FAM120A-PTPN3 variations in neuroblastoma: Implications for poor prognosis and relapse.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e22512-e22512
Author(s):  
Lan Sun ◽  
Andres Stucky ◽  
Lingli Tu ◽  
Xuelian Chen ◽  
Jun Luo ◽  
...  
Keyword(s):  
Gene Networks ◽  
Poor Prognosis ◽  
Cohort Analysis ◽  
Neuroblastoma Cell ◽  
Gene Mutations ◽  
Current Treatment ◽  
Vitro Assay ◽  
Kaplan Meier ◽  
Mutation Distribution

e22512 Background: Neuroblastoma (NB), the most common tumor in infants, presents clinical unpredictability of relapse and treatment response. Staging and risk stratification depending on clinical examinations are often undertaken while molecular risk factors are less known. Relapse of high-risk NB remains the greatest clinical challenge. Thus, we conduct this study to investigate the molecular mechanism of NB relapse. Methods: Six patients with neuroblastoma who had both primary tumor and relapse samples were studied to identify key mutations in relapse. A cohort analysis of genetic profiles was then performed in order to find out gene mutations associated with the relapse samples. Mutation distribution and expression profiling in 127 patients from the national cancer institute database were used to identify relapse related gene networks. Kaplan-Meier analysis was performed for Overall survival (OS) on each mutation and Ingenuity pathway analysis (IPA) was consulted for their possible interactions. Gene manipulation in two neuroblastoma cell lines were used for verification. Results: We identified 40 potential mutations that associated with the relapse samples, including FAM120A and PTPN3. Mutation distribution in a larger NB population with 127 patients showed FAM120A mutation rate was 32.3% (41/127) and PTPN3 was 52.8% (67/127). Both had significant impact on OS. The median OS (mOS) with or without FAM120A mutation was 1527±214 days and 2300±139 days, respectively (p = 0.000). mOS with or without PTPN3 mutation was 1822±163 days and 2285±179 days, respectively (p = 0.037). Additionally, mOS in group lacking both mutations (42/127) was 2555±196 days, much higher than 1857±153 days, in those who had only one (62/127), or 1391±266 days, in those who had both (23/127). Furthermore, the prevalence of FAM120A mutation was at Chr.9:93543407 (61.0%, 25/41) and PTPN3 at Chr.9: 109457194 (61.2%, 41/67). Variant at Chr.9:93543407 had more significant impact on OS compared to other sites in FAM120A-mutation subgroup (p = 0.01). Expression of FAM120A was significantly down-regulated in the FAM120A-mutation subgroup compared to non-mutated (p = 0.01). IPA and in vitro assay showed these factors may have an additive effect possibly interacting through TRIM25, MYC and VIRMA. Conclusions: Our results suggest that variations in FAM120A and PTPN3 interact in neuroblastoma resulting in poor prognosis and cancer relapse, and may be a potential target for improvement of current treatment.

scholarly journals A novel in vitro assay model developed to measure both extracellular and intracellular acetylcholine levels for screening cholinergic agents

PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0258420
Author(s):  
Ryohei Tanaka-Kanegae ◽  
Koichiro Hamada
Keyword(s):  
Cell Line ◽  
Neuroblastoma Cell ◽  
Cholinergic Neurons ◽  
Mechanisms Of Action ◽  
Neuroblastoma Cell Line ◽  
Ache Inhibitor ◽  
Vitro Assay ◽  
Food Ingredients ◽  
High Doses

