Neuroblastoma Survival Research Articles

Renal cell carcinoma after neuroblastoma: A case study and review of the literature.

We present a case of renal cell carcinoma (RCC) arising in a 26-year-old patient with a history of neuroblastoma. RCC after a previous diagnosis of neuroblastoma is very uncommon, and there have only been 23 reported cases. Using the results of our patient workup, we hoped to determine whether there was a genetic predisposition or iatrogenic cause for the RCC. There is no clear explanation why neuroblastoma survivors are prone to developing RCC. However, genetic predisposition and previous treatment likely play a role. Since there have been few cases described, and few investigations into the genetics of this subtype of RCC, it remains important for individual cases to be added to the literature of this recently described and rare entity.

Health-related quality of life in young survivors of childhood cancer.

Childhood cancer and its treatment may affect health-related quality of life (HRQoL) in childhood cancer survivors, but population-based studies in young survivors are scarce. We aimed to: (1) compare HRQoL between young survivors and population norms and (2) find factors that influence parent-reported HRQoL in survivors. As part of the Swiss Childhood Cancer Survivor Study, a questionnaire was mailed to parents of survivors aged 8-16 years, registered in the Swiss Childhood Cancer Registry, ≥5 years after diagnosis. We used the KIDSCREEN-27 instrument to compare self- and parent-reported HRQoL between survivors (N = 425) and standardized norms in the five dimensions of physical well-being, psychological well-being, autonomy, peers and school environment (mean = 50, SD = 10). We then used multivariable linear regressions to test the influence of socio-demographic and cancer-related factors on HRQoL. Self-reported physical well-being was comparable to norms. Other HRQoL dimensions were higher than norms, with the highest mean = 52.2 (p < 0.001) for school environment. Parent-reported HRQoL in survivors was comparable to population norms; only physical well-being was lower (mean = 47.1, p < 0.001), and school environment was higher (mean = 51.1, p = 0.035). Parent-reported HRQoL was lower for survivors of CNS tumors (physical well-being: β = -5.27, p = 0.007; psychological well-being: β = -4.39, p = 0.044; peers β = -5.17, p = 0.028), survivors of neuroblastoma (psychological well-being β = -5.20, p = 0.047), and survivors who had had a relapse (physical well-being β = -5.41, p = 0.005). Assessing HRQoL during follow-up care, with a focus on physical well-being, specific diagnoses (e.g., CNS tumor) and late complications (e.g., relapse) might help to early identify problems and offer support to survivors with reduced HRQoL.

Alternative NHEJ Pathway Components Are Therapeutic Targets in High-Risk Neuroblastoma

In neuroblastoma, MYCN genomic amplification and segmental chromosomal alterations including 1p or 11q loss of heterozygocity and/or 17q gain are associated with progression and poor clinical outcome. Segmental alterations are the strongest predictor of relapse and result from unbalanced translocations attributable to erroneous repair of chromosomal breaks. Although sequence analysis of affected genomic regions suggests that these errors arise by nonhomologous end-joining (NHEJ) of DNA double-strand breaks (DSB), abnormalities in NHEJ have not been implicated in neuroblastoma pathogenesis. On this basis, the hypothesis that an error-prone mechanism of NHEJ is critical for neuroblastoma cell survival was tested. Plasmid-based DSB repair assays demonstrated efficient NHEJ activity in human neuroblastoma cells with repair products that were error-prone relative to nontransformed cells. Neuroblastoma cells derived from tumorigenic neuroblastic phenotypes had differential DNA repair protein expression patterns compared with nontumorigenic cells. Tumorigenic neuroblastoma cells were deficient in DNA ligase IV (Lig4) and Artemis (DCLRE1C), mediators of canonical NHEJ. Conversely, enzymes required for an error-prone alternative NHEJ pathway (alt-NHEJ), DNA Ligase IIIα (Lig3), DNA Ligase I (Lig1), and PARP1 protein were upregulated. Inhibition of Lig3 and Lig1 led to DSB accumulation and cell death, linking alt-NHEJ to cell survival in neuroblastoma. Neuroblastoma cells demonstrated sensitivity to PARP1 inhibition (PARPi) that paralleled PARP1 expression. In a dataset of human neuroblastoma patient tumors, overexpression of genes encoding alt-NHEJ proteins associated with poor survival. These findings provide an insight into DNA repair fidelity in neuroblastoma and identify components of the alt-NHEJ pathway as promising therapeutic targets.

