Spontaneous Regression Of Neuroblastoma Research Articles

Identification of genes associated with spontaneous regression of neuroblastoma

AbstractThe study of target genes for the spontaneous regression phenomenon of neuroblastoma (NB) is still unclear. Common differentially expressed genes (DEGs) were identified by differential expression analysis in both public databases for the stage 4 death group and stage 4S survival group. The DEGs were ranked by constructing protein–protein interaction (PPI) network as well as calculating betweenness centrality (BC) values, and the relationship with NB prognosis was determined by performing univariate analysis, multifactor analysis, and lasso regression analysis on the top 10 genes and possibly correlated with spontaneous regression of NB. We identified a total of 173 DEGs, including 143 upregulated genes and 30 downregulated genes. PPI network showed a rich interaction between DEGs. We ranked the DEGs by calculating BC values and showed the top 10 genes, followed by univariate and multifactorial analyses, which showed that GRIA2, NTRK1, SCN9A, SLC18A2, CNR1, PIK3R1, and DGKB were associated with good prognosis, and CNR1 was the most closely associated with prognosis among them. By lasso regression analysis, we constructed a four‐gene risk score formula. We drew a nomogram to use in clinical work and two newly identified genes associated with good prognosis in NB: CNR1 and GIRA2.

The case for a cancer paradox initiative.

The physicist Niels Bohr said "How wonderful that we have met with a paradox. Now we have some hope of making progress." Many paradoxical results have no obvious explanation under the standard somatic mutation theory of tumorigenesis including non-genotoxic carcinogens, foreign-body tumorigenesis, tumors lacking the inducing mutation, spontaneous regression of neuroblastoma, epithelial cancer induced by a stromal carcinogen exposure, the link between schistosomiasis and bladder cancer, and experimental reversion of cancer-like alterations to normal tissue. Alternative hypotheses of tumorigenesis provide more plausible explanations. Investigating paradoxical results in tumorigenesis using modern technology guided by various hypotheses of tumorigenesis will likely spur scientific progress and may lead to new strategies for cancer prevention.

Comparison of Stage 4 and Stage 4s Neuroblastoma Identifies Autophagy-Related Gene and LncRNA Signatures Associated With Prognosis

Background: The spontaneous regression of neuroblastoma (NB) is most prevalent and well-documented in stage 4s NB patients. However, whether autophagy plays roles in the spontaneous regression of NB is unknown.Objective: This study aimed to identify autophagy-related genes (ARGs) and autophagy-related long non-coding RNAs (lncRNAs) differentially expressed in stage 4 and stage 4s NB and to build prognostic risk signatures on the basis of the ARGs and autophagy-related lncRNAs.Methods: One RNA-sequence (RNA-Seq) dataset (TARGET NBL, n = 153) was utilized as discovery cohort, and two microarray datasets (n = 498 and n = 223) were used as validation cohorts. Differentially expressed ARGs were identified by comparing stage 4s and stage 4 NB samples. An ARG signature risk score and an autophagy-related lncRNA signature risk score were constructed. The receiver operating characteristic (ROC) curve analyses were used to evaluate the survival prediction ability of the two signatures. Gene function annotation and Gene Set Enrichment Analysis (GSEA) were performed to clarify the autophagic biological processes enriched in different risk groups.Results: Nine ARGs were integrated into the ARG signature. Patients in the high-risk group of the ARG signature had significantly poorer overall survival (OS) than patients in the low-risk group. The ROC curves analyses revealed that the ARG signature performed very well in predicting OS [5-year area under the curve (AUC) = 0.81]. Seven autophagy-related lncRNAs were integrated into the autophagy-related lncRNA signature. Patients in the high-risk group of the lncRNA signature had significantly poorer OS than patients in the low-risk group. The ROC curve analyses also revealed that the lncRNA signature performed well in predicting OS (5-year AUC = 0.77). Both the ARG signature and lncRNA signature are independent with other clinical risk factors in the multivariate Cox regression survival analyses. GSEAs revealed that autophagy-related biological processes are enriched in low-risk groups.Conclusions: Autophagy-related genes and lncRNAs are differentially expressed between stage 4 and stage 4s NB. The ARG signature and autophagy-related lncRNA signature successfully stratified NB patients into two risk groups. Autophagy-related biological processes are highly enriched in low-risk NB groups.

