@article{MTMT:3415331, title = {High and long-term von Willebrand factor expression after Sleeping Beauty transposon-mediated gene therapy in a mouse model of severe von Willebrand disease.}, url = {https://m2.mtmt.hu/api/publication/3415331}, author = {Portier, I and Vanhoorelbeke, K and Verhenne, S and Pareyn, I and Vandeputte, N and Deckmyn, H and Goldenberg, DS and Samal, HB and Singh, M and Ivics, Zoltán and Izsvak, Z and De Meyer, SF}, doi = {10.1111/jth.13938}, journal-iso = {J THROMB HAEMOST}, journal = {JOURNAL OF THROMBOSIS AND HAEMOSTASIS}, volume = {16}, unique-id = {3415331}, issn = {1538-7933}, abstract = {Essentials von Willebrand disease (VWD) is the most common inherited bleeding disorder. Gene therapy for VWD offers long-term therapy for VWD patients. Transposons efficiently integrate the large von Willebrand factor (VWF) cDNA in mice. Liver-directed transposons support sustained VWF expression with suboptimal multimerization. SUMMARY: Background Type 3 von Willebrand disease (VWD) is characterized by complete absence of von Willebrand factor (VWF). Current therapy is limited to treatment with exogenous VWF/FVIII products, which only provide a short-term solution. Gene therapy offers the potential for a long-term treatment for VWD. Objectives To develop an integrative Sleeping Beauty (SB) transposon-mediated VWF gene transfer approach in a preclinical mouse model of severe VWD. Methods We established a robust platform for sustained transgene murine VWF (mVWF) expression in the liver of Vwf(-/-) mice by combining a liver-specific promoter with a sandwich transposon design and the SB100X transposase via hydrodynamic gene delivery. Results The sandwich SB transposon was suitable to deliver the full-length mVWF cDNA (8.4 kb) and supported supra-physiological expression that remained stable for up to 1.5 years after gene transfer. The sandwich vector stayed episomal (~60 weeks) or integrated in the host genome, respectively, in the absence or presence of the transposase. Transgene integration was confirmed using carbon tetrachloride-induced liver regeneration. Analysis of integration sites by high-throughput analysis revealed random integration of the sandwich vector. Although the SB vector supported long-term expression of supra-physiological VWF levels, the bleeding phenotype was not corrected in all mice. Long-term expression of VWF by hepatocytes resulted in relatively reduced amounts of high-molecular-weight multimers, potentially limiting its hemostatic efficacy. Conclusions Although this integrative platform for VWF gene transfer is an important milestone of VWD gene therapy, cell type-specific targeting is yet to be achieved.}, year = {2018}, eissn = {1538-7836}, pages = {592-604} } @article{MTMT:2896546, title = {CIDRE: an illumination-correction method for optical microscopy}, url = {https://m2.mtmt.hu/api/publication/2896546}, author = {Smith, K and Li, YP and Piccinini, F and Csucs, G and Balazs, C and Bevilacqua, A and Horváth, Péter}, doi = {10.1038/NMETH.3323}, journal-iso = {NAT METHODS}, journal = {NATURE METHODS}, volume = {12}, unique-id = {2896546}, issn = {1548-7091}, abstract = {Uneven illumination affects every image acquired by a microscope. It is often overlooked, but it can introduce considerable bias to image measurements. The most reliable correction methods require special reference images, and retrospective alternatives do not fully model the correction process. Our approach overcomes these issues for most optical microscopy applications without the need for reference images.}, year = {2015}, eissn = {1548-7105}, pages = {404-406} } @article{MTMT:1274890, title = {Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates}, url = {https://m2.mtmt.hu/api/publication/1274890}, author = {Mátés, Lajos and Chuah, MKL and Belay, E and Jerchow, B and Manoj, N and Acosta-Sanchez, A and Grzela, DP and Schmitt, A and Becker, K and Matrai, J and Ma, L and Samara-Kuko, E and Gysemans, C and Pryputniewicz, D and Miskey, C and Fletcher, B and VandenDriessche, T and Ivics, Zoltán and Izsvák, Zsuzsa}, doi = {10.1038/ng.343}, journal-iso = {NAT GENET}, journal = {NATURE GENETICS}, volume = {41}, unique-id = {1274890}, issn = {1061-4036}, abstract = {The Sleeping Beauty (SB) transposon is a promising technology platform for gene transfer in vertebrates; however, its efficiency of gene insertion can be a bottleneck in primary cell types. A large-scale genetic screen in mammalian cells yielded a hyperactive transposase (SB100X) with B100-fold enhancement in efficiency when compared to the first-generation transposase. SB100X supported 35-50% stable gene transfer in human CD34(+) cells enriched in hematopoietic stem or progenitor cells. Transplantation of gene-marked CD34(+) cells in immunodeficient mice resulted in long-term engraftment and hematopoietic reconstitution. In addition, SB100X supported sustained (> 1 year) expression of physiological levels of factor IX upon transposition in the mouse liver in vivo. Finally, SB100X reproducibly resulted in 45% stable transgenesis frequencies by pronuclear microinjection into mouse zygotes. The newly developed transposase yields unprecedented stable gene transfer efficiencies following nonviral gene delivery that compare favorably to stable transduction efficiencies with integrating viral vectors and is expected to facilitate widespread applications in functional genomics and gene therapy.}, year = {2009}, eissn = {1546-1718}, pages = {753-761} } @article{MTMT:1046984, title = {Alpha-fetoprotein expression is a potential prognostic marker in hepatocellular carcinoma}, url = {https://m2.mtmt.hu/api/publication/1046984}, author = {Görög, Dénes and Regöly-Mérei, J and Paku, Sándor and Kopper, László and Nagy, Péter}, journal-iso = {WORLD J LGASTROENTEROL}, journal = {WORLD JOURNAL OF GASTROENTEROLOGY}, volume = {11}, unique-id = {1046984}, issn = {1007-9327}, year = {2005}, eissn = {2219-2840}, pages = {5015-5018}, orcid-numbers = {Görög, Dénes/0000-0002-4216-6282; Paku, Sándor/0000-0003-2664-7729; Kopper, László/0000-0002-4921-3678; Nagy, Péter/0000-0002-1664-9221} }