Two independent epigenetic biomarkers predict survival in neuroblastoma.

BackgroundNeuroblastoma (NB) is the most common extracranial pediatric solid tumor with a highly variable clinical course, ranging from spontaneous regression to life-threatening disease. Survival rates for high-risk NB patients remain disappointingly low despite multimodal treatment. Thus, there is an urgent clinical need for additional biomarkers to improve risk stratification, treatment management, and survival rates in children with aggressive NB.ResultsUsing gene promoter methylation analysis in 48 neuroblastoma tumors with microarray technology, we found a strong association between survival and gene promoter hypermethylation (P = 0.036). Hypermethylation of 70 genes significantly differentiated high-risk survivor patients from those who died during follow-up time. Sixteen genes with relevant roles in cancer biology were further validated in an additional cohort of 83 neuroblastoma tumors by bisulfite pyrosequencing. High promoter methylation rates of these genes were found in patients with metastatic tumors (either stage metastatic (M) or metastatic special (MS)), 18 months or older at first diagnosis, MYCN amplification, relapsed, and dead. Notably, the degree of methylation of retinoblastoma 1 (RB1) and teratocarcinoma-derived growth factor 1 (TDGF1) predicts event-free and overall survival independently of the established risk factors. In addition, low RB1 mRNA expression levels associate with poor prognosis suggesting that promoter methylation could contribute to the transcriptional silencing of this gene in NB.ConclusionsWe found a new epigenetic signature predictive for NB patients’ outcome: the methylation status of RB1 and TDGF1 associate with poorer survival. This information is useful to assess prognosis and improve treatment selection.Electronic supplementary materialThe online version of this article (doi:10.1186/s13148-015-0054-8) contains supplementary material, which is available to authorized users.

Long-term efficacy of current thyroid prophylaxis and future perspectives on thyroid protection during 131I-metaiodobenzylguanidine treatment in children with neuroblastoma.

Treatment with (131)I-MIBG is associated with significant thyroid damage. This study was undertaken to investigate the long-term efficacy of current thyroid prophylaxis, to explore the relationship between thyroid dysfunction and thyroid volume after exposure to (131)I-MIBG and to evaluate the possible negative effects of (131)I(-) on the parathyroid glands. Of 81 long-term surviving patients with neuroblastoma treated with (131)I-MIBG during the period 1999-2012, 24 were finally evaluated. Patients received thyroxine (T4), methimazole and potassium iodide as thyroid protection. In all patients (para)thyroid function was evaluated and ultrasound investigation of the (para)thyroid gland(s) was performed. Thyroid dysfunction was defined as a plasma thyrotropin concentration >5.0mU/L (thyrotropin elevation, TE) or as the use of T4 at the time of follow-up. Hyperparathyroidism was defined as a serum calcium concentration above the age-related reference range in combination with an inappropriately high parathyroid hormone level. At a median follow-up of 9.0years after (131)I-MIBG treatment, thyroid disorders were seen in 12 patients (50%; 9 with TE, 5 with a thyroid nodule and 1 patient was subsequently diagnosed with differentiated thyroid carcinoma). No significant risk factors for the occurrence of thyroid damage could be identified. In 14 of 21 patients (67%) in whom thyroid volume could be determined, the volume was considered small (<-2SD) for age and gender. Patients treated with T4 at the time of follow-up had significantly smaller thyroid volumes for age than patients without T4 treatment (p = 0.014). None of the patients was diagnosed with hyperparathyroidism. Thyroid protection during treatment with (131)I-MIBG needs attention and must be further improved, as thyroid disorders are still frequently seen despite current thyroid prophylaxis. Reduced thyroid volume in neuroblastoma survivors may be related to previous (131)I-MIBG therapy or current T4 treatment. No deleterious effects of (131)I-MIBG on the parathyroid glands could be found.

Aurora kinase A is superior to Ki67 as a prognostic indicator of survival in neuroblastoma.

