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Abstract Background: Astrocytomas are cancer tumors of the central nervous system and represent the most common type of solid tumors during human childhood. In 2016, the World Health Organization established a molecular classification system to regroup tumor entities to achieve a more accurate diagnosis and a better clinical decision-making and selection of treatment in patients with these types of tumors. Methods: We evaluated a genotyping assay for rapid and cost-effective mutation detection in astrocytomas using TaqMan probes in an asymmetric polymerase chain reaction (PCR) assay. Results: Four diffuse astrocytomas (Grade II), three anaplastic astrocytomas (Grade III), and four glioblastomas (Grade IV) were sequenced, and all of them displayed the wild-type (WT) sequence. We tried to set up this melting analysis for the genotyping of pediatric astrocytomas by identifying the specific melting temperatures of the TaqMan probes due to the presence of the WT sequences in the isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) and H3.3 histone A genes (H3F3A). We used an IDH1-TaqMan probe to identify the WT status of IDH1 in two different WT deoxyribonucleic acid (DNA) templates (pilocytic and diffuse astrocytoma) and obtained four melting temperature values ranged from 65.6 to 92.2°C. Furthermore, only four out of 29 reactions displayed amplification of the DNA template. Sanger sequencing was faster and more reliable to detect the gene status in all the sequenced samples. Conclusions: We conclude that conventional Sanger sequencing remains the gold standard for the genotyping of pediatric astrocytomas.
Resumen Introducción: Los astrocitomas son un tipo de cáncer que afecta al sistema nervioso central y representan el tumor sólido más común durante la infancia. En el año 2016, la Organización Mundial de la Salud estableció un sistema de clasificación molecular para reagrupar tumores con identidades genéticas similares y lograr un diagnóstico más preciso, lo que lleva a tomar las decisiones clínicas idóneas al elegir el tratamiento de pacientes con este tipo de tumores. Métodos: Se evaluó un protocolo que involucra el uso de sondas TaqMan en un ensayo de reacción en cadena de la polimerasa asimétrica para la detección de mutaciones en astrocitomas. Se secuenciaron cuatro astrocitomas difusos (Grado II), tres astrocitomas anaplásicos (Grado III) y cuatro glioblastomas (Grado IV). Se intentó establecer las condiciones del análisis para la genotipificación de los astrocitomas pediátricos mediante la identificación de las temperaturas de disociación específicas de las sondas TaqMan producidas por la prescencia de las secuancias WT en los genes isocitrato deshidrogenasa 1 y 2 (IDH1, IDH2) y H3.3 histona A (H3F3A). Resultados: Los astrocitomas mostraron la secuencia wild type (WT) (silvestre) de los genes. Se utilizó una sonda TaqMan IDH1 para identificar el estado de este gen en dos templados WT de DNA (astrocitoma pilocítico y difuso) y se obtuvieron cuatro valores de temperatura de disociación (65.6-92.2 °C). Solo cuatro de las 29 reacciones mostraron amplificación de DNA. La secuenciación de Sanger fue más rápida y confiable para detectar el estado de los genes en todas las muestras. Conclusiones: La secuenciación de Sanger sigue siendo la técnica más práctica para la genotipificación de astrocitomas pediátricos.
Subject(s)
Child , Humans , Astrocytoma , Brain Neoplasms , Polymerase Chain Reaction , Sequence Analysis, DNA , Genotyping Techniques , Astrocytoma/diagnosis , Astrocytoma/genetics , Brain Neoplasms/diagnosis , Histones , DNA Probes , Sequence Analysis, DNA/methods , Transition Temperature , Glioma , Isocitrate Dehydrogenase , MutationABSTRACT
Background: The World Health Organization (WHO) 2016 classification incorporated molecular subtyping in glioma, highlighting the diagnostic and prognostic significance. The study aims to determine the isocitrate dehydrogenase (IDH-1) gene, α-thalassemia/mental retardation syndrome X-linked (ATRX) gene, and tumor suppressor gene-53 (p53) mutation in glioma and their correlation with various clinical and radiological parameters.Methods: In this prospective observational study, histopathological slides of glioma (2017-2018), were analyzed for IDH-1, ATRX and p53 mutations and their correlation with various clinical and radiological parameters.Results: IDH-1 mutation was found in 48 (38.7%), ATRX loss in 38 (30.6%) and p53 mutation in 40 (32.5%) patients. The expression of IDH-1 was significantly higher (43.7%) in adults; however, no significant difference was seen with gender. Also 51.2% of patients, who presented with seizures, showed IDH-1 expression; and 27.7% of patients, who had neurological deficit also showed IDH-1 expression. IDH-1 expression was high in glioma located at insula (73.3%) and parietal lobe (71.4%); while ATRX loss was seen in glioma located at insula (80%). Intraventricular glioma characteristically lacks all three markers: IDH-1 expression, p53 overexpression and ATRX loss. IDH-1 expression and p53 overexpression was seen mainly in diffuse fibrillary astrocytoma, oligodendroglioma, anaplastic astrocytoma and glioblastoma.Conclusions: Molecular subtyping is of paramount importance in glioma management. IDH-1 mutation is commonly observed in adults and patients presenting with seizures. The duration of symptoms correlates with IDH-1 and ATRX mutations. Hypothalamic tumors lack all three mutations.