Background Cholinergic neurons utilize choline (Ch) to synthetize acetylcholine (ACh) and contain a high-affinity Ch transporter, Ch acetyltransferase (ChAT), ACh receptors, and acetylcholinesterase (AChE). As the depletion or malfunction of each component of the cholinergic system has been reported in patients with dementia, many studies have sought to evaluate whether treatment candidates affect each of the cholinergic components. The associated changes in the cholinergic components may be reflected by intra- or extra-cellular ACh levels, with an increase in extracellular ACh levels occurring following AChE inhibition. We hypothesized that increases in intracellular ACh levels can be more sensitively detected than those in extracellular ACh levels, thereby capturing subtle effects in the cholinergic components other than AChE. The objective of this study was to test this hypothesis. Methods We developed an in vitro model to measure both extracellular and intracellular ACh levels using the human cholinergic neuroblastoma cell line, LA-N-2, which have been reported to express Ch transporter, ChAT, muscarinic ACh receptor (mAChR), and AChE. With this model, we evaluated several drug compounds and food constituents reported to improve cholinergic function through various mechanisms. In addition, we conducted western blotting to identify the subtype of mAChR that is expressed on the cell line. Results Our cell-based assay system was capable of detecting increases in extracellular ACh levels induced by an AChE inhibitor at relatively high doses, as well as increases in intracellular ACh levels following the administration of lower AChE-inhibitor doses and an mAChR agonist. Moreover, increases in intracellular ACh levels were observed even after treatment with food constituents that have different mechanisms of action, such as Ch provision and ChAT activation. In addition, we revealed that LA-N-2 cells expressed mAChR M2. Conclusion The findings support our hypothesis and indicate that the developed assay model can broadly screen compounds from drugs to food ingredients, with varying strengths and mechanisms of action, to develop treatments for ACh-relevant phenomena, including dementia and aging-related cognitive decline.

scholarly journals NMU Is a Poor Prognostic Biomarker in Patients with Lung Adenocarcinoma

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Yan Tang ◽  
Chunsheng Hu
Keyword(s):  
Lung Adenocarcinoma ◽  
Gene Mutations ◽  
Interaction Network ◽  
Bioinformatic Analysis ◽  
Specific Gene ◽  
Aberrant Expression ◽  
Protein Protein Interaction ◽  
Web Resource ◽  
Kaplan Meier

Lung adenocarcinoma (LUAD) is the most prevalent histologic type of lung cancer, associated with a high incidence rate and substantial mortality rate worldwide. Accumulating evidence shows that the aberrant expression of neuromedin U (NMU) contributes to the initiation and progression of cancer. Herein, we explored whether NMU could be adopted as a new diagnostic and therapeutic marker in LUAD. The UALCAN and GEPIA web resources were employed to assess data on the NMU expression in LUAD. The STRING web resource was used to develop the PPI (protein-protein interaction) network of NMU, whereas Cytoscape was applied for module analysis. The Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of NMU and the interacting proteins were examined using the WebGestalt tool. Survival analysis was performed with the Kaplan-Meier plotter tool. Results revealed that the NMU expression in LUAD was significantly higher than in the nonmalignant tissues. Moreover, higher NMU levels were dramatically related to shorter overall survival, first progression survival, and postprogression survival. The specific gene mutations G45V, R143T, and F152L of NMU occurred in LUAD samples and were associated with a worse prognosis in patients. KEGG and western blot analyses demonstrated an association of NMU with the cell cycle and the cAMP signaling cascade. Bioinformatic analysis and the in vitro experiments implicated NMU as a promising prognostic signature and treatment target for LUAD.

P-Loop an T315I BCR-ABL Mutations in Chronic Myelogenous Leukemia (CML) Patients Are of Poor Prognosis, but the Y253H + E255K P-Loop Mutations Are of Even More Severe Prognosis in Chronic Phase. On the Behalf of the French Intergroup of CML (Fi-LMC Group).

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3284-3284
Author(s):  
Franck E. Nicolini ◽  
Sélim Corm ◽  
Quoc-Hung Lê ◽  
Sandrine Hayette ◽  
Nathalie Sorel ◽  
...  
Keyword(s):  
Poor Prognosis ◽  
Direct Sequencing ◽  
Chronic Phase ◽  
Progression Free Survival ◽  
Loop Group ◽  
Kinase Domain ◽  
Myelogenous Leukemia ◽  
Kaplan Meier ◽  
Group 2

