As average life expectancy increases, so does the prevalence of cardiovascular, neurological and neurodegenerative disease. This pending epidemic is of great concern because of the lack of any effective treatments. Preventing or delaying the onset of symptoms is thought to be a better strategy than attempting to reverse them once they occur, but doing this requires more insight into the molecular and genetic nature of the diseases. Since vascular malfunction is a common feature for all of these conditions, it is essential to study how this occurs in humans.

Human vessels on microfluidic chips

Animal models and primary human vascular cells grown on plastic tissue culture dishes have so far failed to capture human vascular disease effectively and have poor predictive value for drug responses. hDMT is therefore pioneering modeling of human vessels on microfluidic chips based on induced Pluripotent Stem cells (iPS). The resulting devices contain multicellular tubular structures resembling human vessels through which fluid can flow.  These can be used for vascular disease research, drug discovery and screening of compounds that affect vessel integrity.

Because vessels are an integral part of all tissues in the human body and play an important role in inflammation and metastasis of cancer, hDMT has given high priority to this research program.

Program coordinators

Chair
Valeria Orlova
LUMC

Vice-chair
Antoinette Maassen van den Brink
Erasmus MC

Upcoming meeting

  • 12 June (10-16h, Amsterdam UMC)
  • 27 November (12:30-17:30h, LUMC)

Previous meetings

  • 7 December 2023 (13-17h LUMC)
  • 8 June 2023 (Online)
  • 15 December 2022
  • 20 May 2022
  • 3 June 2021, Online (13-16h)
  • 8 December 2020, Online
  • 10 June 2020, Online
  • 13 June 2019, Leiden (host LUMC)

Making human vessels on a chip

The first step in creating human vessel mimics is to make iPS cells from skin, blood or cells in urine. In some cases these will be from patients with genetic diseases that affect vessels. The iPS cells can be turned into endothelial cells and pericytes (or mural cells) that are the cellular components of the vasculature.  By carefully making different types of endothelial- and mural cells, it is becoming possible to create arteries, veins, capillaries or lymph vessels. These cells are then introduced into microfluidic devices where under conditions of flow they organize themselves into three-dimensional tubes, with an inner lining of endothelial cells and an outer layer of smooth muscle cells, just as vessels in the body. It is then possible, for example, to add immune cells to the circulating fluid in order to study the effects of inflammation on vascular integrity or interrupt flow to resemble thrombosis.

Future outlook

The long-term vision of hDMT is to create complex organ systems on chips, with vessel mimics connecting the organ tissues. To facilitate drug testing it will be possible, for example, to deliver liver cells to one culture chamber and kidney or brain cells in another. Any drug added could then be metabolized in the liver compartment and the metabolic products, which are often the major cause of toxicity, could flow to the target tissue in the adjacent chamber. More tissues could be added, ultimately leading to a ‘body on chip’ – a powerful approach for studying the interaction between the organs and the effects of drugs throughout the body.

Customized vessels on chip

In the distant future, it is conceivable that many individuals will have their own iPS cells stored in banks, much like blood is stored today. Using these cells, it may be possible to make customized vessels on chip, allowing analysis of drug responses on an individual basis for personalized medicine. These ‘patients on chip’ could facilitate identification of the most effective drug candidates for a vascular disease in a given patient, saving time, money and discomfort. It may also become possible to predict health problems for people with genetic predisposition for certain vascular diseases, in some cases presenting opportunities for lifestyle adjustments that could delay onset of the ailment in question.

However, much research remains to be done before this vision turns into reality. Partners in the hDMT Vessel on Chip consortium nevertheless intend to contribute to making it happen.

Interdisciplinary research

Vessels on Chip is an interdisciplinary program to which each partner contributes depending on their own ongoing research and clinical interest. Leiden University Medical Center provides (anonymized) patient samples and corresponding medical histories and turns the patient’s cells into iPS cells that can be made to form any type of cell in the human body. This includes the vascular endothelial- and mural cells that make up vessels. Each type of blood- (and lymph) vessel, arterial, venous or capillary can be identified and characterized by single cell mRNA sequencing technology at the Hubrecht Institute.