Identification of prognostic long noncoding RNAs associated with spontaneous regression of neuroblastoma.

BackgroundThe association between long noncoding RNAs (lncRNAs) and spontaneous regression of neuroblastoma (NB) has rarely been investigated and remains unknown.ObjectiveTo identify prognostic lncRNAs involved in the spontaneous regression of NB.MethodsDifferential expression analyses were performed between those samples with an outcome of death in stage 4 NB group and those samples with an outcome of survival in stage 4S NB group in two independent public datasets, respectively. Univariate Cox proportional hazard regression survival analysis was performed in each of the entire cohort to identify those lncRNAs significantly associated with overall survival (OS). Those lncRNAs independently associated with OS were then identified by multivariate Cox survival analysis and used to construct an lncRNA risk score.ResultsA total of 20 differentially expressed and survival‐related lncRNAs were identified sharing between the two independent cohorts. The expression of each of these 20 lncRNAs was significantly correlated with the expression of NTRK1, which is a well‐known factor involved in NB spontaneous regression. Four lncRNAs (LNC00839, FIRRE, LOC283177, and LOC101928100) were identified to be significantly associated with survival independent with each other and a four‐lncRNA signature risk score was constructed. Patients with high lncRNA signature risk score had a significantly poorer OS and event‐free survival than those with low lncRNA signature risk score. The four‐lncRNA signature has a good performance in predicting survival independent with MYCN amplification (nonamplified vs amplified), age status (<18 months vs ≥18 months), risk status (low risk vs high risk), and International Neuroblastoma Staging System (INSS) stage (INSS 1/2/3/4S vs INSS 4).ConclusionsWe identified 20 survival‐related lncRNAs that might be associated with the spontaneous regression of NB and developed a four‐lncRNA signature risk score. The four‐lncRNA signature is an independent prognostic factor for survival of NB patients.

Programmed expression of pro-apoptotic BMCC1 during apoptosis, triggered by DNA damage in neuroblastoma cells

BackgroundThe multi-functional BMCC1 (BCH motif-containing molecule at the carboxyl terminal region 1)/PRUNE2 plays a clear role in suppression of tumor activity. In the patients with neuroblastoma (NB), reduced expression of BMCC1 in primary tumor tissues was associated with poor prognosis. By contrast, enforced expression of BMCC1 as well as elevated expression of BMCC1 in response to DNA-damage promotes apoptosis by abrogating Akt-mediated survival pathways.MethodsWe addressed molecular mechanisms underlying changes in regulation of BMCC1 expression during the process of apoptosis, which was promoted by a DNA-damaging drug Cisplatin (CDDP), in NB-derived cells.ResultsElevated expression of BMCC1 was identified as an early response to DNA damage, which is accompanied by phosphorylation of ataxia telangiectasia mutated kinase (ATM) and accumulation of E2F1. Indeed, inhibition of ATM using an ATM inhibitor resulted in a decrease in expression of BMCC1 at mRNA levels. In addition, an E2F-binding sight was required for activation of BMCC1 promoter in response to DNA damage. On the other hand, knockdown of E2F1 yielded abrogated induction of BMCC1 in the cells after treatment with CDDP, suggesting that BMCC1 accumulation was caused by ATM-E2F1-dependent transcription. Finally, we demonstrated that full-length BMCC1 was proteolytically cleaved by apoptosis-activated caspase-9 during advanced stages of apoptosis in SK-N-AS cells.ConclusionsIn this study, we demonstrated the programmed expression of full-length BMCC1 in human NB cells undergoing DNA damage-induced apoptosis. The elucidation of the molecular mechanisms controlling the regulation of BMCC1 during apoptosis initiated by DNA damage provides useful information for understanding drug resistance of tumor cells and spontaneous regression of NB.

