Reclassification of Five BRCA1/2 Variants with Unknown Significance Using Complex Functional Study

PURPOSE: While BRCA1/2 genes are commonly investigated, variants of unknown significance (VUS) and variants with potential splice effect are still being detected and they represent a substantial challenge in genetic counseling and therapy. MATERIALS AND METHODS: Out of genetically tested 3,568 hereditary breast and ovarian cancer probands five, functionally not investigated variants with potential splice-modifying effect were subjected to functional characterization. Transcript-level analysis on peripheral blood-derived RNA of the carriers was performed to test aberrant splicing. The completeness of the aberrant splicing event was also studied, existence and extent of nonsense-mediated decay was even addressed. Clinical and phenotype data, pedigree and co-segregation analyses were also done. Locus-specific loss of heterozygosity (LOH) in tumor tissues was additionally tested. RESULTS: In case of the BRCA1:c.4484+4dupA and the BRCA1:c.5407-10G>A variants functional results allowed us to reclassify them from VUS into likely pathogenic category. BRCA1:c.4358-31A>C, by producing incomplete aberrant splicing, was highlighted as strong VUS, but in lack of other supporting evidence, re-categorization was not possible. The likely pathogenic assertion of previously not reported BRCA2:c.8487G>T was reinforced based on its spliceogenic property and tumor LOH, while BRCA2:c.793G>A failed to present aberrant splicing in spite of suggestive predictions, which altered its original VUS evaluation into likely benign class. CONCLUSION: We presented molecular and clinical evidence for reclassification of four out of five BRCA1/2 variants. Both up- and down-classification harbour important clinical significance. Patients carrying re-classified pathogenic variants in the future will not be dropped out from medical surveillance, preventive measures, treatment and predictive family screening in relatives at risk

Molekuláris genetikai vizsgálatok az örökletes endokrinológiai tumor szindrómák klinikai diagnosztikájában

Abstract: The common features of hereditary endocrine tumour syndromes or multiple endocrine neoplasias (MEN) are the association of various tumours of different endocrine organs in one patient or within the same family. Different types can be distinguished from among which type 1 and type 2 are the most common. The mode of inheritance is autosomal dominant, meaning that there is a 50% chance to inherit the pathogenic alteration. The pathogenic variants of genes responsible for MEN syndromes have also been identified in sporadic endocrine tumours and many cases initially referred to as sporadic have been later categorized as familiar based on genetic analysis. The main role of the molecular genetic analysis in these syndromes is to identify the pathogenic variant, then, after appropriate genetic counseling, to perform the genetic screening of first-degree relatives. Following molecular genetic analysis, the state-of-the-art clinical follow-up of the clinically healthy mutation carriers may decrease or even prevent the morbidity and mortality. Due to technological developments in recent years, the molecular genetic analysis of hereditary tumour syndromes has also been changed. Using next generation based sequencing methods in routine clinical diagnostics, the number of pathogenic genes in endocrine tumours has also increased. The present review focuses on the genetic background of hereditary endocrine tumour syndromes and the recently used molecular biological methods will also be presented. Orv Hetil. 2018; 159(7): 285?292

Fluorescence activated cell sorting followed by small RNA sequencing reveals stable microRNA expression during cell cycle progression.

BACKGROUND: Previously, drug-based synchronization procedures were used for characterizing the cell cycle dependent transcriptional program. However, these synchronization methods result in growth imbalance and alteration of the cell cycle machinery. DNA content-based fluorescence activated cell sorting (FACS) is able to sort the different cell cycle phases without perturbing the cell cycle. MiRNAs are key transcriptional regulators of the cell cycle, however, their expression dynamics during cell cycle has not been explored. METHODS: Following an optimized FACS, a complex initiative of high throughput platforms (microarray, Taqman Low Density Array, small RNA sequencing) were performed to study gene and miRNA expression profiles of cell cycle sorted human cells originating from different tissues. Validation of high throughput data was performed using quantitative real time PCR. Protein expression was detected by Western blot. Complex statistics and pathway analysis were also applied. RESULTS: Beyond confirming the previously described cell cycle transcriptional program, cell cycle dependently expressed genes showed a higher expression independently from the cell cycle phase and a lower amplitude of dynamic changes in cancer cells as compared to untransformed fibroblasts. Contrary to mRNA changes, miRNA expression was stable throughout the cell cycle. CONCLUSIONS: Cell cycle sorting is a synchronization-free method for the proper analysis of cell cycle dynamics. Altered dynamic expression of universal cell cycle genes in cancer cells reflects the transformed cell cycle machinery. Stable miRNA expression during cell cycle progression may suggest that dynamical miRNA-dependent regulation may be of less importance in short term regulations during the cell cycle

High-throughput molecular assays for inclusion in personalised oncology trials – State-of-the-art and beyond