Novel methods

Groups at the Technical Universities are experts in microfluidics and microfabrication, and already have novel methods for measuring vascular integrity and dysfunction operational. The Leiden Academic Centre for Drug Research contributes expertise in metabolomics and mass spectrometry to assess secreted biomarkers in minute volumes of fluid flowing through the vessels on chip. Working closely with biotech companies, such as Pluriomics (for mass cell production), Mimetas (device development and test) and Galapagos (drug discovery), hDMT aims to create models and bioassays with quantitative readouts for multiple chronic and acute vascular diseases. High throughput manufacturing is facilitated by cooperation with Philips.  

Vascular disease models

One of the biggest health challenges in western society is diabetes, a chronic disease that is associated with serious vascular complications, especially in the kidneys and eyes. Renal failure is the leading cause of death in diabetics and diabetes is the major cause of non-age associated blindness. As there is little known about the underlying mechanisms of nephropathy and retinopathy, and even less about possible treatments, there is an urgent need for diabetic vasculitis models. Mimics of the (highly specialized) retinal and glomerulus vascular beds will enable us to get an insight into the causes and development of these pathologies, potentially leading to better therapies.

 

Realistic model

hDMT also investigates the vascular contribution to neurological and neurodegenerative diseases. Patients with blood vessel malformations in the brain are at risk of developing neurological disorders that resemble dementia, possibly caused by the breakdown of the blood-brain barrier. Understanding the mechanisms of neurovascular dysfunction and the mechanisms that regulate the blood-brain barrier, is of high priority for prevention of these diseases and for identification of potential drug targets. A realistic model of the blood-brain barrier could also provide new insights into how to deliver drugs to the brain across this protective shield; this is vital for treating diseases like brain cancer or epilepsy.  

Cancer cells

A third focus area of the vessel on chip program is tumor biology, since tumors need blood vessels to develop and spread. Combining human vessels with cancer cells can provide new information on how to promote the formation of blood vessels in and around a tumor for drug delivery and block this before the blood starts nourishing the tumor.  

The use of human iPS cells in hDMT’s vascular disease models offers unprecedented opportunities for mimicking genetic diseases in vitro, since the genome of the patient is captured in the derivative vasculature in culture. LUMC has already successfully applied this technology to a number of hereditary vascular disorders. 

News

Thesis Heleen Middelkamp about microfluidic vessels-on-chips for vascular disease modelling

On Friday September 8th 2023 Heleen Middelkamp successfully defended her PhD thesis entitled ‘Microfluidic vessels-on-chips for vascular disease modelling’ at the University of Twente.

Read more

Publications

Andreas Pollet and Jaap den Toonder (2020). Review: Recapitulating the vasculature using Organ-on-Chip technology. Bioengineering, 7, 17.

Andreas Pollet, Erik Homburg, Ruth Cardinaels and Jaap den Toonder (2020). 3D Sugar Printing of Networks Mimicking the Vasculature. Micromachines, 11, 4.3

de Graaf MNS, Cochrane A, van den Hil FE, Buijsman W, van der Meer AD, van den Berg A, Mummery CL, Orlova VV. (2019) Scalable microphysiological system to model three-dimensional blood vessels. APL Bioeng. Jun 21; 3(2):026105.

Cao X, Yakala GK, van den Hil FE, Cochrane A, Mummery CL, Orlova VV. (2019) Differentiation and Functional Comparison of Monocytes and Macrophages from hiPSCs with Peripheral Blood Derivatives. Stem Cell Reports. Jun 11; 12(6):1282-1297.