Receptor tyrosine kinase KIT can regulate the expression of genes involved in spontaneous regression of neuroblastoma

Receptor tyrosine kinase KIT can regulate the expression of genes involved in spontaneous regression of neuroblastoma

Receptor tyrosine kinase KIT may regulate expression of genes involved in spontaneous regression of neuroblastoma

Hallmark of neuroblastoma is an ability of this malignant tumor to undergo spontaneous regression or differentiation into benign tumor during any stage of the disease, but it is little known about mechanisms of these phenomena. We studied effect of receptor tyrosine kinase receptor KIT on expression of genes, which may be involved in tumor spontaneous regression. Downregulation of KIT expression by RNA interference in SH-SY5Y cells causes suppression of neurotrophin receptor NGFR expression that may promote the loss of sensibility of cells to nerve growth factors, also it causes upregulation of TrkA receptor expression which can stimulate cell differentiation or apoptosis in NGF dependent manner. Furthermore there is an upregulation of genes which stimulate malignant cell detection by immune system, such as genes of major histocompatibility complex HLA class I HLA-B and HLA-C, and interferon-γ receptors IFNGR1 and IFNGR2 genes. Thus KIT can mediate neuroblastoma cell sensibility to neurotrophins and immune system components--two factors directly contributing to spontaneous regression of neuroblastoma.

Dependence receptor UNC5D mediates nerve growth factor depletion–induced neuroblastoma regression

Spontaneous regression of neuroblastoma (NB) resembles the developmentally regulated programmed cell death (PCD) of sympathetic neurons. Regressing tumor cells express high levels of the nerve growth factor (NGF) receptors TRKA and p75NTR and are dependent on NGF for survival; however, the underlying molecular mechanism remains elusive. Here, we show that UNC5D, a dependence receptor that is directly targeted by p53 family members, is highly expressed in favorable NBs. NGF withdrawal strongly upregulated UNC5D, E2F1, and p53 in human primary favorable NBs. The induced UNC5D was cleaved by caspases 2/3, and the released intracellular fragment translocated into the nucleus and interacted with E2F1 to selectively transactivate the proapoptotic target gene. The cleavage of UNC5D and its induction of apoptosis were strongly inhibited by addition of netrin-1. Unc5d(-/-) mice consistently exhibited a significant increase in dorsal root ganglia neurons and resistance to NGF depletion-induced apoptosis in sympathetic neurons compared with wild-type cells. Our data suggest that UNC5D forms a positive feedback loop with p53 and E2F1 to promote NGF dependence-mediated PCD during NB regression.

Abstract 4842: Lysosomal-associated protein multispanning transmembrane 5 gene (LAPTM5) is associated with spontaneous regression of neuroblastomas

Abstract Background; Neuroblastoma (NB) is the most frequently occurring solid tumor in children, and shows heterogeneous clinical behavior. Favorable tumors, which are usually detected by mass screening based on increased levels of catecholamines in urine, regress spontaneously via programmed cell death (PCD) or mature through differentiation into benign ganglioneuroma (GN). In contrast, advanced-type NB tumors often grow aggressively, despite intensive chemotherapy. Understanding the molecular mechanisms of PCD during spontaneous regression in favorable NB tumors, as well as identifying genes with a pro-death role, is a matter of urgency for developing novel approaches to the treatment of advanced-type NB tumors. Key findings; We found that the expression of lysosomal associated protein multispanning transmembrane 5 (LAPTM5) was usually down-regulated due to DNA methylation in an NB cell-specific manner, but up-regulated in degenerating NB cells within locally regressing areas of favorable tumors detected by mass-screening. Experiments in vitro showed that not only a restoration of its expression but also the accumulation of LAPTM5 protein, was required to induce non-apoptotic cell death with autophagic vacuoles and lysosomal destabilization with lysosomal-membrane permeabilization (LMP) in a caspase-independent manner. While autophagy is a membrane-trafficking pathway to degrade the proteins in lysosomes, the LAPTM5-mediated lysosomal destabilization with LMP leads to an interruption of autophagic flux, resulting in the accumulation of immature autophagic vacuoles, p62/SQSTM1, and ubiqitinated proteins as substrates of autophagic degradation. In addition, ubiquitin-positive inclusion bodies appeared in degenerating NB cells. Conclusions; We propose a novel molecular mechanism for PCD with the accumulation of autophagic vacuoles due to LAPTM5-mediated lysosomal destabilization. LAPTM5-induced cell death is lysosomal cell death with impaired autophagy, not cell death by autophagy, so-called autophagic cell death. Thus LAPTM5-mediated PCD is closely associated with the spontaneous regression of NBs and opens new avenues for exploring innovative clinical interventions for this tumor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4842.

