Frecuencia de aberraciones moleculares en pacientes cubanos con leucemia mieloide aguda / Frequency of molecular disorders in Cuban patients with acute myeloid leukemia

Introducción: En el Instituto de Hematología e Inmunología se realiza el estudio molecular de las leucemias mieloides agudas (LMA). Para las leucemias mieloides agudas no promielocíticas (LPM) se determinan cuatro biomarcadores: los genes de fusión RUNX1-RUNX1T1 y CBF(-MYH11, la duplicación interna en tándem del gen FLT3 (DIT FLT3) y la mutación A del gen NPM1 (NPM1-A). Objetivo: Determinar la frecuencia de estos cuatro biomarcadores, en pacientes cubanos con leucemias mieloides agudas primaria no promielocíticas. Métodos: Se incluyeron 91 pacientes entre niños y adultos, estudiados en el Instituto durante tres años desde el debut. A partir de ARN de sangre medular se obtuvo ADN complementario por transcripción inversa; se amplificaron los fragmentos correspondientes mediante la reacción en cadena de la polimerasa y el producto se analizó por electroforesis capilar. Resultados: El RUNX1-RUNX1T1 apareció en el 24,2 por ciento, fue más frecuente en los pacientes pediátricos y disminuyó significativamente con la edad. El CBFβ-MYH11 solo se encontró en adultos (4,8 por ciento). La NPM1-A con 41 por ciento fue mayoritaria entre los adultos. La DIT FLT3 se observó en el 21,6 por ciento y no mostró relación con la edad. NPM1-A y DIT FLT3 fueron las aberraciones con mayor presencia simultánea. Conclusiones: Por primera vez se describe la frecuencia de los cuatro biomarcadores moleculares en los pacientes cubanos con leucemias mieloides agudas primaria no promielocíticas; su comportamiento fue similar a lo descrito por otros autores, aunque se encontraron algunas particularidades(AU)

Introduction: At the Institute of Hematology and Immunology, the molecular study of acute myeloid leukemias (AML) is carried out. For nonpromyelocytic acute myeloid leukemias, four biomarkers are determined: the RUNX1-RUNX1T1 and CBF(-MYH11 fusion genes, the internal tandem duplication of the FLT3 gene (DIT FLT3), and the A mutation of the NPM1 gene (NPM1-A). Objective: To determine the frequency of these four biomarkers in Cuban patients with nonpromyelocytic primary acute myeloid leukemias. Methods: 91 patients were included, children and adults, who were studied at the Institute for three years from their disease debut. Complementary DNA was obtained from medullary blood RNA by reverse transcription. The corresponding fragments were amplified by polymerase chain reaction and the product was analyzed by capillary electrophoresis. Results: RUNX1-RUNX1T1 appeared in 24.2 percent; it was more frequent in pediatric patients and decreased significantly with age. CBFβ-MYH11 was found only in adults (4.8 percent). NPM1-A, accounting for 41 percent, represented the majority among adults. FLT3 DIT was observed in 21.6 por ciento and was not related to age. NPM1-A and DIT FLT3 were the disorders with the greatest concurrence. Conclusions: For the first time, the frequency of the four molecular biomarkers is described in Cuban patients with primary non-promyelocytic acute myeloid leukemias. Its characterization was similar to that described by other authors, although some peculiarities were found(AU)

Principales biomarcadores moleculares en la caracterización de los tumores del sistema nervioso central / Main molecular biomarkers in the characterization of central nervous system tumors

La Clasificación de Tumores del Sistema Nervioso Central de la OMS 2016 incorpora biomarcadores moleculares junto a las características histológicas clásicas, en un diagnóstico integrado, con el fin de definir distintas entidades de gliomas con la mayor precisión posible. Los estudios de perfiles moleculares en el genoma han revelado las alteraciones genéticas características y los perfiles epigenéticos asociados con diferentes tipos de gliomas. Estas características moleculares pueden usarse para refinar la clasificación del glioma, mejorar la predicción de los resultados obtenidos con los tratamientos actuales y futuros en los pacientes, y como guía de un tratamiento personalizado. Asimismo, tener una aproximación pronóstica en cada paciente. Este cambio de paradigma ha modificado la forma en que se diagnostica el glioma y sus implicancias en la práctica diaria en la indicación de los diferentes tratamientos al paciente. Aquí, sintéticamente, revisamos y destacamos los biomarcadores moleculares clínicamente relevantes. Intentamos dejar plasmado cómo los avances en la genética molecular de los gliomas pueden promover y allanar el camino hacia la medicina de precisión en neurooncología.

The Classification of Tumors of the Central Nervous System of the WHO 2016 incorporates molecular biomarkers together with the classical histological characteristics, in an integrated diagnosis, in order to define different glioma entities with the highest possible accuracy. Studies of molecular profiles in the genome have revealed characteristic genetic alterations and epigenetic profiles associated with different types of gliomas. These molecular characteristics can be used to refine the classification of gliomas, improve the prediction of the results obtained with current and future treatments in patients and as a guide for a personalized treatment. Also, have a prognostic approach in each patient. This paradigm shift has modified the way glioma is diagnosed and its implications in daily practice in the indication of different treatments to the patient. Here, synthetically, we review and highlight clinically relevant molecular biomarkers. We try to capture how advances in the molecular genetics of gliomas can promote and pave the way to precision medicine in neuro-oncology.

