plasma biomarkers, we utilised a liquid chromatography-tandem mass spectrometry-based proteomics approach to detect differentially-expressed proteins in serum from TH-MYCN mice. TH-MYCN mice carry multiple copies of the human MYCN oncogene in the germline and homozygous mice for the transgene develop neuroblastoma in a manner resembling the human disease. The abundance of plasma proteins was measured over the course of disease initiation and progression. A list of 86 candidate plasma biomarkers was generated. Pathway analysis identified significant association of these proteins with genes involved in the complement system. One candidate, complement C3 protein, was significantly enriched in the plasma of TH-MYCN+/+ mice at both 4 and 6weeks of age, and was found to be elevated in a cohort of human neuroblastoma plasma samples, compared to healthy subjects. In conclusion, we have demonstrated the suitability of the TH-MYCN+/+ mouse model of neuroblastoma for identification of novel disease biomarkers in humans, and have identified Complement C3 as a candidate plasma biomarker for measuring disease state in neuroblastoma patients. Biological significance: This study has utilised a unique murine model which develops neuroblastoma tumours that are biologically indistinguishable from human neuroblastoma. This animal model has effectively allowed the identification of plasma proteins which may serve as potential biomarkers of neuroblastoma. Furthermore, the label-free ion count quantitation technique which was used displays significant benefits as it is less labour intensive, feasible and accurate. We have been able to successfully validate this approach by confirming the differential abundance of two different plasma prot...
Marshall, GM, Carter, DR, Cheung, BB, Liu, T, Mateos, MK, Meyerowitz, JG & Weiss, WA 2014, 'The prenatal origins of cancer', Nature Reviews Cancer, vol. 14, no. 4, pp. 277-289.View/Download from: Publisher's site
The concept that some childhood malignancies arise from postnatally persistent embryonal cells has a long history. Recent research has strengthened the links between driver mutations and embryonal and early postnatal development. This evidence, coupled with much greater detail on the cell of origin and the initial steps in embryonal cancer initiation, has identified important therapeutic targets and provided renewed interest in strategies for the early detection and prevention of childhood cancer. © 2014 Macmillan Publishers Limited. All rights reserved.
Sutton, SK, Koach, J, Tan, O, Liu, B, Carter, DR, Wilmott, JS, Yosufi, B, Haydu, LE, Mann, GJ, Thompson, JF, Long, GV, Liu, T, McArthur, G, Zhang, XD, Scolyer, RA, Cheung, BB & Marshall, GM 2014, 'TRIM16 inhibits proliferation and migration through regulation of interferon beta 1 in melanoma cells', Oncotarget, vol. 5, no. 20, pp. 10127-10139.View/Download from: Publisher's site
High basal or induced expression of the tripartite motif protein, TRIM16, leads to reduce cell growth and migration of neuroblastoma and skin squamous cell carcinoma cells. However, the role of TRIM16 in melanoma is currently unknown. TRIM16 protein levels were markedly reduced in human melanoma cell lines, compared with normal human epidermal melanocytes due to both DNA methylation and reduced protein stability. TRIM16 knockdown strongly increased cell migration in normal human epidermal melanocytes, while TRIM16 overexpression reduced cell migration and proliferation of melanoma cells in an interferon beta 1 (IFNβ1)-dependent manner. Chromatin immunoprecipitation assays revealed TRIM16 directly bound the IFNβ1 gene promoter. Low level TRIM16 expression in 91 melanoma patient samples, strongly correlated with lymph node metastasis, and, predicted poor patient prognosis in a separate cohort of 170 melanoma patients with lymph node metastasis. The BRAF inhibitor, vemurafenib, increased TRIM16 protein levels in melanoma cells in vitro, and induced growth arrest in BRAF-mutant melanoma cells in a TRIM16-dependent manner. High levels of TRIM16 in melanoma tissues from patients treated with Vemurafenib correlated with clinical response. Our data, for the first time, demonstrates TRIM16 is a marker of cell migration and metastasis, and a novel treatment target in melanoma.