LUMC

 

Leiden University Medical Center

 

Contact

Pancras Hogendoorn
p.c.w.hogendoorn@lumc.nl

Anton Jan van Zonneveld
a.j.vanzonneveld@lumc.nl

Added value LUMC for hDMT:

• hiPSC derived vascular cells for microfluidic flow chips and high content screening
• hPSC derived cardiomyocytes for heart on chip formats
• mass production in bioreactors and cryopreserved supply of cardiac and vascular cells via spinout Pluriomics
• patient tissue for generating hiPSC with corresponding medical histories and informed consent for general use in screening
• hiPSC lines with various cardiac and vascular disease mutations showing disease phenotypes
• mesoderm lineage reporter hPSC lines
• technology to generate genetically engineered hPSC lines as reporters of disease phenotypes
Partner website www.lumc.nl

 

Added value hDMT for LUMC:

• opportunities for interaction and collaboration with pharma
• access to compound and drug libraries, shRNA libraries to identify disease targets
• feedback on disease models of interest to pharma
• opportunities to apply for co-funding
• access to advanced screening technology and state of the apparatus.

 

Expertise and facilities:

Expertise

Group vascular diseases (Mummery)
• Biology of pluripotent stem cells and their differentiation to the cardiac and vascular lineages
• Use of human embryonic and induced pluripotent stem cells to create cardiac and vascular(genetic) disease models
• Generation of human iPS cell lines
• Biology of heart development and cardiac disease
• Vascular diseases and the biology of endothelial and vascular smooth muscle cells
• Genetic modification of human pluripotent stem cells
• Electrophysiology of cardiac myocytes by patch clamp, voltage sensitive dyes and microelectrode arrays and their use in drug safety testing
• Calcium handling and contraction force measurements in cardiomyocytes
• Optical (optogenetic) and electrical pacing of heart cells
• Early development particularly of lateral plate, intermediate and paraxial mesoderm (heart)

Group microvascular disease (van Zonneveld, Rabelink)
• Biology of endothelial progenitor cells
• Biology and in vitro models of endothelial cell-cell and pericyte interaction
• Posttranscriptional regulation of EC and VSMC differentiation
• Access to microvascular disease-related clinical samples
• Access to patient-derived cell types suitable for iPS cell lines and the generation of in vitro models for microvascular destabilization

Group cardiac differentiation and disease (Passier)
• Heart development and disease
• Generation of human iPS cells
• Cardiac disease modeling using human iPS cells
• Human pluripotent stem cells and their differentiation to the cardiac lineage (including cardiac mesoderm, epicardial cells and cardiomyocyte subtypes such as atrial, ventricular and pacemaker cells)
• Transcriptional regulation of cardiac differentiation and disease
• Genetic modification of human pluripotent stem cells (including generation of fluorescent reporter lines)
• Electrophysiology of cardiomyocytes by patch clamp, voltage sensitive dyes and microelectrode arrays and their use in drug safety testing
• High content imaging for drug toxicity screening and cardiac biology
• Microcontact printing of cardiomyocytes
• Calcium handling and contraction force measurements in cardiomyocytes

Facilities

Mummery/Passier
• State of the art cell culture rooms, FACS, live cell imaging, electrophysiology, imaging (including optogenetics)
• iPSC core facility for generation of disease lines, development of growth and differentiation protocols based on defined culture media and extracellular matrix proteins
• Availability of human fetal tissue for research

Van Zonneveld/Rabelink
• A pericyte stabilized microfluidic model for microvascular destabilization (with Hankemeijer/Mimetas)
• Access to large collections of characterized clinical samples of patients relevant for the study of microvascular disease

 

Ongoing projects:

Mummery/Passier
• Endothelial and smooth muscle cells in microfluidic chips (with Mimetas, UT): Muller Foundation; Plurimes FP7 EU)
• Cardiomyocytes in microfluidic chips (with Mimetas)
• Cytostretch (with TU Delft)
• Pole technology (with Thomas Schmidt): FOM
• Methods for cardiomyocyte maturation (cyclic stretch, hormones, pacing, optogenetics): Crack-It (phase 1, phase 2); ERC

Van Zonneveld/Rabelink
• Microvascularity in OrganoPlates: A microvascular model is developed in different tissue contexts to study the influence of microvasculature on organ damage using metabolomics techniques (with LACDR & MIMETAS).
• Impact of diabetes and renal failure on microvascular de-stabilization (NSN, NIRM)
•. Gender specific mechanisms in microvascular dysfunction in HFPEF (CVON, NHS)
• Microvascular rarefaction in Atrial Fibrillation and Kidney transplantation (NHS, BMS)

 

Publications:

• Davis, R.P. Casini, S., van den Berg, C., Hoekstra, M., Remme, C., Dambrot, C., Salvatori, D., Ward-van Oostwaard, D., Wilde, A.A., Bezzina, C.R., Verkerk, A., Freund, C., Mummery, C.L. Cardiomyocytes derived from pluripotent stem cells recapitulate electrophysiologicalcharacteristics of an overlap syndrome of cardiac sodium channel disease. Circulation, 2012 125(25):3079-91.
• Braam, S.R., Tertoolen, L., Casini, S., Matsa, E., Lu, H.R., Teisman, A., Passier, R., Denning, C., Gallacher, D.J., Towart, R., Mummery, C.L. Repolarization reserve determines drug responses in human pluripotent stem cell derived cardiomyocytes Stem Cell Res. 2013 10(1):48-56.
• Bellin, M., Davis, R.P., Casini, S., et al Isogenic human pluripotent stem cell pairs reveal the role of a KCNH2 mutation in long-QT syndrome. EMBO J 2013 32, 3161–3175.
• Orlova, V.V., Drabsch, Y., Freund, C., Petrus Reurer, S., van den Hil, F.E., Muenthaisong, S., ten Dijke, P., Mummery. C.L. Functionality of endothelial cells and pericytes from human pluripotent stem cells demonstrated in cultured vascular plexus and zebrafish xenografts,Arterioscler. Thromb. Vasc. Biol. 2014 34,177–186.
• Orlova, V.V., van den Hil, F.E., Petrus-Reurer, S., Drabsch, Y., ten Dijke, P., Mummery, C.L. Endothelial Cells and Pericytes from human Pluripotent Stem Cells: methods for efficient generation, expansion and examination of functional competence. Nature Protocols 2014,9:1514-1531.
• Den Hartogh, S.C., Schreurs, C., Monshouwer-Kloots, J.J., Davis, R.P., Elliott, D.A., Mummery, C.L., Passier, R. Dual reporter MESP1mCherry/w-NKX2-5eGFP/w hESCs enable tracking of early human cardiac differentiation. Stem Cells. 2014 Sep 3. doi: 10.1002/stem.1842.
• Bijkerk, R., van Solingen, C., de Boer, H.C., van der Pol, P., Khairoun, M., de Bruin, R.G., van Oeveren-Rietdijk, A.M., Lievers, E., Schlagwein, N., van Gijlswijk, D.J., Roeten, M.K., Neshati, Z., de Vries, A.A., Rodijk, M., Pike-Overzet, K., van den Berg, Y.W., van der Veer, E.P., Versteeg, H.H., Reinders, M.E., Staal, F.J., van Kooten, C., Rabelink, T.J., van Zonneveld, A.J. Hematopoietic microrna-126 protects against renal ischemia/reperfusion injury by promoting vascular integrity. Journal of the American Society of Nephrology : JASN. 2014;25:1710-1722.
• van der Veer, E.P., de Bruin, R.G., Kraaijeveld, A.O., de Vries, M.R., Bot, I., Pera, T., Segers, F.M., Trompet, S., van Gils, J.M., Roeten, M.K., Beckers, C.M., van Santbrink, P.J., Janssen, A., van Solingen, C., Swildens, J., de Boer, H.C., Peters, E.A., Bijkerk, R., Rousch, M., Doop, M., Kuiper, J., Schalij, M.J., van der Wal, A.C., Richard, S., van Berkel, T.J., Pickering, J.G., Hiemstra, P.S., Goumans, M.J., Rabelink, T.J., de Vries, A.A., Quax ,P.H., Jukema, J.W., Biessen, E.A., van Zonneveld, A.J. Quaking, an rna-binding protein, is a critical regulator of vascular smooth muscle cell phenotype. Circulation research. 2013;113:1065-1075.
• de Boer, H.C., Hovens, M.M., van Oeveren-Rietdijk, A.M., Snoep, J.D., de Koning, E.J.,Tamsma, J.T., Huisman, M.V., Rabelink, A.J., van Zonneveld, A.J. Human cd34+/kdr+ cells are generated from circulating cd34+ cells after immobilization on activated platelets. Arteriosclerosis, thrombosis, and vascular biology. 2011;31:408-415.
• Rabelink, T.J., de Boer, H.C., van Zonneveld, A.J. Endothelial activation and circulating markers of endothelial activation in kidney disease. Nature reviews. Nephrology. 2010;6:404-414.

 

Relevant patents:

Development of methods for efficient production of endothelial cells and smooth muscle cells from human Pluripotent Stem Cells (hPSC) (ATVB 2013 en Nat. Protocols, 2014), and a 2D culture system that mimics tumor blood vessels (ATVB 2013). Patent filed.