To compare the expression and prognostic value of the cell cycle markers Aurora kinase A (AURKA) and Ki67 in neuroblastoma, because AURKA expression levels have greater prognostic significance than those of Ki67 in some cancers. Eighty-eight neuroblastomas were immunostained with anti-AURKA and Ki67 antibodies. Digitally scanned slides were scored using imaging analysis software. Median AURKA and Ki67 proliferation indices (PIs) were 1.5% and 26%, respectively. Higher than median AURKA and Ki67 levels were detected in the neuroblastomas from patients belonging to the high-risk group, those with MYCN amplification, and those with unfavourable pathology, including a high mitosis-karyorrhexis index (MKI). High AURKA and Ki67 levels were significantly associated with shorter overall survival (OS) and event-free survival (EFS) in univariate analyses. In multivariate analyses, high AURKA level was associated with significantly shorter OS and EFS, independently of risk group, and of MYCN amplification and MKI. High Ki67 level was not associated with shorter OS or EFS after adjustment for risk group or MYCN amplification and MKI. High AURKA and Ki67 levels were associated with adverse prognostic factors and shorter survival, but AURKA provides more valuable prognostic information than Ki67 in neuroblastoma.

Highlights from Recent Cancer Literature

Highlights from Recent Cancer Literature

Immune-based therapies for childhood cancer.

After decades of research, immunotherapies for cancer are demonstrating increasing success. These agents can amplify existent antitumour immunity or induce durable antitumour immune responses in a wide array of cancers. The spectrum of immunotherapeutics is broad, spanning monoclonal antibodies and their derivatives, tumour vaccines, and adoptive therapies using T cells and natural killer cells. Only a small number of immunotherapies have been tested in paediatric cancers, but impressive antitumour effects have already been observed. Mononclonal antibodies targeting GD2 that induce antibody-dependent cell-mediated cytotoxicity improve survival in high-risk neuroblastoma. Bi-specific monoclonal antibodies that simultaneously target CD19 and activate T cells can induce remission in acute B-cell lymphoblastic leukaemia (B-ALL) and adoptive immunotherapy using T cells genetically engineered to express chimeric antigen receptors targeting CD19 induce impressive responses in B-ALL. Efforts are underway to generate and test new immunotherapies in a wider array of paediatric cancers. Major challenges include a need to identify immunotherapy targets on the most lethal childhood cancers, to expand availability of technology-intense platforms, such as adoptive cell therapy, to optimize management of novel toxicities associated with this new class of cancer therapies and to determine how best to incorporate these therapies into standard treatment paradigms.

Childhood cancer incidence and survival 1985-2009, Khon Kaen, Thailand.

The Khon Kaen Cancer Registry (KKCR) was established in 1984. Previous population-based incidences and survivals of childhood cancer in Thailand were determined using a short cancer registration period. Data were retrieved of all children residing in Khon Kaen, between 0-15 years, diagnosed as having cancer and registered in the KKCR (1985-2009). The follow-up censored date was December 31, 2012. The childhood cancers were classified into 12 diagnostic groups, according to the International Classification of Childhood Cancer. The incidence was calculated by the standard method. Survival of childhood cancer was investigated using the KKCR population-based registration data and overall survival calculated using the Kaplan Meier method. In the study period, 912 newly diagnosed cases of childhood cancer were registered. The respective mean and median age was 6.4 (SD=4.6) and 6 (0-14) years. The age-peak for incidence was 0-4 years. The age-standardized rate (ASR) was 83 per million. Leukemia was the most common cancer (N=360, ASR 33.8) followed by neoplasms of the central nervous system (CNS, N=150, ASR 12.8) and lymphoma (N=79, ASR 7.0). The follow-up duration totaled 101,250 months. The death rate was 1.11 per 100 person-months (95%CI: 1.02 -1.20). The 5-year overall survival was 52% (95%CI: 53-56.9) for all cancers. The respective 5-year overall survival for (1) acute lymphoblastic leukemia (ALL), (2) acute non-lymphoblastic leukemia (ANLL), (3) lymphoma, (4) germ cell tumors, (5) renal tumors, (6) retinoblastoma, (7) soft tissue tumors, (8) CNS tumors, (9) bone tumors, (10) liver tumors, and (11) neuroblastoma was (1) 51%, (2) 37%, (3) 63%, (4) 74%, (5) 67%, (6) 55%, (7) 46%, (8) 44%, (9) 36%, (10) 34%, and (11) 25%. The incidence of childhood cancer is lower than those of western countries. Respective overall survival for ALL, lymphoma, renal tumors, liver tumors, retinoblastoma, soft tissue tumors is lower than that reported in developed countries while survival for CNS tumors, neuroblastoma and germ cell tumors is comparable.