 

Oleh V. Halaidych, Amy Cochrane, Francijna E. van den Hil, Christine L. Mummery and Valeria V. Orlova, Quantitative Analysis of Intracellular Ca2+Release and Contraction in hiPSC-Derived Vascular Smooth Muscle Cells. Stem Cell Reports. Published: March 7 (2019) DOI:https://doi.org/10.1016/j.stemcr.2019.02.003

Oleh V. Halaidych, Francijna E. van den Hil, Christine L. Mummery and Valeria V. Orlova, Microfluidic Assay for the Assessment of Leukocyte Adhesion to Human Induced Pluripotent Stem Cell-derived Endothelial Cells (hiPSC-ECs). J Vis Exp. (2018) Nov 26; (141). doi: 10.3791/58678.

1. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T. et al. (2007). Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131: 861-872.

2. Bellin, M., Marchetto, M.C., Gage, F.H.,  Mummery, C.L. Induced pluripotent stem cells: the new patient? Nat. Rev. Mol. Cell Biol. 13, 713–726 (2012).    

3. Antonetti, D. A., Klein, R. & Gardner, T. W. Diabetic retinopathy. N. Engl. J. Med. 366, 1227–1239 (2012).

4. Rask-Madsen, C. & King, G. L. Vascular Complications of Diabetes: Mechanisms of Injury and Protective Factors. Cell Metab. 17, 20–33 (2013).

5. Eremina, V. et al. VEGF inhibition and renal thrombotic microangiopathy. N. Engl. J. Med. 358, 1129–1136 (2008).

6. Gorelick, P.B.  et al., Vascular Contributions to Cognitive Impairment and Dementia: A Statement for Healthcare Professionals From the American Heart Association/American Stroke Association, Stroke 42, 2672–2713 (2011).

7. Söderman, M., Andersson, T., Karlsson, B., Wallace, M.C. Edner, G. Management of patients with brain arteriovenous malformations, Eur J Radiol 46, 195–205 (2003).

8. Hendriks, L. et al., Presenile dementia and cerebral haemorrhage linked to a mutation at codon 692 of the beta-amyloid precursor protein gene, Nat. Genet. 1, 218–221 (1992).

9. Kanekiyo, T., Xu, H., Bu, G. ApoE and Ab in Alzheimer’s Disease: Accidental Encounters or Partners? Neuron 81, 740–754 (2014).

10. Verghese, P.B., Castellano, J.M., Holtzman, D.M. Apolipoprotein E in Alzheimer’s disease and other neurological disorders, The Lancet Neurology 10, 241–252 (2011).

11. 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

12.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 in press.

13. Lebrin, F., Srun, S., Raymond, K., Martin, S., van den Brink, S., Freitas, C., Bréant, C., Mathivet, T., Larrivée, B., Thomas, J.L., Arthur, H.M., Westermann, C.J.J.,  Disch, F.,  Mager, J.J.,  Snijder, R.J., Eichmann, A., Mummery, C.L. Thalidomide stimulates vessel maturation and reduces epistaxis in Hereditary Hemorrhagic Telangiectasia patients. Nature Medicine (2010), 16(4):420-8.

14. Orlova, V.V., van der Meer, A.D., Dijke, ten, P., van den Berg, A., and Mummery, C.L. 2013. Three-dimensional co-cultures of human endothelial cells and embryonic stem cell-derived pericytes inside a microfluidic device. Lab Chip. 13:3562-3568.

15. Westein, E., van der Meer, A.D., Kuijpers, M.J., Frimat, J.P., van den Berg, A., Heemskerk, J.W. Atherosclerotic geometries exacerbate pathological thrombus formation poststenosis in a von Willebrand factor-dependent manner. Proc Natl Acad Sci U S A. 2013 110(4):1357-1362.

16. Khairoun, M., de Koning, E.J., van den Berg, B.M., Lievers, E., de Boer, H.C., Schaapherder, A.F., Mallat, M.J., Rotmans, J.I., van der Boog, P.J., van Zonneveld, A.J., de Fijter, J.W., Rabelink, T.J., Reinders, M.E. Microvascular damage in type 1 diabetic patients is reversed in the first year after simultaneous pancreas-kidney transplantation. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons. 2013;13:1272-1281.

