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2. Nicht kodierende Ribonukleinsäure im kardiovaskulären System: Sonderforschungsbereich-Transregio (SFB-TRR 267)

Author

Engelhardt, S., Dimmeler, S., Heim, C., Bär, C., Böttger, T., Boon, R., Brandes, R. P., Braun, T., Dueck, A., Gagneur, J., Grote, P., Holdt, L. M., Jaé, N., Krishnan, J., Kupatt, C., Laugwitz, K.-L., Leisegang, M. S., Maegdefessel, L., Meitinger, T., Moretti, A., Müller-McNicoll, M., Sattler, M., Schnieke, A., Schulz, M., Schunkert, H., Schwalbe, H., Teupser, D., Thum, T., Weber, C., Wittig, I., and Zarnack, K.

Published
2022
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12. Nicht kodierende Ribonukleinsäure im kardiovaskulären System:Sonderforschungsbereich-Transregio (SFB-TRR 267)

Author

Engelhardt, S., Dimmeler, S., Heim, C., Bär, C., Böttger, T., Boon, R., Brandes, R. P., Braun, T., Dueck, A., Gagneur, J., Grote, P., Holdt, L. M., Jaé, N., Krishnan, J., Kupatt, C., Laugwitz, K. L., Leisegang, M. S., Maegdefessel, L., Meitinger, T., Moretti, A., Müller-McNicoll, M., Sattler, M., Schnieke, A., Schulz, M., Schunkert, H., Schwalbe, H., Teupser, D., Thum, T., Weber, C., Wittig, I., and Zarnack, K.

Abstract

The discovery of regulatory noncoding RNA molecules (ncRNA) revolutionized our previous understanding of gene expression. As shown by extensive sequencing projects from earlier times, ncRNA occurs in the human transcriptome in a previously unforeseen number and diversity, whereas mRNA coding for proteins only makes up a small proportion of the human transcriptome. In addition to structurally important ncRNA, tens of thousands of regulatory ncRNAs were identified in humans, of which microRNAs, long noncoding RNAs and the recently discovered circular RNAs are important groups. Members of this joint project have already contributed essential knowledge on the role, the mechanisms and the target structures of ncRNAs in the cardiovascular (CV) system. Furthermore, they have shown the feasibility of treatment based on the manipulation of ncRNA in cardiovascular diseases. Although a fascinating new field of research has opened up with ncRNAs, the enormous number of RNA transcripts, their different mechanisms of action and the complex methods necessary for their investigation represent major challenges. The tasks are now 1) to determine those ncRNAs which are essential for the development and homeostasis as well as the development and progression of diseases in the cardiovascular system and 2) to mechanistically understand their mode of functioning, with the aim to derive target structures and their manipulation to develop urgently needed new therapies.

Published
2022

13. A Novel UPLC-MS Metabolomic Analysis-Based Strategy to Monitor the Course and Extent of iPSC Differentiation to Hepatocytes

Author

Moreno-Torres M, Kumar M, García-Llorens G, Quintás G, Tricot T, Boon R, Tolosa L, Toprakhisar B, Chesnais F, Verfaillie C, and Castell JV

Subjects
primary human hepatocytes, metabolic pathways, iPSC differentiation, UPLC-MS, metabolomics
Abstract

Typical protocols to differentiate induced pluripotent stem cells (iPSCs) from hepatocyte-like cells (HLCs) imply complex strategies that include transfection with key hepatic transcription factors and the addition to culture media of nutrients, growth factors, and cytokines. A main constraint to evaluate the hepatic phenotype achieved arises from the way the grade of differentiation is determined. Currently, it relies on the assessment of the expression of a limited number of hepatic gene transcripts, less frequently by assessing certain hepatic metabolic functions, and rarely by the global metabolic performance of differentiated cells. We envisaged a new strategy to assess the extent of differentiation achieved, based on the analysis of the cellular metabolome along the differentiation process and its quantitative comparison with that of primary human hepatocytes (PHHs). To validate our approach, we examined the changes in the metabolome of three iPSC progenies (transfected with/without key transcription factors), cultured in three differentiation media, and compared them to PHHs. Results revealed consistent metabolome changes along differentiation and evidenced the factors that more strongly promote changes in the metabolome. The integrated dissimilarities between the PHHs and HLCs retrieved metabolomes were used as a numerical reference for quantifying the degree of iPSCs differentiation. This newly developed metabolome-analysis approach evidenced its utility in assisting us to select a cell's source, culture conditions, and differentiation media, to achieve better-differentiated HLCs.