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Objective: To observe the value of MR in prediction of glioma isocitrate dehydrogenase (IDH) 1 mutation status. Methods: Nineteen-two patients with glioma were divided into IDH mutation positive group and negative group, and their imaging characteristics were retrospectively reviewed, including lesions' site, signal intensity, boundary, growth pattern, degree of enhancement and surrounding edema. Then two-class Logistic model was established. Results: There were significant differences between different grades and location of gliomas between the two groups (both P<0.05). There were no significant differences in tumor signal intensity, boundary and growth pattern (P=0.269, 0.606, 0.139). There were statistically significant difference in degree of enhancement and surrounding edema (all P<0.01). Logistic regression analysis showed that the signal uniformity (X1), boundary (X2) and degree of enhancement (X3) of gliomas were statistically significant (P=0.004, 0.037, 0.001), and the regression equation was: logit (P)=2.668+1.415X1-2.097X2-3.229X3 (χ2=41.583, P<0.001), the sensitivity of the model was 70.70%, and the specificity was 80.40%. Conclusion: MRI can be used to non-invasively predict IDH1 mutation status of gliomas before surgical operation.
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To investigate the clinical value of serum tumor marker isocitrate dehydrogenase 1(IDH1)in the diagnosis of lung cancer. The general data were collected in lung cancer patients and non-lung cancer patients.The serum level of IDH1 was detected by enzyme-linked immunosorbent assay to evaluate its clinical significance in diagnosing lung cancer. The serum IDH1 level was significantly higher in lung cancer patients than in non-lung cancer patients [(7.12±6.98)ng/ml (2.09±1.83)ng/ml,=11.540,<0.001].The serum IDH1 level in patients with adenocarcinoma or squamous cell carcinoma was significantly higher than that in patients with small cell lung cancer [(7.91±7.26)ng/ml (2.76±2.27)ng/ml, =6.345,<0.001].The sensitivity of IDH1 in detecting lung cancer,stage Ⅰ/Ⅱ lung cancer,and stage Ⅲ/Ⅳ lung cancer was 47.4%,49.1%,and 46.3%,respectively. Serum IDH1 has high sensitivities and specificities in the diagnosis and differential diagnosis of non-small cell lung cancer(squamous cell carcinoma and adenocarcinoma)and small cell lung cancer as well as the auxiliary diagnosis of stage Ⅰ and Ⅱ lung cancer.It is a valuable marker for the auxiliary diagnosis of lung cancer.
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Humans , Biomarkers, Tumor , Carcinoma, Non-Small-Cell Lung , Isocitrate Dehydrogenase , Blood , Lung NeoplasmsABSTRACT
Objective The objective of this study was to investigate the effect of systemic inflammation response index(SI-RI)on clinical prognosis of patients with glioma and its relationship with dehydrogenase 1(IDH1)mutation.Methods Eighty patients with glioma who underwent surgery in the department of Neurosurgery were collected from August 2006 to November 2015.The best clinical cutoff value for SIRI was determined using operating characteristic curve(ROC)and grouped accordingly.The Kaplan-Meier and log-rank methods were used to analyze the postoperative survival of the two groups of patients.The independent clinical prognos-tic factors were evaluated by Cox′s proportional hazards regression model.The IDH1 mutation was detected by immunohistochemistry and DNA sequencing.Results SIRI was an independent prognostic factor of glioma,and the best clinical cutoff value was 0.67 × 109/L.The median progress free survival(PFS)and overall survival(OS)of patients with low SIRI group were 46.90 months and 57.90 months,and the median PFS and OS of patients with high SIRI group were 31.78 months and 47.22 months,respectively.There was significant difference between the two groups in the median survival time of PFS and OS by log-rank method(P<0.05).Univa-riate and multivariate analysis showed that age,gender,type of surgery,WHO stage,SIRI and IDH1 mutation were the independent prognostic factors in neurostein stromal tumors.Patients with low-grade SIRI and glioma with IDH1 mutation have a better prognosis than other conditions.Conclusion SIRI is an independent prognostic factor of glioma.It is simple,convenient and reproducible,and may be used to predict the prognosis of patients with glioma.