17. de Vries, D.K., Khairoun, M., Lindeman, J.H., Bajema, I.M., de Heer, E., Roest, M., van Zonneveld, A.J., van Kooten, C., Rabelink, T.J., Schaapherder, A.F., Reinders, M.E.. Renal ischemia-reperfusion induces release of angiopoietin-2 from human grafts of living and deceased donors. Transplantation. 2013;96:282-289.

18. Khairoun, M., van der Pol, P., de Vries, D.K., Lievers, E., Schlagwein, N., de Boer, H.C., Bajema, I.M., Rotmans, J.I., van Zonneveld, A.J., Rabelink, T.J., van Kooten, C., Reinders, M.E.. Renal ischemia-reperfusion induces a dysbalance of angiopoietins, accompanied by proliferation of pericytes and fibrosis. American journal of physiology. Renal physiology. 2013;305:F901-910.

19. 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.

20. van Solingen, C., Seghers, L., Bijkerk, R., Duijs, J.M., Roeten, M.K., van Oeveren-Rietdijk, A.M., Baelde, H.J., Monge, M., Vos, J.B., de Boer, H.C., Quax, P.H., Rabelink, T.J., van Zonneveld, A.J. Antagomir-mediated silencing of endothelial cell specific microrna-126 impairs ischemia-induced angiogenesis. Journal of cellular and molecular medicine. 2009;13:1577-1585

21. Junker, J.P., Noël, Guryev, V., Peterson, K.A., Shah, G., Huisken, J., McMahon, A.P., Berezikov, E.,  Bakkers, J. and van Oudenaarden, A. Genome-wide RNA tomography in the zebrafish embryo. Cell, in press (2014).

22. Grun, D., Kessert, L. and van Oudenaarden, A. Validation of noise models for single-cell transcriptomics. Nature Methods 11, 637 – 640 (2014).

23. Lekkerkerker, A.N., Aarbiou, J., van Es, T., Janssen, R.A.. Cellular players in lung fibrosis Curr Pharm Des. 2012. 18: 4093-102.

24. Michiels, F., van Es, H., van Rompaey, L., Merchiers, P., Francken, B., Pittois, K., van der Schueren, J., Brys, R., Vandersmissen, J., Beirinckx, F., Herman, S., Dokic, K., Klaassen, H., Narinx, E., Hagers, A., Laenen, W., Piest, I., Pavliska, H., Rombout, Y., Langemeijer, E., Ma, L., Schipper, C., Raeymaeker, M.D., Schweicher, S., Jans, M., van Beeck, K., Tsang, I.R., van de Stolpe, O., Tomme, P. Arrayed adenoviral expression libraries for functional screening. Nat Biotechnol 2002. 20: 1154-1157.

25. Beekers, I., van Rooij, T., Verweij M.D., Versluis M., de Jong N. Acoustic Characterization of a Vessel-on-a-Chip Microfluidic System for Ultrasound-Mediated Drug Delivery.  IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control. Vol. 65, No. 4 April 2018: 570-581 (Cover publication)

26. Halaidych, O.V., Freund, C., van den Hil, F., Salvatori, D.C.F., Riminucci, M., Mummery, C.L., Orlova, V.V. Inflammatory Responses and Barrier Function of Endothelial Cells Derived from Human Induces Pluripotent Stem Cells. Stem Cell Reports vol.10, issue 5, p1642-1656, May 2018

27. Tiemeyer, L.A., Frimat, J-P., Stassen, O.M.J.A., Boutem, C.V.C., Sahlgren, C.M. Spatial patterning of the Notch ligand DII4 controls endothelial sprouting in vitro. Scientific Reports 8 article number 6392, April 2018

28. van Duinen, V., Zhu.D., Ramakers, C., van Zonneveld, A.K., Vulto, P., Hankemeier, T. Perfused 3D angiogenic sprouting in a high-troughput in vitro platform. Angiogenesis 2018, https://doi.org/10.1007/s10456-018-9647-0