Published
2022

15. Recent land use change

Author

Boon, R., Kay, D., Dumont, H. J., editor, Werger, M. J. A., editor, Edwards, R. W., editor, Gee, A. S., editor, and Stoner, J. H., editor

Published
1990
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17. Safety of endomyocardial biopsy in new-onset acute heart failure requiring veno-arterial extracorporeal membrane oxygenation

Author

Van Der Boon, R M A, primary, Den Dekker, W K, additional, Meuwese, C L, additional, Lorusso, R, additional, Von Der Thusen, J H, additional, Constantinescu, A C, additional, Manintveld, O C, additional, Delnoij, T S R, additional, Van Der Heijden, J J, additional, Van Mieghem, N M D A, additional, and Den Uil, C A, additional

Published
2021
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21. KBTBD13 is an actin-binding protein that modulates muscle kinetics

Author

de Winter, J M, Molenaar, J P, Yuen, M, van der Pijl, R, Shen, S, Conijn, S, van de Locht, M, Willigenburg, M, Bogaards, S J P, van Kleef, E S B, Lassche, S, Persson, Malin, Rassier, D E, Sztal, T E, Ruparelia, A A, Oorschot, V, Ramm, G, Hall, T E, Xiong, Z, Johnson, C N, Li, F, Kiss, B, Lozano-Vidal, N, Boon, R A, Marabita, M, Nogara, L, Blaauw, B, Rodenburg, R J, Kusters, B, Doorduin, J, Beggs, A H, Granzier, H, Campbell, K, Ma, W, Irving, T, Malfatti, E, Romero, N B, Bryson-Richardson, R J, van Engelen, B G M, Voermans, N C, Ottenheijm, C A C, de Winter, J M, Molenaar, J P, Yuen, M, van der Pijl, R, Shen, S, Conijn, S, van de Locht, M, Willigenburg, M, Bogaards, S J P, van Kleef, E S B, Lassche, S, Persson, Malin, Rassier, D E, Sztal, T E, Ruparelia, A A, Oorschot, V, Ramm, G, Hall, T E, Xiong, Z, Johnson, C N, Li, F, Kiss, B, Lozano-Vidal, N, Boon, R A, Marabita, M, Nogara, L, Blaauw, B, Rodenburg, R J, Kusters, B, Doorduin, J, Beggs, A H, Granzier, H, Campbell, K, Ma, W, Irving, T, Malfatti, E, Romero, N B, Bryson-Richardson, R J, van Engelen, B G M, Voermans, N C, and Ottenheijm, C A C

Abstract

The mechanisms that modulate the kinetics of muscle relaxation are critically important for muscle function. A prime example of the impact of impaired relaxation kinetics is nemaline myopathy caused by mutations in KBTBD13 (NEM6). In addition to weakness, NEM6 patients have slow muscle relaxation, compromising contractility and daily life activities. The role of KBTBD13 in muscle is unknown, and the pathomechanism underlying NEM6 is undetermined. A combination of transcranial magnetic stimulation-induced muscle relaxation, muscle fiber- and sarcomere-contractility assays, low-angle x-ray diffraction, and superresolution microscopy revealed that the impaired muscle-relaxation kinetics in NEM6 patients are caused by structural changes in the thin filament, a sarcomeric microstructure. Using homology modeling and binding and contractility assays with recombinant KBTBD13, Kbtbd13-knockout and Kbtbd13(R408c)-knockin mouse models, and a GFP-labeled Kbtbd13-transgenic zebrafish model, we discovered that KBTBD13 binds to actin - a major constituent of the thin filament - and that mutations in KBTBD13 cause structural changes impairing muscle-relaxation kinetics. We propose that this actin-based impaired relaxation is central to NEM6 pathology., Funding agencies:Dutch Foundation for Scientific Research VIDI 016.126.319Princess Beatrix Muscle Foundation W.OR17-08H2020-MSCA-RISE-2014 645648Advanced Photon Source DE-AC02-06CH11357Foundation Building Strength for Nemaline MyopathyNational Health and Medical Research Council (NHMRC) of Australia APP1121651 United States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD) NIH R01 HD075802 Muscular Dystrophy Association MDA602235 NIH National Institute of Arthritis & Musculoskeletal & Skin Diseases (NIAMS) NIH R01 AR053897 United States Department of Health & Human Services National Institutes of Health (NIH) - USA HL133359 United States Department of Energy (DOE) DE-AC02-06CH11357 NIH National Institute of General Medical Sciences (NIGMS)9 P41 GM103622 1S10OD018090-01

