Milica Kontic

Key epigenetic changes associated with lung cancer development: results from dense methylation array profiling.

Epigenetic alterations are a common event in lung cancer and their identification can serve to inform on the carcinogenic process and provide clinically relevant biomarkers. Using paired tumor and non-tumor lung tissues from 146 individuals from three independent populations we sought to identify common changes in DNA methylation associated with the development of non-small cell lung cancer. Pathologically normal lung tissue taken at the time of cancer resection was matched to tumorous lung tissue and together were probed for methylation using Illumina GoldenGate arrays in the discovery set (n = 47 pairs) followed by bisulfite pyrosequencing for validation sets (n = 99 pairs). For each matched pair the change in methylation at each CpG was calculated (the odds ratio), and these ratios were averaged across individuals and ranked by magnitude to identify the CpGs with the greatest change in methylation associated with tumor development. We identified the top gene-loci representing an increase in methylation (HOXA9, 10.3-fold and SOX1, 5.9-fold) and decrease in methylation (DDR1, 8.1-fold). In replication testing sets, methylation was higher in tumors for HOXA9 (p < 2.2 × 10−16) and SOX1 (p < 2.2 × 10−16) and lower for DDR1 (p < 2.2 × 10−16). The magnitude and strength of these changes were consistent across squamous cell and adenocarcinoma tumors. Our data indicate that the identified genes consistently have altered methylation in lung tumors. Our identified genes should be included in translational studies that aim to develop screening for early disease detection.

Aberrant Promoter Methylation of CDH13 and MGMT Genes is Associated With Clinicopathologic Characteristics of Primary Non–Small-Cell Lung Carcinoma

UNLABELLED Non–small-cell lung carcinoma (NSCLC) (n = 65) were analyzed for promoter methylation of RASSF1A, CDH13, MGMT, ESR1, and DAPK genes in matching lung tumors, normal lung tissue, and blood samples. Aberrant methylation in CDH13 and MGMT was associated with clinicopathologic features of NSCLC. Hypermethylation detected in primary tumors was not observed in corresponding blood samples, which rendered this an unsuitable blood-based test for NSCLC detection. INTRODUCTION Systemic methylation changes may be a diagnostic marker for tumor development or prognosis. Here, we investigate the relationship between gene methylation in lung tumors relative to normal lung tissue and whether DNA methylation changes can be detected in paired blood samples. MATERIAL AND METHODS Sixty-five patients were enrolled in a surgical case series of non-small-cell lung carcinoma at a single institution. By using bisulfite pyrosequencing, CpG methylation was quantified at 5 genes (RASSF1A, CDH13, MGMT, ESR1, and DAPK) in lung tumor, pathologically normal lung tissue, and circulating blood from enrolled cases. RESULTS The analyses of methylation in tumors compared with normal lung tissue identified higher methylation of CDH13, RASSF1A, and DAPK genes, whereas ESR1 and MGMT methylation did not differ significantly between these tissue types. We then examined whether the 3 aberrantly methylated genes could be detected in blood. The difference in methylation observed in tumors was not reflected in methylation status of matching blood samples, which indicated a low feasibility of detecting lung cancer by analyzing these genes in a blood-based test. Lastly, we probed whether tumor methylation was associated with clinical and demographic characteristics. Histology and sex were associated with methylation at the CDH13 gene, whereas, stage was associated with methylation at MGMT. CONCLUSION Our results showed higher methylation of RASSF1A, CDH13, and DAPK genes in lung tumors compared with normal lung. The lack of reflection of these methylation changes in blood samples from patients with non-small-cell lung carcinoma indicates their poor suitability for a screening test.