In the last decades, the development of high-throughput molecular assays has revolutionised cancer diagnostics, paving the way for the concept of personalised cancer medicine. This progress has been driven by the introduction of such technologies through biomarker-driven oncology trials. In this review, strengths and limitations of various state-of-the-art sequencing technologies, including gene panel sequencing (DNA and RNA), whole-exome/whole-genome sequencing and whole-transcriptome sequencing, are explored, focusing on their ability to identify clinically relevant biomarkers with diagnostic, prognostic and/or predictive impact. This includes the need to assess complex biomarkers, for example microsatellite instability, tumour mutation burden and homologous recombination deficiency, to identify patients suitable for specific therapies, including immunotherapy. Furthermore, the crucial role of biomarker analysis and multidisciplinary molecular tumour boards in selecting patients for trial inclusion is discussed in relation to various trial concepts, including drug repurposing. Recognising that today's exploratory techniques will evolve into tomorrow's routine diagnostics and clinical study inclusion assays, the importance of emerging technologies for multimodal diagnostics, such as proteomics and in vivo drug sensitivity testing, is also discussed. In addition, key regulatory aspects and the importance of patient engagement in all phases of a clinical trial are described. Finally, we propose a set of recommendations for consideration when planning a new precision cancer medicine trial.imag

High-throughput molecular assays for inclusion in personalised oncology trials – State-of-the-art and beyond

In the last decades, the development of high-throughput molecular assays has revolutionised cancer diagnostics, paving the way for the concept of personalised cancer medicine. This progress has been driven by the introduction of such technologies through biomarker-driven oncology trials. In this review, strengths and limitations of various state-of-the-art sequencing technologies, including gene panel sequencing (DNA and RNA), whole-exome/whole-genome sequencing and whole-transcriptome sequencing, are explored, focusing on their ability to identify clinically relevant biomarkers with diagnostic, prognostic and/or predictive impact. This includes the need to assess complex biomarkers, for example microsatellite instability, tumour mutation burden and homologous recombination deficiency, to identify patients suitable for specific therapies, including immunotherapy. Furthermore, the crucial role of biomarker analysis and multidisciplinary molecular tumour boards in selecting patients for trial inclusion is discussed in relation to various trial concepts, including drug repurposing. Recognising that today's exploratory techniques will evolve into tomorrow's routine diagnostics and clinical study inclusion assays, the importance of emerging technologies for multimodal diagnostics, such as proteomics and in vivo drug sensitivity testing, is also discussed. In addition, key regulatory aspects and the importance of patient engagement in all phases of a clinical trial are described. Finally, we propose a set of recommendations for consideration when planning a new precision cancer medicine trial.imag

Miscellaneous. Folyóirat-referátumok. Beszámolók. KVÍZ

Folyóirat-referátumok. Endokrinológia A korai gyermekkorban elszenvedett traumás strukturális agykárosodást követően ritka a permanensen fennálló hypopituitarismus (Permanent hypopituitarism is rare after structural traumatic brain injury in early childhood) Heather, N. L., Jefferies, C., Hofman, P. L., et al. (Levelező szerző: W. S. Cutfield, Liggins Institute, University of Auckland, Private Bag 92019, 1020 Auckland, Új-Zéland): J. Clin. Endocrinol. Metab., 2012, 97, 599–604. | Kardiológia Szívmegállás terhességben: nagyon meg kell tanulni az ekkor végzendő feladatokat! (Cardiac arrest in pregnancy: lessons to be learned!) van Waning, V. H., et al. (Dept. of Intern. Med., Intensive Care Unit, Sint Franciscus Gasthuis, 3045 PM Rotterdam, Hollandia): Intensive Care Med., 2012, 38, 721. | Sportorvostan Cardiorespiratoricus fittség, alkoholfogyasztás és a metabolikus szindróma előfordulása férfiakon (Cardiorespiratory fitness, alcohol intake, and metabolic syndrome incidence in men ) Shuval, K., et al. (Levelező szerző: C. E. Barlow, The Cooper Institute, Dallas, Amerikai Egyesült Államok; e-mail: [email protected]): Med. Sci. Sports Exerc., 2012, 44, 2125–2131. | Beszámolók. A betegszervezetek szerepe az előrehaladott rákbetegek támogatásában Elhangzott 2012. október 4-én a Pécsi Orvostudományi Egyetem Ünnepi Tudományos Ülésén | Bemutatták a Ritka Betegségek Nemzeti Tervét a III. EUROTERV konferenciá