Published
2020
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32. SOX10 Single Transcription Factor-Based Fast and Efficient Generation of Oligodendrocytes from Human Pluripotent Stem Cells

Author

García-León, J.A. (Juan Antonio), Kumar, M. (Manoj), Boon, R. (Ruben), Chau, D. (David), One, J. (Jennifer), Wolfs, E. (Esther), Eggermont, K. (Kristel), Berckmans, P. (Pieter), Gunhanlar, N. (Nilhan), Vrij, F.M.S. (Femke), Lendemeijer, B. (Bas), Pavie, B. (Benjamin), Corthout, N. (Nikky), Kushner, S.A. (Steven A.), Dávila, J.C. (José Carlos), Lambrichts, I. (Ivo), Hu, W.-S. (Wei-Shou), Verfaillie, C.M. (Catherine M.), García-León, J.A. (Juan Antonio), Kumar, M. (Manoj), Boon, R. (Ruben), Chau, D. (David), One, J. (Jennifer), Wolfs, E. (Esther), Eggermont, K. (Kristel), Berckmans, P. (Pieter), Gunhanlar, N. (Nilhan), Vrij, F.M.S. (Femke), Lendemeijer, B. (Bas), Pavie, B. (Benjamin), Corthout, N. (Nikky), Kushner, S.A. (Steven A.), Dávila, J.C. (José Carlos), Lambrichts, I. (Ivo), Hu, W.-S. (Wei-Shou), and Verfaillie, C.M. (Catherine M.)

Abstract

Scarce access to primary samples and lack of efficient protocols to generate oligodendrocytes (OLs) from human pluripotent stem cells (hPSCs) are hampering our understanding of OL biology and the development of novel therapies. Here, we demonstrate that overexpression of the transcription factor SOX10 is sufficient to generate surface antigen O4-positive (O4+) and myelin basic protein-positive OLs from hPSCs in only 22 days, including from patients with multiple sclerosis or amyotrophic lateral sclerosis. The SOX10-induced O4+ population resembles primary human OLs at the transcriptome level and can myelinate neurons in vivo. Using in vitro OL-neuron co-cultures, myelination of neurons by OLs can also be demonstrated, which can be adapted to a high-throughput screening format to test the response of pro-myelinating drugs. In conclusion, we provide an approach to generate OLs in a very rapid and efficient manner, which can be used for disease modeling, drug discovery efforts, and potentially for therapeutic OL transplantation. In this article, García-León JA and colleagues demonstrate the generation of functional oligodendrocytes (OLs) from human pluripotent stem cells in a rapid and efficient manner by the single overexpression of SOX10. Generated OLs resemble primary OLs at the transcriptome level and can myelinate neurons both in vivo and in vitro. Neuron-OL co-cultures, adapted to high-throughput screening formats, responded to drugs affecting myelination.

Published
2018
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33. SOX10 Single Transcription Factor-Based Fast and Efficient Generation of Oligodendrocytes from Human Pluripotent Stem Cells

Author

Garcia-Leon, JA, Kumar, M, Boon, R, Chau, D, One, J, Wolfs, E, Eggermont, K, Berckmans, P, Gunhanlar, Nilhan, de Vrij, Femke, Lendemeijer, Bas, Pavie, B, Corthout, N, Kushner, Steven, Davila, JC, Lambrichts, I, Hu, WS, Verfaillie, CM, Garcia-Leon, JA, Kumar, M, Boon, R, Chau, D, One, J, Wolfs, E, Eggermont, K, Berckmans, P, Gunhanlar, Nilhan, de Vrij, Femke, Lendemeijer, Bas, Pavie, B, Corthout, N, Kushner, Steven, Davila, JC, Lambrichts, I, Hu, WS, and Verfaillie, CM