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Objective To investigate the value of perfusion imaging in predicting the status of isocitrate dehydrogenase 1 (IDH1) in glioblastoma.Methods Retrospectively studied 30 patients with glioblastoma multiforme with wild type IDH1 (IDHw) and 30 patients with mutant IDH1 (IDH1m) in Department of Neurosurgery,Shanxi Provincial People's Hospital from September 2014 to February 2016.Ktrans and Ve within the enhancing portion of each tumor were measured by using DCE-MRI data.rCBF and rCBV within the enhancing portion of each tumor were measured by using DSC-MRI.Four parameters were represented as (x) ± s,each of the 4 parameters was compared between patients with wild type IDH1 and mutant IDH1 by using the t-test.The performance in discriminating between the two entities was evaluated by using receiver operating characteristic analysis.Results Ktrans,Ve,rCBF and rCBV inside the enhancing lesion were significantly higher in patients with wild type IDH1 than in those with mutant IDH1.There was a statistically significant difference between IDH1w group and IDH1m group of rCBF value (P ＜ 0.05).The area under the curve for Ktrans,Ve,rCBF,and rCBV inside the enhancing lesion were 0.850,0.873,0.739 and 0.772,respectively.The value of rCBF value of the Ktrans value Ve value of IDH1w was significantly higher than that of IDH1m(P ＜0.05) in the value of the Ktrans value Ve value of rCBF,the value of the AUC value in rCBV was not significantly different with the combination of the 4 parameters of the diagnostic performance (AUC =0.915).Conclusions Ktrans,Ve,rCBF and rCBV calculated from MRI are useful for predicting the IDH1 mutation status.This method is not only for the classification of brain glioblastoma diagnosis has important value,and in glioblastoma classification is of important value in the preoperative noninvasive evaluation,and it has reference significance for predicting the prognosis of patients.
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Objective To investigate the correlation between intraoperative ultrasonographic features and expression of immunohistochemical markers in patients with glioma.Methods Totally 116 patients with glioma confirmed pathologically were collected.Ultrasonic features were observed,including the main site of the tumor,maximum diameter,border,cystic degeneration,calcification,the degree of peritumoral edema and CDFI blood flow classification,and the correlation between ultrasonographic features and immunohistochemical results such as Ki-67 and isocitrate dehydrogenase 1 (IDH1) was analyzed.Results Univariate analysis showed that the border,peritumoral edema and CDFI blood flow were significantly different between negative and positive Ki-67 expression patients (P＜ 0.01).The border,cystic degeneration and peritumoral edema were distinct between negative and positive IDH1 patients (P＜0.01).Multivariate analysis showed that gliomas with clear border,high degree of peritumoral edema and rich CDFI blood flow tended to show positive Ki-67 expression,while those with vague border,low degree of peritumoral edema were frequently accompanied by positive IDH1 expression.Conclusion The border of gliomas,peritumoral edema and blood flow showed on ultrasonography may predict the expression of Ki-67 and IDH1.It is of great significance for preliminary evaluation on biological behaviors and prognosis of the tumors before surgical operation.