Abstract In CML, the most frequent mechanism responsible for IM resistance is the onset of BCR-ABL mutations. Mutations interfering with IM binding to the ABL-kinase domain that restore the tyrosine kinase activity of BCR-ABL (T315I and P-loop mutations) are of poor prognosis. In a retrospective analysis, we analysed the features and clinical outcomes of 50 CML patients and presenting IM resistance, and harbouring 33 P-loop (1 patient had mutations) [Group 1 (G1)] and 18 T315I mutations [Group 2 (G2)] detected by direct sequencing. Twenty-eight patients were in chronic phase (CP), 12 in accelerated phase and 9 in blastic phase when IM was started, with no difference between P-loop and T315I groups. There were 23 M and 9 F, with a median age of 53 (13–74) for G1; and 14 M and 4 F, with a median age of 52 (28–70) for G2. The median duration of IM was 25 Mo (2.4–50) for G1 and 20 (0.5–145) for G2 (p=ns). The duration of IFN prior to IM was equivalent for both groups (12 Mo (0–86) for G1 vs 10 Mo (4–131) for G2, p=ns). The median interval between diagnosis and day 1 of IM was 50 Mo(0.4–128) for G1 and 20 Mo (0.5–145) (p=ns) for G2. Multivariate analysis for gender, age, prior IFN, interval diagnosis-IM start, and major cytogenetic remission achievement with IM, did not show any significant impact of these variables. Overall and Progression free survival (PFS) Kaplan Meier curves for all phases did not show any difference, however, PFS curves for CP only showed a somewhat worse survival for Y253H+E255K P-loop mutations than for other P-loop mutations or T315I (p=0.05). Figure Figure Median survival for CP was 7 Mo for Y253H+E255K P-loop mutations, vs 24 for T315I vs 36 for other P-loop mutations. In conclusion, P-loop and T315I mutations are of equivalent poor prognosis, but for CP patients, the Y253H+E255K P-loop mutations are particularly severe within the P-loop group, as suggested by in vitro data.

scholarly journals Clinical Implication of Concordant or Discordant Genomic Profiling between Primary and Matched Metastatic Tissues in Patients with Colorectal Cancer

2020 ◽  
Vol 52 (3) ◽  
pp. 764-778
Author(s):  
Jung Yoon Choi ◽  
Sunho Choi ◽  
Minhyeok Lee ◽  
Young Soo Park ◽  
Jae Sook Sung ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Machine Learning ◽  
Genetic Variants ◽  
Poor Prognosis ◽  
Gene Mutations ◽  
Genomic Profiling ◽  
Genetic Changes ◽  
Kaplan Meier ◽  
Components Analysis ◽  
Metastatic Sites

PurposeThe purpose of this study was to identify the concordant or discordant genomic profiling between primary and matched metastatic tumors in patients with colorectal cancer (CRC) and to explore the clinical implication.Materials and MethodsSurgical samples of primary and matched metastatic tissues from 158 patients (335 samples) with CRC at Korea University Anam Hospital were evaluated using the Ion AmpliSeq Cancer Hotspot Panel. We compared genetic variants and classified them as concordant, primary-specific, and metastasis-specific variants. We used a combination of principal components analysis and clustering to find genomic groups. Kaplan-Meier curves were used to appraise survival between genomic groups. We used machine learning to confirm the correlation between genetic variants and metastatic sites.ResultsA total of 282 types of deleterious non-synonymous variants were selected for analysis. Of a total of 897 variants, an average of 40% was discordant. Three genomic groups were yielded based on the genomic discrepancy patterns. Overall survival differed significantly between the genomic groups. The poorest group had the highest proportion of concordant <i>KRAS</i> G12V and additional metastasis-specific <i>SMAD4</i>. Correlation analysis between genetic variants and metastatic sites suggested that concordant <i>KRAS</i> mutations would have more disseminated metastases.ConclusionDriver gene mutations were mostly concordant; however, discordant or metastasis-specific mutations were present. Clinically, the concordant driver genetic changes with additional metastasis-specific variants can predict poor prognosis for patients with CRC.