Published
2018

34. Het interviewen van getuigen

Author

Horselenberg, R., Boon, R., Odinot, G., Vredeveldt, A., Van Koppen, P.J., De Keijser, J., Jelicic, M., Criminology, A-LAB, and Empirical and Normative Studies

Published
2017

36. P3199Long non-coding RNA H19 induces abdominal aortic aneurysms

Author

Li, D Y, primary, Busch, A, additional, Jin, H H, additional, Hofmann, P, additional, Boon, R A, additional, Pelisek, J, additional, Paloschi, V, additional, Roy, J, additional, Eckstein, H H, additional, Spin, J M, additional, Tsao, P S, additional, and Maegdefessel, L, additional

Published
2018
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41. Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

Author

Ordovás, L, Boon, R, Pistoni, M, Chen, Y, Sambathkumar, R, Helsen, N, Vanhove, J, Berckmans, P, Cai, Q, Vanuytsel, K, Raitano, S, and Verfaillie, CM

Abstract

Even with the revolution of gene-targeting technologies led by CRISPR-Cas9, genetic modification of human pluripotent stem cells (hPSCs) is still time consuming. Comparative studies that use recombinant lines with transgenes integrated into safe harbor loci could benefit from approaches that use site-specific targeted recombinases, like Cre or FLPe, which are more rapid and less prone to off-target effects. Such methods have been described, although they do not significantly outperform gene targeting in most aspects. Using Zinc-finger nucleases, we previously created a master cell line in the AAVS1 locus of hPSCs that contains a GFP-Hygromycin-tk expressing cassette, flanked by heterotypic FRT sequences. Here, we describe the procedures to perform FLPe recombinase-mediated cassette exchange (RMCE) using this line. The master cell line is transfected with a RMCE donor vector, which contains a promoterless Puromycin resistance, and with FLPe recombinase. Application of both a positive (Puromycin) and negative (FIAU) selection program leads to the selection of RMCE without random integrations. RMCE generates fully characterized pluripotent polyclonal transgenic lines in 15 d with 100% efficiency. Despite the recently described limitations of the AAVS1 locus, the ease of the system paves the way for hPSC transgenesis in isogenic settings, is necessary for comparative studies, and enables semi-high-throughput genetic screens for gain/loss of function analysis that would otherwise be highly time consuming. ispartof: Journal of Visualized Experiments vol:2016 issue:117 ispartof: location:United States status: published

Published
2016

43. Young Investigator Award Session - Heart40Targeting the miRNA-106b-25 cluster as a potential regenerative therapeutic approach for myocardial injury41An allogeneic bioengineered myocardial graft limits infarct size and improves cardiac function: pre-clinical study in the porcine myocardial infarction model42Phosphoinositide 3-kinase gamma inhibition protects against anthracycline-induced cardiomyopathy by boosting cardiac autophagy43Functional screening of microRNAs identifies miR-22 as a regulator of cardiac autophagy and aging44Functional defects and molecular mechanisms of left ventricular non-compaction in nkx2.5 mutant mice45PITX2 modulates atrial membrane potential, potentiating the antiarrhythmic effects of sodium channel blockers