Intestinal Type of Lung Adenocarcinoma in Younger Adults

Intestinal type of lung adenocarcinoma (ILADC) was initially described by Tsao and Fraser in 1991. Morphology and immunophenotype of ILADC are the same as in colorectal adenocarcinoma. Rectocolonoscopy must be performed to exclude colorectal origin of adenocarcinoma. Colorectal adenocarcinoma claimed to be genetically similar to an ILADC. Patients. We describe 24- and 26-year-old patients of both genders who went under surgery because of a lung tumor mass detected on CT scan. ILADC was diagnosed on resected lung specimens. According to positivity of Cytokeratin20, CDX-2, and Villin, respectively, and negativity of Cytokeratin7, TTF-1, Napsin-A, SurfactantB, MUC-1, and MUC-2, respectively, ILADC was diagnosed. KRAS mutation was detected in tumor tissue of the male patient. Conclusion. Rectocolonoscopy is the only relevant method for distinguishing the intestinal type of lung adenocarcinoma from metastatic colorectal carcinoma because immunohistochemistry and detection of mutation status are frequently the same in both types of adenocarcinoma. More investigations are needed for further understanding of ILADC in purpose of personalized lung carcinoma therapy particularly introducing detection of mutation status, especially in younger patients.

Large Cell Lung Carcinoma with Unusual Imaging Feature, Immunophenotype and Genetic Finding

We present a case of large cell lung carcinoma in sixty-one year old male with typical lung cancer symptoms but unusual radiological presentation and immunophenotype. Tumor morphological finding related to its radiological finding was suggestive for large cell lymphoma or carcinoma, but its immunophenotype made confusion for pathological diagnosis. No p53 mutations were detected in genetic investigation. Multidisciplinar diagnostic approach to some tumors is useful for their final diagnosis.

New RB1 oncogenic mutations and intronic polymorphisms in Serbian retinoblastoma patients: genetic counseling implications

AbstractThe purpose of this work was to identify germ line RB1 mutations in 16 Serbian retinoblastoma patients for genetic counselling. Mutation analysis was carried out by PCR directed sequencing of the 27 exons. Loss of heterozygosity for two RB1 intragenic markers was also analyzed in 14 tumour samples. Five new RB1 oncogenic mutations (g.2078 del C, g.77047_48 del GC, g.78117_8 del TT, g.160797 del T, and g.64439+2 T>C) and two recurrences (R445X and Q383X) have been found in this study. In addition, four intronic variants were observed germ line in some unilateral patients. Two of these variants (g.44668-15T/G, and g.166204-8T/A) are discussed as potential oncogenic mutation candidates. The results show the relevance of studies aimed to investigate the role of intronic variants in exon splicing regulation. Such studies will help to disclose hidden retinoblastoma susceptibilities, important for accurate genetic counselling.

Diagnóstico molecular del retinoblastoma: epidemiología molecular y consejo genético

Fundamento y objetivo: El retinoblastoma, prototipo de cancer hereditario, puede causar ceguera, por enucleacion terapeutica, segundos tumores en pacientes con mutacion germinal e incluso muerte si no se trata. El diagnostico molecular de 213 pacientes a lo largo de 5 anos ha conducido a la deteccion de 106 mutaciones que se analizan desde la perspectiva de la epidemiologia molecular y consejo genetico. Pacientes y metodo: Estudio mutacional (reaccion en cadena de la polimerasa, secuenciacion y analisis de microsatelites) en pacientes con retinoblastoma procedentes de Espana, Cuba, Colombia y Serbia. Resultados: Un 45% de las mutaciones analizadas son nuevas y corresponden a mutaciones de tipo de corrimiento de pauta de lectura, cambio de aminoacido o procesado del acido ribonucleico. Todas las mutaciones sin sentido corresponden a sitios de alta mutabilidad. La tasa de deteccion de mutaciones en pacientes unilaterales esporadicos es alta (22%). En este grupo de pacientes se detecta una mayor incidencia (p = 0,018) de mutaciones de cambio de aminoacido y procesamiento. Espana y Francia muestran una incidencia mayor de mutaciones del procesado (p = 0,0003) que Alemania y el Reino Unido, paises en los que predominan las mutaciones sin sentido (p = 0,0006). Las mutaciones del procesado se asocian al fenotipo de baja penetracion y retraso en la aparicion de tumores (p = 0,018). Conclusiones: La incidencia de mutaciones germinales en pacientes unilaterales y las relaciones fenotipo/genotipo analizadas indican la necesidad del consejo genetico basado en el diagnostico molecular temprano. La mejora de las tecnicas diagnosticas, la caracterizacion funcional de mutaciones asociadas a baja penetrancia o expresividad y el estudio del transcriptoma de los tumores son objetivos necesarios para definir mejor la patogenia del retinoblastoma.