Role of non-coding RNAs in non-aging-related neurological disorders

Protein coding sequences represent only 2% of the human genome. Recent advances have demonstrated that a significant portion of the genome is actively transcribed as non-coding RNA molecules. These non-coding RNAs are emerging as key players in the regulation of biological processes, and act as "fine-tuners" of gene expression. Neurological disorders are caused by a wide range of genetic mutations, epigenetic and environmental factors, and the exact pathophysiology of many of these conditions is still unknown. It is currently recognized that dysregulations in the expression of non-coding RNAs are present in many neurological disorders and may be relevant in the mechanisms leading to disease. In addition, circulating non-coding RNAs are emerging as potential biomarkers with great potential impact in clinical practice. In this review, we discuss mainly the role of microRNAs and long non-coding RNAs in several neurological disorders, such as epilepsy, Huntington disease, fragile X-associated ataxia, spinocerebellar ataxias, amyotrophic lateral sclerosis (ALS), and pain. In addition, we give information about the conditions where microRNAs have demonstrated to be potential biomarkers such as in epilepsy, pain, and ALS.

Nuevas perspectivas de la citología en diagnóstico molecular del cáncer epitelial de ovario y endometrio / New perspectives of cytology in molecular diagnosis of ovarian and endometrial epithelial cancer

Resumen OBJETIVO: describir la bibliografía donde se identificaron biomarcadores moleculares en muestras de citología cervical en base líquida para detectar cáncer epitelial de endometrio y ovario. METODOLOGÍA: búsqueda electrónica en las principales bases de datos de artículos relacionados con la detección de cáncer de ovario y de endometrio con los siguientes términos: citología en base líquida, cáncer epitelial de ovario, cáncer epitelial de endometrio, biomarcadores moleculares. RESULTADOS: los cánceres epitelial de ovario y de endometrio son enfermedades que carecen de un método de tamizaje eficaz. La citología en base líquida ofrece una manera eficaz de detectar alteraciones celulares y moleculares. Mediante la asociación entre la citología en base líquida y los métodos moleculares en el aparato genital inferior es posible identificar biomarcadores específicos; esto abre la posibilidad de desarrollar métodos de tamizaje para cáncer epitelial de endometrio y ovario. CONCLUSIONES: mediante el uso de la citología en base líquida es posible detectar cáncer epitelial de endometrio y ovario.

Abstract OBJECTIVE: To describe the literature that has identified molecular biomarkers in liquid-based cervical cytology samples to detect endometrial and ovary epithelial cancer. METHODOLOGY: An electronic search was conducted in the main databases in articles related to the detection of ovarian and endometrial cancer with the following search terms: liquid-based cytology, ovarian epithelial cancer, endometrial epithelial cáncer, molecular biomarkers. RESULTS: Epithelial ovarian and endometrial cancer are diseases that lack an effective screening method. Liquid-based cytology provides an effective way to detect cellular and molecular alterations in the detection of these neoplasms. Through the association between liquid-based cytology and molecular methods it is possible to identify specific biomarkers in the lower genital tract, opening up the possibility of developing screening methods for endometrial and ovary epithelial cancer. CONCLUSIONS: It is possible the detection of endometrial cancer and, to a lesser extent of epithelial ovarian cancer by using liquid-based Pap test.

Identification of Biomarkers in Urothelium Carcinoma base on Transcriptome Sequencing / 实用医学杂志

Objective To explore the dignostic value of the candidate molecular markers in Urothelium carcinoma based on Illumina Transcriptome Sequencing. Methods Candidate genes which were screened with transcriptome sequencing were validated by using real-time fluorescent quantitative PCR in 31 pairs of tumor and normal tissues of Urothelium carcinoma patients from Affliated Hai kou Hospital Xiangya School of Medicine Central South University. Results The four candidate genes (CDH1,VEGFA,PTPRF and CLDN7) in tumor tissues were up-regulated compared with normal groups in samples of sequencing. The relative expression of VEGFA mRNA were higher than that in normal tissues. There were significant difference in two groups (P 0.05). PTPRF was closely related with the recurrence of tumor (P=0.002), and the predict sensitivity and speciality rate were 90.0%and 83.3%respectively. Conclusions CDH1 and VEGFA play important roles in the occurrence and development of bladder urothelial carcinoma, and they may be the possible biomarkers. PTPRF is expected to be a molecular reference recurrent predictor of bladder tumor.

