Stem Cell Course in the Middle East: Science Diplomacy and International Collaborations During the Arab Spring

In April 2011, an international advanced course and workshop entitled “Frontiers in Human Pluripotent Stem Cells” and an International Congress on Fertility and Genetics (http://www.fertigen.com.jo/ConferenceDetails.aspx) was held in Amman Jordan hosted by the Jordanian Society of Fertility and Genetics under the auspices of the International Cell Research Organization (ICRO), a UNESCO associated NGO. The Congress President Dr. Zaid Kilani, with Dr. Abdel Latif Abu Khadra, President of the Jordanian society for Fertility and Genetics, Dr. Rana Dajani of the Hashemite University of Jordan, and their Organizing Committee proved to be an excellent organizers and dedicated physician-scientists and, focusing on fertility, genetics and stem cells in a wide range of advanced therapeutic applications. Brilliant course participants included trainees, scientists and clinicians from the Greater Middle East. The lectures and practical sessions, presented by internationally acknowledged scientists, included overviews of recent achievements in pluripotent stem cell research, emphasizing the role of both the embryonic (ES) and induced pluripotent stem (iPS) cells. A major emphasis was placed on the clinical achievements in germ cell and umbilical cord stem cell transplantation issues, and on the potential of fast and successful prenatal and pre-implantation molecular genetics diagnostics. The organization of the stem cell course in the Holy Land especially emphasized that issues of “eternal life” and “rejuvenation” are already at hand—at least in the pluripotent stem cell research field. In the lively atmosphere of the course about 60 participants had heated discussions on the possibility and ethics of advanced prenatal diagnostics, and on regulatory issues reflecting the need of separation of clinically effective versus unapproved, unwarranted stem cell treatments. An open discussion of many ethical issues, reflecting profound differences in religion and medical tradition in the different countries, made this course exceptionally interesting for both teachers and trainees

Heterogeneous genetic background of Hungarian patients with pheochromocytoma/paraganglioma requires gene panel testing

Introduction Pheochromocytomas and paragangliomas (Pheo/PGL) are rare neuroendocrine tumours arising from the adrenal medulla or the symphathetic paraganglia, respectively. Germline mutations are present in w40% of the patients. To date, at least 16 genes have been demonstrated to be involved in the genetic background of Pheo/PGL. Prioritization in order of genes tested can be applied, but if the probability of a disease-associated germline mutation exceeds 10% the testing of all susceptibility genes is recommended. Using next generation sequencing (NGS) based methods for genetic testing of Pheo/PGL associated genes progressively becomes part of the routine diagnostics. Objective To assess the genetic background of Hungarian patients with Pheo/PGL and to develop a NGS based gene panel assay for analysis of Pheo/PGL susceptibility genes. Methods We examined 131 patients with the diagnosis of Pheo/PGL diagnosed and nursed at the 2nd Department of Medicine, Semmelweis University. The prevalence of the germline mutations of Pheo/PGL genes was determined using conventional methods. Genotype-phenotype correlations were evaluated. A gene panel covering 15 genes (RET, VHL, NF1, EPAS, EGLN1, KIF1B, SDHA, SDHB, SDHAF2, SDHC, SDHD, FH, MAX, TMEM127, MEN1) was developed and analytical sensitivity was evaluated on 36 patients with known genetic background. Library preparation was performed using SeqCapEZ capture platform with our probe design. Illumina MiSeq instrument was used for sequencing. Sequencing data were analysed with GATK workflow. Variant annotation was performed with SNPeffect. Results Germline mutations of Pheo/PGL genes were present in at 34% of the patients: 10 (7.6%) SDHB, 9 (6.9%) RET, 5 (3.8%) VHL, TMEM127, MDH2, 4 (3%) NF1, 3 (2.3%) SDHD, 2 (1.5%) SDHC and KIF1B. 5 of 10 SDHB mutation carriers developed malignant disease. Homozygous form of a MDH2 variant was associated with malignancy. Among the 10 patients with bilateral adrenal Pheo 4 RET, 2 TMEM127 and 1 VHL mutations were identified. The coverage of genes in our panel was higher than 150 reads in all regions and all known mutations were correctly identified. Discussion Our findings regarding the prevalence of germline mutations in the development of Pheo/PGL are in accordance with the literature. No founder mutation occurred in our population as we could detect mutations in 9 genes, underlining the need of novel methods for mutation analysis in everyday clinical practice. Our NGSbased gene panel performed accurately, however two recently identified genes (MDH2, GOT2) were not covered

Screening the Expression of ABCB6 in Erythrocytes Reveals an Unexpectedly High Frequency of Lan Mutations in Healthy Individuals

Lan is a high-incidence blood group antigen expressed in more than 99.9% of the population. Identification of the human ABC transporter ABCB6 as the molecular basis of Lan has opened the way for studies assessing the relation of ABCB6 function and expression to health and disease. To date, 34 ABCB6 sequence variants have been described in association with reduced ABCB6 expression based on the genotyping of stored blood showing weak or no reactivity with anti-Lan antibodies. In the present study we examined the red blood cell (RBC) surface expression of ABCB6 by quantitative flow cytometry in a cohort of 47 healthy individuals. Sequencing of the entire coding region of the ABCB6 gene in low RBC ABCB6 expressors identified a new allele (IVS9+1G>A, affecting a putative splice site at the boundary of exon 9) and two nonsynonymous SNPs listed in the SNP database (R192Q (rs150221689) and G588 S (rs145526996)). The R192Q mutation showed co-segregation with reduced RBC ABCB6 expression in a family, and we found the G588 S mutation in a compound heterozygous individual with undetectable ABCB6 expression, suggesting that both mutations result in weak or no expression of ABCB6 on RBCs. Analysis of the intracellular expression pattern in HeLa cells by confocal microscopy indicated that these mutations do not compromise overall expression or the endolysosomal localization of ABCB6. Genotyping of two large cohorts, containing 235 and 1039 unrelated volunteers, confirmed the high allele frequency of Lan-mutations. Our results suggest that genetic variants linked to lower or absent cell surface expression of ABCB6/Langereis may be more common than previously thought.This work was supported by the Lendulet Program of the Hungarian Academy of Sciences (GS), OTKA 83533 and by the Polish POIG grant 01.01.02-10-005/08 TESTOPLEK, supported by the EU through the European Regional Development Fund. Hajnalka Andrikovics is a recipient of the Janos Bolyai Research Scholarship from the Hungarian Academy of Sciences. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Dr. Camilo Toro and Dr. William Gahl of the NIH Undiagnosed Diseases Program for an affected patient specimen; that work was supported by the Intramural Research Program of the National Human Genome Research Institute and the Office of the Director of the NIH. We thank Lionel Arnaud (National Institute of Blood Transfusion (INTS), Paris, France) for helpful discussions

Genetics of Pheochromocytomas and Paragangliomas Determine the Therapeutical Approach

Pheochromocytomas and paragangliomas are the most heritable endocrine tumors. In addition to the inherited mutation other driver mutations have also been identified in tumor tissues. All these genetic alterations are clustered in distinct groups which determine the pathomechanisms. Most of these tumors are benign and their surgical removal will resolve patient management. However, 5–15% of them are malignant and therapeutical possibilities for them are limited. This review provides a brief insight about the tumorigenesis associated with pheochromocytomas/paragangliomas in order to present them as potential therapeutical targets

Synthesis and Anticancer Cytotoxicity of Azaaurones Overcoming Multidrug Resistance

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