Concomitant drug- and infection-induced antineutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis with multispecific ANCA.

Objective: To report the first case of concomitant drug- and infection-induced antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) in a patient treated with propylthiouracil (PTU) and suffering from tuberculosis. Presentation and Intervention: A 28-year-old woman with PTU-treated hyperthyroidism presented with fever, purpura, pulmonary cavitations and ANCA to myeloperoxidase, bactericidal/permeability-increasing protein (BPI), proteinase-3 and elastase. Skin histopathology confirmed vasculitis. However, sputum examination revealed Mycobacterium tuberculosis. Remission was achieved after PTU withdrawal and treatment with antituberculosis drugs. Conclusion: Our case confirmed that BPI-ANCA are elevated in active tuberculosis. Multispecific ANCA were helpful for the diagnosis of concomitant PTU- and M. tuberculosis-induced AAV.

Comparison of mediastinal lymph node status and relapse pattern in clinical stage IIIA non‐small cell lung cancer patients treated with neoadjuvant chemotherapy versus upfront surgery: A single center experience

In spite of the progress made in neoadjuvant therapy for operable non small‐cell lung cancer (NSCLC), many issues remain unsolved, especially in locally advanced stage IIIA.

Angiomatosis of the thoracic wall

Angiomatosis is an extremely rare disease characterized by benign thin-walled blood vessel proliferation with asymptomatic bone destruction. This type of blood vessels proliferation with bone destruction is well known as massive osteolysis or Gorham’s disease [1, 2]. It is progressive osteolysis affecting mostly the mandible, humerus or femur. Osteolysis is usually a monostotic process. The skull is less frequently involved. Gorham’s disease becomes symptomatic in childhood or adolescence. About 200 patients with this proliferation had been described by the end of 2008 [3–5]. We describe a case of asymptomatic, incidentally detected angiomatosis affecting the thoracic wall. The patient and his parents gave consent to publish the following data. Case report

DNA Diagnosis in Oncology

It is well established that cancers are caused by the accumulation of genomic and epigenomic alterations. However, molecular changes occuring in the early stages of cancer development or in precursor lesions remain to be poorly understood.The employment of molecular characterization of tumors prior to therapy has opened the door for personalized therapy of individual patients. Such genetic alterations that are utilized to predict response to therapy are considered predictive biomarkers. The development of high throughput technologies lead to the establishment of a relatively new field of cancer genomics. These genome-wide approaches are increasingly more important in cancer diagnostic, prognosis, and treatment. Here we briefly discuss cancer transcriptome, cancer genome and epigenome analyses approches and their clinical utility. Gene expression patterns in tumors (cancer transcriptome) are analyzed by expression microarrays who can serve as a diagnostic tool which facilitates distinguishing different cancer subtypes. Another high throughput technology that may revolutionize personalized cancer therapy, lead to the development of new genetic diagnostic tests and biomarkers is next generation sequencing (NGS). NGS has an ability to fully sequence large number of genes in a single test and simultaneously detect deletions, insertions, copy number alterations, translocations and exome-wide base substitutions in cancer-related genes. NGS methods analyses DNA methylation, detection of modified histones, mapping of transcription factor occupancy, and epigenetic regulators. DNA diagnosis often trumps clinical diagnosis in oncology. This will cause the shift toward testing minute amounts of DNA and specimens including needle biopsies, circulating tumor cells, and ciruclating cell-free DNA are likely to become clinically relevant. Thus, as NGS enters clinical testing it will impact clinical decisions and cancer outcomes. Finally, improved understanding of cancer biology will lead to further development of targeted therapies and the use of combinations of targeted approaches simultaneously, increasing treatment effectiveness and supporting highly individualized patient care.