Asociación de variantes funcionales en genes del metabolismo de la homocisteína con riesgo de trombosis venosa profunda e hiperhomocisteinemia en individuos del Sur de Chile / Association of functional variants in homocysteine metabolism genes, deep venous thrombosis risk and hyperhomocysteinemia in Southern Chilean subjects

Introducción: La Trombosis Venosa Profunda (TVP) es un importante problema de salud en la sociedad moderna. Evidencia reciente sugiere una asociación entre variantes funcionales en genes del metabolismo de la homocisteína con TVP. Sin embargo, los resultados entre poblaciones son contradictorios. En este trabajo, evaluamos la potencial asociación entre la presencia de polimorfismos en genes del metabolismo de la homocis-teína y susceptibilidad a TVP e hiperhomocisteinemia en sujetos chilenos. Métodos: Un total de 231 individuos, 77 pacientes con diagnóstico de TVP y 154 controles fueron incluidos en el estudio. Polimorfismos en los genes Metilenotetrahi-drofolato reductasa (MTHFR) y Cistationina p-sintetasa fueron genotipificados por PCR-RFLP Las concentraciones de homocisteína basal fueron cuantificadas mediante Inmunoensayo de Fluorescencia Polarizada. Resultados: La distribución genotípica y frecuencias alélicas del polimorfismo MTHFR C677T fue significativamente diferente entre pacientes y controles (p<0.01). Odds Ratio para TVP asociada al genotipo homocigoto fue 3.68 (I.C. 95 por ciento: 1.628-8.337, p<0.01). Por el contrario, la distribución genotípica de la variante CBS 844ins68 fue similar en ambos grupos (OR=1.82, I.C. 95 por ciento: 0.636-5.234, p=0.257). Además, los portadores del genotipo homocigoto MTHFR 677TT presentaron niveles más elevados de homocisteína plasmática. Conclusiones: El polimorfismo MTHFR C677T constituye un biomarcador de riesgo de TVP en población chilena, y se relaciona a mayores niveles de homo-cisteína en sujetos homocigotos. Los resultados sugieren que la detección molecular de esta variante debería ser incluida en el estudio básico de Trombofilia en nuestra población.

Background: Deep Venous Thrombosis (DVT) is an important health problem in modern society. Recent evidence suggests an association between functional variants in homocysteine metabolism genes and DVT. However, findings in different populations have been contradictory. In this work, we evaluated the potential association between the presence of polymorphisms in homocysteine metabolism genes, DVT susceptibility and hyperhomocysteinemia in Chilean subjects. Methods: A total of 231 individuals, 77 patients with diagnosis of DVT and 154 controls were included in this study. Common variants in Metylenete-trahydrofolate reductase (MTHFR) and Cistationine p-synthetase (CBS) genes were genotyped by PCR-RFLP. Basal homocysteine was quantified by Fluorescence Polarization Immunoassay. Results: Genotype distribution and allelic frequencies of MTHFR C677T polymorphism were significantly different between patients and controls. Odds Ratio for DVT associated to homozygous status was 3.68 (95 percent C.I., 1.628-8.337, p<0.01). On the other hand, the genotype distribution of the CBS 844ins68 variant was similar in both groups (OR 1.82, 95 percent C.I.: 0.636-5.234, p=0.257). In addition, the individuals carrying the MTHFR 677TT homozygous genotype exhibited higher levels of homocysteine. Conclusion: The MTHFR C677T polymorphism constituted a molecular biomarker of DVT in Chilean population, and related to higher levels of homocysteine in homozygote subjects. The results suggest that the molecular detection of this polymorphism should be included in the basic screening for thrombophilia in our population.

Differential gene expression profiles of hepatocellular carcinomas associated or not with viral infection

Chronic hepatitis B (HBV) and C (HCV) virus infections are the most important factors associated with hepatocellular carcinoma (HCC), but tumor prognosis remains poor due to the lack of diagnostic biomarkers. In order to identify novel diagnostic markers and therapeutic targets, the gene expression profile associated with viral and non-viral HCC was assessed in 9 tumor samples by oligo-microarrays. The differentially expressed genes were examined using a z-score and KEGG pathway for the search of ontological biological processes. We selected a non-redundant set of 15 genes with the lowest P value for clustering samples into three groups using the non-supervised algorithm k-means. Fisher’s linear discriminant analysis was then applied in an exhaustive search of trios of genes that could be used to build classifiers for class distinction. Different transcriptional levels of genes were identified in HCC of different etiologies and from different HCC samples. When comparing HBV-HCC vs HCV-HCC, HBV-HCC/HCV-HCC vs non-viral (NV)-HCC, HBC-HCC vs NV-HCC, and HCV-HCC vs NV-HCC of the 58 non-redundant differentially expressed genes, only 6 genes (IKBKâ, CREBBP, WNT10B, PRDX6, ITGAV, and IFNAR1) were found to be associated with hepatic carcinogenesis. By combining trios, classifiers could be generated, which correctly classified 100 percent of the samples. This expression profiling may provide a useful tool for research into the pathophysiology of HCC. A detailed understanding of how these distinct genes are involved in molecular pathways is of fundamental importance to the development of effective HCC chemoprevention and treatment.