Lysosomal-Associated Protein Multispanning Transmembrane 5 Gene (LAPTM5) Is Associated with Spontaneous Regression of Neuroblastomas

BackgroundNeuroblastoma (NB) is the most frequently occurring solid tumor in children, and shows heterogeneous clinical behavior. Favorable tumors, which are usually detected by mass screening based on increased levels of catecholamines in urine, regress spontaneously via programmed cell death (PCD) or mature through differentiation into benign ganglioneuroma (GN). In contrast, advanced-type NB tumors often grow aggressively, despite intensive chemotherapy. Understanding the molecular mechanisms of PCD during spontaneous regression in favorable NB tumors, as well as identifying genes with a pro-death role, is a matter of urgency for developing novel approaches to the treatment of advanced-type NB tumors.Principal FindingsWe found that the expression of lysosomal associated protein multispanning transmembrane 5 (LAPTM5) was usually down-regulated due to DNA methylation in an NB cell-specific manner, but up-regulated in degenerating NB cells within locally regressing areas of favorable tumors detected by mass-screening. Experiments in vitro showed that not only a restoration of its expression but also the accumulation of LAPTM5 protein, was required to induce non-apoptotic cell death with autophagic vacuoles and lysosomal destabilization with lysosomal-membrane permeabilization (LMP) in a caspase-independent manner. While autophagy is a membrane-trafficking pathway to degrade the proteins in lysosomes, the LAPTM5-mediated lysosomal destabilization with LMP leads to an interruption of autophagic flux, resulting in the accumulation of immature autophagic vacuoles, p62/SQSTM1, and ubiqitinated proteins as substrates of autophagic degradation. In addition, ubiquitin-positive inclusion bodies appeared in degenerating NB cells.ConclusionsWe propose a novel molecular mechanism for PCD with the accumulation of autophagic vacuoles due to LAPTM5-mediated lysosomal destabilization. LAPTM5-induced cell death is lysosomal cell death with impaired autophagy, not cell death by autophagy, so-called autophagic cell death. Thus LAPTM5-mediated PCD is closely associated with the spontaneous regression of NBs and opens new avenues for exploring innovative clinical interventions for this tumor.

Adrenocorticotropic hormone in the aetiology and regression of neuroblastoma

Neuroblastoma is predominantly a paediatric neoplasm of the sympathetic nervous system. Despite the aggressive nature of the disease, spontaneous regression is frequently observed in infants diagnosed under the age of 12 months; especially with a specific stage referred to as stage 4s. Discovering the conditions, the elements, the mechanism and the indices behind this regression phenomenon could have therapeutic potential for prevention and cure. A review of the literature has implicated adrenocorticotropin hormone in both the aetiology and spontaneous regression of neuroblastoma. Manipulation of adrenocorticotropin hormone may offer hope for prevention and cure. Ingestible products such as retinoic acid, glycyrrhizic acid, salsolinol and ketoconazole acting in concert, could represent instrumental tools in a therapeutic manipulation process.

Past and Future of Neuroblastoma Screening in Japan

In Japan, neuroblastoma is a common malignant tumor for children and its prognosis is unfavorable. Since this tumor excretes excessive amounts of vanillylmandelic acid (VMA) and homovanillic acid (HVA), catecholamine metabolites, into urine, both are sensitive diagnostic markers for neuroblastoma. In 1974, to improve the prognosis for neuroblastoma, a mass screening program by a qualitative VMA spot test for early detection of this tumor for 6-month-old infants was started in Kyoto, Japan. Subsequently, mass screening was also conducted in other areas in Japan. In 1985, mass screening commenced as a nationwide program throughout Japan, and in 1988, screening by quantitative measurement of VMA, HVA, and creatinine was recommended by the Ministry of Health and Welfare. During the 6 years from 1984 to 1989, 468 cases of neuroblastoma were detected from among 5,052,165 infants screened by this program. Analysis of findings in the 357 cases detected by mass screening from its inception in Kyoto until December of 1988 have been presented. The survival rate for these cases was 97% (348 of 357 cases). The prognosis of tumors detected by mass screening has clearly been favorable. In addition, survival rates were improved for all cases of neuroblastoma in Kyoto after the introduction of mass screening. The increased incidence of neuroblastoma in infants and the change in age distribution of all cases of neuroblastoma following the inception of urinary screening as well as the occurrence of spontaneous regression of neuroblastoma in relation to the screening program are discussed. Future problems facing mass screening for neuroblastoma are also presented.