Abstract B39: IGF-IR regulation of osteoclastogenesis in neuroblastoma bone metastasis

Abstract Neuroblastoma is a tumor that is responsible for 15% of cancer deaths in children and has a mortality rate greater than 90% percent upon metastasis to bone. Due to the high mortality rate and lack of effective treatment options, our laboratory is investigating potential new therapeutic targets in neuroblastoma bone metastases. Since, insulin-like growth factor I (IGF-I) production in the bone promotes neuroblastoma growth and survival, our hypothesis is that inhibiting IGF-I receptor (IGF-IR) will prevent metastasis of this tumor to bone. Furthermore, neuroblastoma with increased IGF-IR expression induces osteoclasts to differentiate, with the percentage of differentiated osteoclasts in vitro increasing to 30% in the presence of conditioned medium from high IGF-IR expressing cells. The addition of a monoclonal, neutralizing antibody to the IGF-IR decreases the percentage of mature osteoclasts formed by over 50%. We are testing the hypothesis that IGF-IR activation in neuroblastoma alters the levels of RANK, RANKL and OPG, proteins involved in osteoclast differentiation. This alteration in RANK, RANKL, and OPG may cause osteoclast differentiation and osteolysis. Neuroblastoma cells with high IGF-IR protein levels express low levels of the decoy receptor OPG, while some lines also express high levels of RANKL. Therefore, we are currently testing the effect of IGF-IR inhibition on RANKL and OPG levels and subsequent effect on osteolysis. Citation Format: Chris A. Rivera-Pintado, Yufei Shan, Cynthia M. van Golen. IGF-IR regulation of osteoclastogenesis in neuroblastoma bone metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Pediatric Cancer at the Crossroads: Translating Discovery into Improved Outcomes; Nov 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;74(20 Suppl):Abstract nr B39.

Abstract 3607: Modulation of neuroblastoma metabolism by an extracellular chaperone SPARC

Abstract Secreted Protein Acidic and Rich in Cysteine (SPARC) is an extracellular matrix protein with diverse biologic functions. We have shown that SPARC is a scavenger chaperone that regulates matrix degradation, remodeling, and repair. We have also demonstrated that SPARC blocks angiogenesis and inhibits the growth of several cancers, including neuroblastoma. In this study we examined how extracellular chaperone SPARC affects neuroblastoma tumor progression. The chaperone properties of SPARC were characterized with cell biology methods and Surface Plasmon Resonance. Neuroblastoma cell lines derived from the high-risk tumors were treated with SPARC. The effect of SPARC on cancer cell metabolism was characterized using the Seahorse analyzer with and without inhibitors of glycolysis and mitochondrial oxidation. Standard cell culture techniques were used to assess the effect of SPARC-induced metabolic changes on neuroblastoma cell proliferation and survival. We demonstrate that SPARC functions as a chaperone, shuttling extracellular ligands between cellular and extracellular tissue compartments. We characterize physical and biological properties of SPARC-mediated ligand transport, and further demonstrate that as a chaperone, SPARC can significantly affect tissue physiology, to the degree of altering cell metabolism. We show that in addition to inhibition of neuroblastoma progression in vivo, SPARC-induced changes in cellular metabolism severely impair neuroblastoma growth and survival in vitro. This novel mechanism of metabolism regulation by an extracellular chaperone SPARC may lead to additional insights into the therapy of neuroblastoma. Citation Format: Alexandre Chlenski, Marija Dobratic, Helen R. Salwen, Mark Applebaum, Susan L. Cohn. Modulation of neuroblastoma metabolism by an extracellular chaperone SPARC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3607. doi:10.1158/1538-7445.AM2014-3607

Abstract 4614: Identification of two salinomycin binding targets in neuroblastoma