Author

Dirkx, E, primary, Perea Gil, I, primary, Li, MC, primary, Gupta, S K, primary, Nguyen, THM, primary, Syeda, F, primary, Dirkx, E, additional, Raso, A, additional, Braga, L, additional, Zentilin, L, additional, Zacchigna, S, additional, Giacca, M, additional, De Windt, LJ, additional, Prat-Vidal, C, additional, Galvez-Monton, C, additional, Roura, S, additional, Llucia-Valldeperas, A, additional, Soler-Botija, C, additional, Diaz-Guemes, I, additional, Crisostomo, V, additional, Sanchez-Margallo, FM, additional, Bayes-Genis, A, additional, Cimino, J, additional, De Santis, MC, additional, Pianca, N, additional, Sciarretta, S, additional, Sandri, M, additional, Zaglia, T, additional, Mongillo, M, additional, Hirsch, E, additional, Ghigo, A, additional, Bauters, C, additional, De Groote, P, additional, Foinquinos, A, additional, Boon, R, additional, Batkai, S, additional, Pinet, F, additional, Thum, T, additional, Choquet, C, additional, Kober, F, additional, Bernard, M, additional, Kelly, RG, additional, Miquerol, L, additional, Lalevee, N, additional, Holmes, A, additional, Yu, T, additional, Tull, S, additional, Kuhlmann, S, additional, Pavlovic, D, additional, Betney, D, additional, Riley, G, additional, Kucera, JP, additional, Jousset, F, additional, De Groot, J, additional, Rohr, S, additional, Brown, N, additional, Fabritz, L, additional, and Kirchhof, P, additional

Published
2016
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44. Directed Differentiation of IPS Cells to Hepatic Stellate Cells

Author

Coll, M., primary, Perea, L., additional, Boon, R., additional, Vallverdú, J., additional, Rodrigo-Torres, D., additional, Blaya, D., additional, Llopis, M., additional, Graupera, I., additional, Aguilar, B., additional, Verfaillie, C., additional, Ginès, P., additional, and Sancho-Bru, P., additional

Published
2016
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45. Highly proliferative primitive fetal liver hematopoietic stem cells are fueled by oxidative metabolic pathways

Author

KU Leuven - Inter-departmental Stem Cell Institute, Manesia, J.K., Xu, Z., Broekaert, D., Boon, R., van Vliet, A., Eelen, G., Vanwelden, T., Stegen, S., Van Gastel, N., Pascual-Montano, A., Maria-Fendt, S., Carmeliet, G., Carmeliet, P., Khurana, S., Verfaillie, C.M., KU Leuven - Inter-departmental Stem Cell Institute, Manesia, J.K., Xu, Z., Broekaert, D., Boon, R., van Vliet, A., Eelen, G., Vanwelden, T., Stegen, S., Van Gastel, N., Pascual-Montano, A., Maria-Fendt, S., Carmeliet, G., Carmeliet, P., Khurana, S., and Verfaillie, C.M.

Abstract

Hematopoietic stem cells (HSCs) in the fetal liver (FL) unlike adult bone marrow (BM) proliferate extensively, posing different metabolic demands. However, metabolic pathways responsible for the production of energy and cellular building blocks in FL HSCs have not been described. Here, we report that FL HSCs use oxygen dependent energy generating pathways significantly more than their BM counterparts. RNA-Seq analysis of E14.5 FL versus BM derived HSCs identified increased expression levels of genes involved in oxidative phosphorylation (OxPhos) and the citric acid cycle (TCA). We demonstrated that FL HSCs contain more mitochondria than BM HSCs, which resulted in increased levels of oxygen consumption and reactive oxygen species (ROS) production. Higher levels of DNA repair and antioxidant pathway gene expression may prevent ROS-mediated (geno)toxicity in FL HSCs. Thus, we here for the first time highlight the underestimated importance of oxygen dependent pathways for generating energy and building blocks in FL HSCs.

Published
2015

48. Inhibition of miR-92a improves re-endothelialization and prevents neointima formation following vascular injury

Author

Daniel, J.-M., primary, Penzkofer, D., additional, Teske, R., additional, Dutzmann, J., additional, Koch, A., additional, Bielenberg, W., additional, Bonauer, A., additional, Boon, R. A., additional, Fischer, A., additional, Bauersachs, J., additional, van Rooij, E., additional, Dimmeler, S., additional, and Sedding, D. G., additional

Published
2014
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50. Effect of body mass index on the image quality of rotational angiography without rapid pacing for planning of transcatheter aortic valve implantation: a comparison with multislice computed tomography

Author

Schultz, C. J., primary, van Mieghem, N. M., additional, van der Boon, R. M., additional, Dharampal, A. S., additional, Lauritsch, G., additional, Rossi, A., additional, Moelker, A., additional, Krestin, G., additional, van Geuns, R., additional, de Feijter, P., additional, Serruys, P. W., additional, and de Jaegere, P., additional

Published
2013
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