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OBJECTIVE: Malignant gliomas with neuronal marker expression (MGwNM) are rare and poorly characterized. Increasingly diverse types of MGwNM have been described and these reported cases underscore the dilemmas in the classification and diagnosis of those tumors. The aim of this study is to provide additional insights into MGwNM and present the clinicopathological features of 18 patients. METHODS: We reviewed the medical records of 18 patients diagnosed as MGwNM at our institute between January 2006 and December 2012. Macroscopic total resection was performed in 11 patients (61%). We evaluated the methylation status of O6-methylguanine-DNA methyltransferase (MGMT) and expression of isocitrate dehydrogenase 1 (IDH-1) in all cases, and deletions of 1p and 19q in available cases. RESULTS: The estimated median overall survival was 21.2 months. The median progression-free survival was 6.3 months. Six patients (33%) had MGMT methylation but IDH1 mutation was found in only one patient (6%). Gene analysis for 1p19q performed in nine patients revealed no deletion in six, 19q deletion only in two, and 1p deletion only in one. The extent of resection was significantly correlated with progression free survival on both univariate analysis and multivariate analysis (p=0.002 and p=0.013, respectively). CONCLUSION: In this study, the overall survival of MGwNM was not superior to glioblastoma. The extent of resection has a significant prognostic impact on progression-free survival. Further studies of the prognostic factors related to chemo-radio therapy, similar to studies with glioblastoma, are mandatory to improve survival.
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Humans , Classification , Diagnosis , Disease-Free Survival , Glioblastoma , Glioma , Isocitrate Dehydrogenase , Medical Records , Methylation , Multivariate Analysis , NeuronsABSTRACT
Objective To investigate the detection capacity of malic enzyme 1 and isocitrate dehydrogenase 1 ( Mod1&Idh1) in the laboratory animal monitoring laboratories in China in order to understand the detection capacity of la-boratories and to improve the detection level of laboratory animals’ quality.Methods Based on the program approved by CNAS, samples preparing, homogeneity test and stability test of malic enzyme 1 and isocitrate dehydrogenase 1 in the mouse kidneys were carried out.Standard operation procedure and samples with random numbers were distributed to the la-boratories.The laboratories should submit the result reports before the time limit expires.If the laboratory reports were the same with the standard results, the laboratories will receive satisfactory remark.If laboratory reports were not the same with the standard results, the laboratories will receive unsatisfactory remark.If a laboratory did not submit report, the laboratory will also receive unsatisfactory result.Results Eight laboratories out of 10 (80%) enrolled laboratories reported satisfac-tory experiment results, and two laboratories (20%) presented unsatisfactory results.Conclusions The whole detection level of laboratories in Mod1 &Idh1 is relatively high in the laboratory animals monitoring laboratories in China.It can re-flect the detection level of laboratories to conduct the laboratory capacity evaluation.
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BACKGROUND: Recent genome-wide sequencing studies have identified unexpected genetic alterations in cancer. In particular, missense mutations in isocitrate dehydrogenase-1 (IDH1) at arginine 132, mostly substituted into histidine (IDH1-R132H) were observed to frequently occur in glioma patients. METHODS: We have purified recombinant IDH1 and IDH1-R132H proteins and monitored their catalytic activities. In parallel experiments, we have attempted to find new selective IDH1-R132H chemical inhibitor(s) from a fragment-based chemical library. RESULTS: We have found that IDH1, but not IDH1-R132H, can catalyze the conversion of isocitrate into alpha-ketoglutarate (alpha-KG). In addition, we have observed that IDH1-R132H was more efficient than IDH1 in converting alpha-KG into (R)-2-hydroxyglutarate (R-2HG). Moreover, we have identified a new hit molecule, e.g., 2-(3-trifluoromethylphenyl)isothioazol-3(2H)-one as a new selective IDH1-R132H inhibitor. CONCLUSIONS: We have observed an underlying biochemical mechanism explaining how a heterozygous IDH1 mutation contributes to the generation of R-2HG and increases cellular histone H3 trimethylation levels. We have also identified a novel selective IDH1-R132H chemical hit molecule, e.g., 2-(3-trifluoromethylphenyl)isothioazol-3(2H)-one, which could be used for a future lead development against IDH1-R132H.
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Humans , Arginine , Glioma , Histidine , Histones , Mutation, MissenseABSTRACT
Isocitrate dehydrogenase 1 (IDH1), a key rate-limiting enzyme in tricarboxylic acid cycle,is found within the cytoplasm and peroxisomes.In recent years, IDH1 mutation is discovered in most gliomaand its large scale metabolites, 2-hydroxyglutarate, may be involved in tumor as potential carcinogens, and isshown to be closely related to better patients' survival as well.IDH1 mutation is anticipated to be a novel genet-ic biomarker which will guide the treatments of clinical glioma.