Abstract Salinomycin, a widely used anti-coccidial agent isolated from a Streptomyces albus strain, was recently identified from a library of 16,000 natural and commercial chemical compounds based on its highly selective inhibitory effect on breast cancer stem cells (CSCs), with more than 100-fold greater potency than paclitaxel. Salinomycin also exhibits cytotoxic effects on other types of cancer cells and CSCs and overcomes drug resistance. However, the exact mechanism of salinomycin, especially its direct binding target(s), and its effects on neuroblastoma (NB) are yet not known. NB is one of the most common solid tumors in children and infants, and a leading cause of mortality in children. Despite advances in diagnosis and treatment, currently, 35% of patients with NB remain incurable. In addition, the majority survivors of NB suffer from long-term side effects of current therapies and are at risk for disease relapse or getting a second, different cancer. More effective therapies are pressingly needed. Since the existence of CSCs in human NB cell lines and NB tumors from patients has been well documented, and has been closely associated with chemoresistance or tumor relapse, therapeutic targeting of NB CSCs may be a critical novel approach to conquer the challenges in NB therapy. Aiming to improve NB therapy, we examined the efficacy and mechanism of salinomycin in human NB cells. Our study showed that salinomycin markedly inhibits NB cell proliferation and tumorsphere formation. Treatment of salinomycin induced G2 cell cycle arrest with an up-regulation of Cyclin A and a down-regulation of p21 protein levels. We further identified TIF1β and NCL as novel direct binding proteins of salinomycin by using comprehensive methods, including chemical proteomics and functional genomics. Furthermore, by analyzing tumor samples from a cohort of 88 NB patients, we found that the elevated level of these proteins is associated with poor prognosis for NB patients. Thus, our results indicate salinomycin is a potential new therapeutic agent and TIF1β and NCL as new therapeutic targets/biomakers for NB therapy. Citation Format: Shuang Zhou, Fengfei Wang, Shi-Hua Xiang, Eric T. Wong, Wallace W. Muhonen, Ekokobe Fonkem, Tze-chen Hsieh, David W. Li, Ruiwen Zhang, John B. Shabb, Joseph M. Wu, Min Wu, Erxi Wu. Identification of two salinomycin binding targets in neuroblastoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4614. doi:10.1158/1538-7445.AM2014-4614

C10ORF10/DEPP, a transcriptional target of FOXO3, regulates ROS-sensitivity in human neuroblastoma.

BackgroundFOXO transcription factors control cellular levels of reactive oxygen species (ROS) which critically contribute to cell survival and cell death in neuroblastoma. In the present study we investigated the regulation of C10orf10/DEPP by the transcription factor FOXO3. As a physiological function of C10orf10/DEPP has not been described so far we analyzed its effects on cellular ROS detoxification and death sensitization in human neuroblastoma cells.MethodsThe effect of DEPP on cellular ROS was measured by catalase activity assay and live cell fluorescence microscopy using the ROS-sensitive dye reduced MitoTracker Red CM-H2XROS. The cellular localization of DEPP was determined by confocal microscopy of EYFP-tagged DEPP, fluorescent peroxisomal- and mitochondrial probes and co-immunoprecipitation of the PEX7 receptor.ResultsWe report for the first time that DEPP regulates ROS detoxification and localizes to peroxisomes and mitochondria in neuroblastoma cells. FOXO3-mediated apoptosis involves a biphasic ROS accumulation. Knockdown of DEPP prevented the primary and secondary ROS wave during FOXO3 activation and attenuated FOXO3- and H2O2-induced apoptosis. Conditional overexpression of DEPP elevates cellular ROS levels and sensitizes to H2O2 and etoposide-induced cell death. In neuronal cells, cellular ROS are mainly detoxified in peroxisomes by the enzyme CAT/catalase. As DEPP contains a peroxisomal-targeting-signal-type-2 (PTS2) sequence at its N-terminus that allows binding to the PEX7 receptor and import into peroxisomes, we analyzed the effect of DEPP on cellular detoxification by measuring enzyme activity of catalase. Catalase activity was reduced in DEPP-overexpressing cells and significantly increased in DEPP-knockdown cells. DEPP directly interacts with the PEX7 receptor and localizes to the peroxisomal compartment. In parallel, the expression of the transcription factor peroxisome proliferator-activated receptor gamma (PPARG), a critical regulator of catalase enzyme activity, was strongly upregulated in DEPP-knockdown cells.ConclusionThe combined data indicate that in neuroblastoma DEPP localizes to peroxisomes and mitochondria and impairs cellular ROS detoxification, which sensitizes tumor cells to ROS-induced cell death.Electronic supplementary materialThe online version of this article (doi:10.1186/1476-4598-13-224) contains supplementary material, which is available to authorized users.