A Simple and Inexpensive Clinical Whole Body Counter

1976 ◽  
Vol 15 (05) ◽  
pp. 248-253
Author(s):  
A. K. Basu ◽  
S. K. Guha ◽  
B. N. Tandon ◽  
M. M. Gupta ◽  
M. ML. Rehani
Keyword(s):  
The Body ◽  
Whole Body ◽  
Vitro Assay ◽  
Body Counter ◽  
Optimum Parameters ◽  
Whole Body Counter ◽  
Single Detector ◽  
Background Index ◽  
Iodide Crystal

SummaryThe conventional radioisotope scanner has been used as a whole body counter. The background index of the system is 10.9 counts per minute per ml of sodium iodide crystal. The sensitivity and derived sensitivity parameters have been evaluated and found to be suitable for clinical studies. The optimum parameters for a single detector at two positions above the lying subject have been obtained. It has been found that for the case of 131I measurement it is possible to assay a source located at any point in the body with coefficient of variation less than 5%. To add to the versatility, a fixed geometry for in-vitro counting of large samples has been obtained. The retention values obtained by the whole body counter have been found to correlate with those obtained by in-vitro assay of urine and stool after intravenous administration of 51Cr-albumin.

In Vitro Assay of Platelet Adhesiveness with Washed and Tanned Human Red Cells

1968 ◽  
Vol 20 (03/04) ◽  
pp. 384-396 ◽  
Author(s):  
G Zbinden ◽  
S Tomlin
Keyword(s):  
Platelet Adhesion ◽  
Sodium Citrate ◽  
Blood Platelets ◽  
In Vitro Assay ◽  
Red Cells ◽  
Platelet Adhesiveness ◽  
Vitro Assay ◽  
In Vitro System ◽  
Human Red Cells

SummaryAn in vitro system is described in which adhesion of blood platelets to washed and tannic acid-treated red cells was assayed quantitatively by microscopic observation. ADP, epinephrine and TAME produced a reversible increase in platelet adhesiveness which was antagonized by AMP. With Evans blue, polyanetholsulfonate, phthalanilide NSC 38280, thrombin and heparin at concentrations above 1-4 u/ml the increase was irreversible. The ADP-induced increase in adhesiveness was inhibited by sodium citrate, EDTA, AMP, ATP and N-ethylmaleimide. EDTA, AMP and the SH-blocker N-ethylmaleimide also reduced spontaneous platelet adhesion to red cells. No significant effects were observed with adenosine, phenprocoumon, 5-HT, phthalanilide NSC 57155, various estrogens, progestogens and fatty acids, acetylsalicylic acid and similarly acting agents, hydroxylamine, glucose and KCN. The method may be useful for the screening of thrombogenic and antithrombotic properties of drugs.

First Fluorescent Acetylspermidine Deacetylation Assay for HDAC10 Identifies Inhibitors of Neuroblastoma Cell Colony Growth That Increase Lysosome Accumulation

2020 ◽  
Author(s):  
Daniel Herp ◽  
Johannes Ridinger ◽  
Dina Robaa ◽  
Stephen A. Shinsky ◽  
Karin Schmidtkunz ◽  
...  
Keyword(s):  
Histone Deacetylases ◽  
High Throughput Screening ◽  
Neuroblastoma Cell ◽  
Hdac Inhibitors ◽  
Anticancer Therapy ◽  
Colony Growth ◽  
Autophagic Flux ◽  
Enzymatic Conversion ◽  
Lysine Deacetylase

Histone deacetylases (HDACs) are important epigenetic regulators involved in many diseases, esp. cancer. First HDAC inhibitors have been approved for anticancer therapy and many are in clinical trials. Among the 11 zinc-dependent HDACs, HDAC10 has received relatively little attention by drug discovery campaigns, despite its involvement e.g. in the pathogenesis of neuroblastoma. This is due in part to a lack of robust enzymatic conversion assays. In contrast to the protein lysine deacetylase and deacylase activity of the other HDAC subtypes, it has recently been shown that HDAC10 has strong preferences for deacetylation of oligoamine substrates like spermine or spermidine. Hence, it also termed a polyamine deacetylase (PDAC). Here, we present the first fluorescent enzymatic conversion assay for HDAC10 using an aminocoumarin labelled acetyl spermidine derivative to measure its PDAC activity, which is suitable for high-throughput screening. Using this assay, we identified potent inhibitors of HDAC10 mediated spermidine deacetylation in-vitro. Among those are potent inhibitors of neuroblastoma colony growth in culture that show accumulation of lysosomes, implicating disturbance of autophagic flux.

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