Commentary on screening for neuroblastoma in Japan

Professor Sawada and his Japanese colleagues are to be congratulated on their pioneering work in establishing the feasibility of whole population screening for the detection of neuroblastoma. There can be no doubt that a successful screening program would be very welcome for a disease which continues to have a most miserable outcome when it is detected in advanced stages. The review by Sawada shows (clearly that even when methods are not standardized screening is possible across the whole of Japan. We and others have followed the Japanese example and shown that whole population screening can be undertaken and that cases of neuroblastoma can be detected. Why then did a US expert committee and the International Pediatric Oncology Society recommend that there is insufficient evidence to justify the universal implementation of neuroblastoma screening and that more research is needed? The results presented by Sawada would appear to provide convincing evidence that screening is both possible, effective, worthwhile and cost effective. What then are the problems? The key problem in attempting to study the problem is the heterogeneous nature of neuroblastoma itself. It has long been known that spontaneous regression of neuroblastoma does occur. The clearest evidence for this is stage IVs neuroblastoma, for which the current recommendation for treatment is to do as little as possible. No therapy usually leads to a favorable outcome. The occasional finding of an ‘incidental’ neuroblastoma, usually asymptomatic, during radiological or other imaging procedures, the regressed ganglioneuroma of the adrenal gland found at routine autopsy of the elderly and the finding of ‘in situ’ neuroblastoma in the adrenal glands of routinely autopsied babies at a rate of 1 : 250-400 are further evidence that spontaneous regression must occur. Sawada states in his review that in the cities of Kyoto and Sapporo the detection of neuroblastoma by screening increased from 1 : 8000 to 1 : 4000 after a very sensitive HPLC screening method was

Immunobiology of neuroblastoma

Neuroblastoma remains the solid tumor of childhood with the most dismal prognosis. A more favorable survival could be expected for a number of reasons, especially if potential host anti-tumor immune activity could be augmented. On the one hand, the incidence of spontaneous regression of neuroblastoma might in part be explained by an immune mechanism; on the other, most neuroblastomas present with very large tumor burdens and the tumor expresses a paucity of self-antigens, both characteristics that hinder any effective antitumor activity. Based on historical data and today's version of host immune activity, this paper summarizes the current knowledge on both human and murine neuroblastoma and mechanisms of immunologic control.

Spontaneous Regression of Neuroblastoma: An Experimental Approach

The cytolytic activity of normal pregnancy serum was first studied on murine cancer cells and shown to be the result of a natural IgM antibody that binds to cell surfaces and activates complement. Both the classical and alternative pathways of complement are involved. It was then shown that certain human neuroblastoma cell lines, to the exclusion of other human cancers, react to the same system. It is proposed that this system may play a role in the cytolytic form of spontaneous regression of neuroblastoma.

Experimental study of the maternal effects on tumor immunity of infant mice with C1300 mouse neuroblastoma

The spontaneous regression of neuroblastoma (NB), one of the most common malignant tumors in childhood, is found to occur in 1% to 2% of patients with NB, especially in young infants. An unexpectedly favorable response to therapy is also noticed in infants suggesting the potential presence of an immune mechanism. Monoclonal C1300-S and C1300A-4 cell lines were established from polyclonal C1300 cells in our laboratory. Adult female A/J mice that had rejected 1 x 10(3) NB cells (C1300S-3) or 1 x 10(5)-10(6) NB cells (C1300A-4) were used as immunized mothers. The immunized mothers with C1300A-4 or C1300S-3 were found to have specific antibodies to C1300S-3 cells by 51Cr release assay of complement dependent cytotoxicity. Newborn mice, 24 hours after birth from immunized or nonimmunized mothers, were inoculated with 1 x 10(3) C1300S-3 NB cells. The same antibody that was assayed in the immunized mothers was detected in this offspring by the antibody-dependent cell cytotoxicity (ADCC). The tumor incidence in the offspring of the immunized mothers was found to be less than that of the offspring of the nonimmunized mothers. This study suggests that the lower tumor incidence in the offspring of immunized mothers compared with offspring of nonimmunized mothers may be attributed to their ADCC activity. Furthermore, the antibody that has the ADCC activity was proven to be immunoglobulin G by a serum absorption test using IgG absorbant. This study offers insight into the relationship between transported mother-infant immunoglobulins and on its potential control of NB.