Ototoxicity: a worrying problem for survivors of high-risk neuroblastoma

Landier et al. report the results of a COG study about ototoxicity in children with neuroblastoma. The authors present the results of audiologic testing graded by four different scales [American Speech-Language-Hearing Association; Brock; Chang; and Common Terminology Criteria for Adverse Events version 3 (CTCAEv3)], in 333/489 evaluable neuroblastoma patients, treated with cisplatin alone (400 mg/m 2 ) or after cisplatin (400 mg/m 2 ) plus carboplatin (1,700 mg/m 2 ) according to COG A3973 protocol. (Their conclusions are that severe hearing loss is prevalent among children with highrisk neuroblastoma. Exposure to cisplatin combined with myeloablative carboplatin significantly increases risk. The Brock scale underestimates severe hearing loss and should be used with caution in this setting). Hearing loss from cisplatin or carboplatin is permanent and may have a delayed onset, with progressive loss occurring many years after completion of therapy. Young children are particularly sensitive to this adverse effect. Children treated for high-risk neuroblastoma, for example, are likely to sustain moderate to severe therapy-related hearing loss, with high potential for difficulties in speech discrimination and language acquisition, diminished academic achievement compared with peers with normal hearing, the potential for lifelong impairment of language and academic skills, and diminished quality of life. Although often underappreciated, even hearing loss restricted to high frequency ranges (4,000-8,000 Hz) have a significant impact on language development, verbal abilities, and reasoning skills in young children. This is of particular concern because the ototoxic effects are concurrent with the developmental period in which the process of acquiring speech and language skills is so critical. The reported incidence of cisplatin ototoxicity in children ranges from 26% to over 90% with the variation influenced by treatment and patient-related factors, but it also depends to some extent on the grading system used. The absence of a practical and uniform system for grading ototoxicity has impeded assessment of results in prior studies, and another problem is that full compliance with complete audiological testing in these young patients remains a challenge.

Phase I/IIa multicenter trial for high-risk and recurrent neuroblastoma: Anti-GD2 antibody (ch14.18) immunotherapies using M-CSF or G-CSF.

TPS10095 Background: Anti-GD2 antibody, ch14.18, with GM-CSF or IL2, has significantly improved disease free survival in high-risk neuroblastoma. ch14.18 generally induces cell lysis through the pr...

Identification and clinical prognosis of salinomycin binding targets in neuroblastoma (LB613)

Salinomycin, a widely used anti‐coccidial agent, was recently identified from a library of 16,000 natural and commercial chemical compounds based on its highly selective inhibitory effect on breast cancer stem cells (CSCs), with more than 100‐fold greater potency than paclitaxel. Salinomycin also exhibits cytotoxic effects on other types of cancer cells and CSCs and overcomes drug resistance. However, the exact mechanism of salinomycin, especially its direct binding target(s), and its effects on Neuroblastoma (NB) are yet not known.NB is a common solid tumor and a leading cause of mortality in children. Currently, 35% of patients with NB remain incurable. In addition, the majority survivors of NB suffer from long‐term side effects of current therapies and are at risk for disease relapse or getting a second, different cancer. More effective therapies are pressingly needed. Since the existence of CSCs in human NB cell lines and NB tumors has been well documented, and has been closely associated with chemoresistance or tumor relapse, therapeutic targeting of NB CSCs may be a critical novel approach for NB therapy.Aiming to improve NB therapy, we examined the efficacy and mechanism of salinomycin in human NB cells. Our study showed that salinomycin markedly inhibits NB cell proliferation and tumorsphere formation. Treatment of salinomycin induced G2 cell cycle arrest with an up‐regulation of Cyclin A and a down‐regulation of p21 protein levels. We further identified TIF1β and NCL as novel direct binding targets of salinomycin by using comprehensive methods, including chemical proteomics and functional genomics. Furthermore, by analyzing tumor samples from a cohort of 88 NB patients, we found that the elevated level of either TIF1β or NCL is associated with poor prognosis for NB patients. Thus, our results indicate salinomycin is a potential new therapeutic agent and TIF1β and NCL as new therapeutic targets/biomarkers for NB therapy.Grant Funding Source: Supported by ND EPSCoR, NDSU, NIH P20 RR020151, NIGMS (P20 GM103505 and P30 GM103332‐01)

Expression of the monocyte chemotactic protein-1-induced protein 1 decreases human neuroblastoma cell survival

The importance of monocyte chemotactic protein-1-induced protein 1 (MCPIP1) in the negative regulation of inflammatory reactions has already been extensively studied. However, its role in cancer is not yet established. We studied MCPIP1 gene expression in primary human neuroblastomas and several neuroblastoma cell lines. Our results showed a lack of MCPIP1 expression in primary neuroblastoma tumors. Moreover, it was found that the low expression of the protein measured in human neuroblastoma cell lines might be important for neuroblastoma survival, since enforced MCPIP1 gene expression in human neuroblastoma BE(2)-C cells caused a significant decrease in neuroblastoma cell viability and proliferation.