Miscellaneous. Folyóirat-referátumok

Folyóirat-referátumok. Endokrinológia Szükséges-e laboratóriumi vizsgálat phaeochromocytoma irányában natív CT-vizsgálaton alacsony denzitású mellékvese- incidentalomák esetében? (Is biochemical screening for pheochromocytoma in adrenal incidentalomas expressing low unenhanced attenuation on computed tomography necessary?) Sane, T., Schalin-Jäntti, C., Raade, M. (Department of Medicine, Division of Endocrinology, Helsinki University Central Hospital, P.O. Box 340, 00029 HUS Helsinki, Finnország; e-mail: [email protected]): J. Clin. Endocrinol. Metab., 2012, 97, 2077–2083. | Kardiológia Myocardialis infarctus utáni rövid és hosszú távú halálozás diabeteses és nem diabeteses betegek esetében, 1985–2008 (Short- and long-term mortality after myocardial infarction in patients with and without diabetes: changes from 1985 to 2008) Nauta, S. T., Deckers, J. W., Akkerhuis, K. M., et al. (Levelező szerző: Ron T. van Domburg, Thoraxcenter, Erasmus Medical Center, Rotterdam, Hollandia; e-mail: [email protected]): Diabetes Care, 2012, 35, 2043–2047. | Szívgyengeség . Az edzés csökkenti a katabolikus folyamatokat az idős krónikus szívbetegeken is (Heart failure. Exercise training attenuates MuRF-1 expression in the skeletal muscle of patients with chronic heart failure independent of age) Gielen, S., Sandri, M., et al. (Department of Internal Medicine III [Cardiology], University Hospital, Martin Luther University of Halle/ Wittenberg, Ernst-Grube Str 40, 06120 Halle, Németország; e-mail: [email protected]): Cirulation, 2012, 125, 2716–2727. | Szülészet-nőgyógyászat A férfi szexuális fejlődés a méhben: a here praenatalis leszállásának mágneses rezonanciás ábrázolása (Male sexual development in utero: testicular descent on prenatal magnetic resonance imaging) Nemec, S. F., et al. (Department of Radiology, Medical University Vienna, Waehringer Guertel 18–20, A-0190 Wien, Ausztria): Ultrasound Obstet. Gynecol. , 2011, 38, 688–694. | Traumatológia Eltévedt golyó az agyban (A stray bullet in the brain) Martucciello, G., Tripodi, R. (Department of Pediatric Science [DIPE], University of Genova, Largo Gerolamo Gaslini 5, 16147 Genova, Liguria, Olaszország; e-mail: [email protected]): Lancet, 2012, 379, e19

Miscellaneous. Levelek a szerkesztőhöz. Folyóirat-referátumok. Beszámoló

Folyóirat-referátumok. Endokrinológia Keringő mikro-RNS-ek mint potenciális diagnosztikai biomarkerek papillaris pajzsmirigy- carcinomában (Circulating microRNA profiles as potential biomarkers for diagnosis of papillary thyroid carcinoma) Yu, S., Liu, Y., Wang, J., et al. (Levelező szerző: H. Xiao, Department of Endocrinology, The Firs Affiliated Hospital, Sun Yat-sen University, 58 Zhongshan Road 2, Guangzhou, 510080, Kína): J. Clin. Endocrinol. Metab., 2012, 97, 2084–2092. | Hepatológia Az alkoholos halálozás vetülete – ébresztő felhívás (Projections of alcohol deaths – a wake-up call) Sheron, N., Hawkey, C., Gilmore, I. (Clinical Hepatology, Division of Infection, Inflammation and Immunity, Faculty of Medicine, University of Southampton, Southampton S016 6YD, Egyesült Királyság): Lancet , 2011, 377, 1297–1299. | Kardiológia Az angiotenzinreceptor-gátló neprilizin LCZ696 hatása megtartott ejekciós frakciójú szívelégtelenségben (The angiotensin receptor neprilysin inhibitor LCZ696 in heart failure with preserved ejection fraction: a phase 2 double-blind randomised controlled trial) Solomon, S. D., Zile, M., Pieske, B., et al., for the PARAMOUNT Investigators ([Dr. Solomon] Cardiovascular Division, Brigham and Women’s Hospital, Boston, MA 02115, Amerikai Egyesült Államok; e-mail: [email protected]): Lancet, 2012, 380, 1387–1395. | Kazuisztika Autoimmun kettős attak (An autoimmune double attack) Fuchs T., et al. (Levelező szerző: E. K. Wolfgang, Institute for Clinical Chemistry, University of Heidelberg, Faculty of Medicine Mannheim, Theodor-Kutzer-Ufer 1–3, D-68167 Mannheim, Németország): Lancet, 2012, 379, 1364. | Lipidológia A hypercholesterinaemia kezelésindításának alapja fiatal felnőttkorban (The rationale for initiating treatment of hypercholesterolemia in young adulthood) Steinberg, D. (Department of Medicine, School of Medicine, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093-0682, Amerikai Egyesült Államok; e-mail: dsteinberg@ucsd. edu): Curr. Atheroscler. Rep., 2013, 15, 296. | Beszámoló. A határtalan elme korlátai A Magyar Pszichiátriai Társaság XVIII. Vándorgyűlése Győr, 2013. január 23–26