Functional Retroperitoneal Neuroblastoma and Ganglioneuroblastoma in Adults

ABSTRACTGanglioneuroblastoma and neuroblastoma are malignant catecholamine-secreting tumors arising from neural crest tissue. Thirty-eight cases of functional retroperitoneal extra-adrenal neuroblastoma and ganglioneuroblastoma in adults are reported in English literature. These tumors behave more aggressively in adults compared to children and have very poor prognosis. We report four adults who presented with retroperitoneal extraadrenal ganglioneuroblastoma and neuroblastoma between 2008 and 2012. Median age at presentation was 28.5 years (21-40 years). One patient was in late stages of pregnancy. Three patients presented with local pressure effects from the tumor and one patients manifested clinical features of catecholamine excess. Biochemical analysis showed raised urinary dopamine excretion in all our patients. Urinary noradrenaline and metanephrines were elevated in three cases. Computed tomography/magnetic resonance imaging/meta-iodobenzylguanidine (CT/MRI/MIBG) confirmed stage IIB disease in one patient, stage III disease one patient and stage IV in two cases. Preoperative biopsy confirmed diagnosis in two cases. Complete surgical resection was performed in patient with stage II B disease and partial surgical resection was feasible in other cases. Histological diagnosis was ganglioneuroblastoma in two cases and neuroblastoma in two cases. Meta-iodobenzylglunidine/radiotherapy and chemotherapy were used for residual disease. Urinary catecholamines and metanephrines normalized after complete tumor resection but remained elevated in residual disease. Survival for neuroblastoma was 15 and 21 months after the surgery and 29 months for ganglioneuroblastoma. One patient remains free of disease at 6 months. Functioning ganglioneuroblastoma and neuroblastoma are rare in adults and only one other case ganglioneuroblastoma in pregnancy is reported in the world literature. Measurement of plasma/urinary excretion of catecholamines and their precursors and metabolites is helpful for diagnosis of neuroblastoma and ganglioneuroblastoma. Due to late presentation surgical resection is unlikely to be curative. Three of our patients developed local recurrence of disease. Urinary/plasma assay are used for monitoring disease relapse. Long-term survival is unusual.

PGK1 as Predictor of CXCR4 Expression, Bone Marrow Metastases and Survival in Neuroblastoma

Background and AimA close relationship between phosphoglycerate kinase 1 (PGK1) and the CXCR4/SDF1 axis (chemokine receptor 4/stromal cell derived factor 1) has been shown for several cancers. However, the role of PGK1 has not been investigated for neuroblastoma, and PGK1 might be a therapeutic target for this tumor entity. The aim of the current study was to evaluate the role of PGK1 expression in neuroblastoma patients, to determine the impact of PGK1 expression levels on survival, and to correlate PGK1 expression with CXCR4 expression and bone marrow dissemination.Materials and MethodsSamples from 22 patients with neuroblastoma that were surgically treated at the University Medical Center Hamburg-Eppendorf were evaluated for expression of PGK1 and CXCR4 using immunohistochemistry. Results were correlated with clinical parameters, metastases and outcome of patients. Immunocytochemistry, proliferation and expression analysis of CXCR4 and PGK1 were performed in neuroblastoma cell lines.ResultsPGK1 is expressed in neuroblastoma cells. PGK1 expression is significantly positively correlated with CXCR4 expression and tumor dissemination to the bone marrow. Moreover the expression of PGK1 is significantly associated with a negative impact on survival in patients with neuroblastoma. PGK1 is downregulated by inhibition of CXCR4 in neuroblastoma cells.ConclusionPGK1 appears to play an important role for neuroblastoma, predicting survival and tumor dissemination. Further in vivo studies outstanding, it is a candidate target for novel therapeutic strategies.

Radiation Therapy to the Primary and Postinduction Chemotherapy MIBG-Avid Sites in High-Risk Neuroblastoma

Radiation Therapy to the Primary and Postinduction Chemotherapy MIBG-Avid Sites in High-Risk Neuroblastoma