ORCAU Annual Symposium 2023

On Thursday, 23rd of November 2023, ORCAU hosted an annual symposium at the Amsterdam UMC. The programme of the symposium included keynote presentations from experts, a round-table discussion, and poster presentations that highlighted some of the extraordinary work that is being done at the Amsterdam UMC. 

The ORCAU Annual Symposium is intended for researchers who are working with human organoids and organotypic models within the Amsterdam UMC. The symposium serves as an opportunity for researchers to showcase the work that they are doing with human organoids and to network with other scientists working within the field. 

The symposium was opened by Prof. Dr. Hans van Goudoever. The line-up of keynote speakers included Dr. Wendy Dankers, Prof. Eric Reits, Dr. Vivi Heine, and Dr. Harsha Devalla, and panelists of the round-table discussion included Dr. Juan Garcia Vallejo, Dr. Michel van Weeghel, Dr. Peter Henneman, and Prof. Eric Reits. 

Opening by Prof. Dr. Hans van Goudoever
Keynote presentation by Dr. Wendy Dankers
Keynote presentation by Dr. Harsha Devalla
Keynote presentation by Prof. Eric Reits
Prof. Dr. Dasja Pajkrt asking a question during a keynote presentation
Panelists of the round-table discussion

During the event, Debby Weijers, director of Stichting Proefdiervrij (the Dutch Society for the Replacement of Animal Testing ) gave a brief introduction of the Proefdiervrij Venture Challenge; a program that helps scientists set up a venture plan that will push forward animal-free scientific research. She also introduced the winner of the 2023 Venture Challenge, VEO, which consisted of researchers from the Amsterdam UMC.

Debby Weijers introducing the Proefdiervrij Venture Challenge
VEO, Winner of the Proefdiervrij Venture Challenge 202

At the symposium, seven posters were presented by PhD students and technicians working within the Amsterdam UMC. Attendees of the event were encouraged to engage by the posters.  

Nina Johannesson (left) & Eline Freeze (right)
Giulia Moreni (left) & Joep Korsten (right)
Jantine van Voorden
Shicheng Ye
Dr. Katja Wolthers viewing a poster
Prof. Dr. Dasja Pajkrt engaging by a poster
Ellie Wagstaff

We would like to thank Stichting Proefdiervrij, ISOGEN, and STEMCELL Technologies for sponsoring our event. We look forward to hosting the next annual symposium. 

Are you interested in joining ORCAU? Contact organoidcenter@amsterdamumc.nl

Organoid Center Amsterdam UMC (ORCAU) Annual Symposium

We are pleased to announce that on the 23rd of November 2023 at 12 PM, ORCAU will host its annual symposium. This event will take place at Collegezaal 4, Amsterdam UMC location AMC. At this symposium, we have put together an exciting programme highlighting some of the extraordinary organoid work at the Amsterdam UMC. During this event, there will be keynote presentations from experts, a round-table discussion, and an opportunity to present posters. Additionally, the event will kick-off with a lunch and will conclude with a drinks and networking session. 

To see the full programme, click here.

Interested?

If you would like to be a part of the programme, you can submit an abstract before the 30th of September 2023. Please email abstracts to organoidcenter [@] amsterdamumc.nl. For questions, please contact the organization committee using the same email address.

Are you interested in learning more about human organoid technology and how they can be applied in research? Register before the 15th of October 2023 for ORCAU’s upcoming symposium! *Only limited spots available.

About ORCAU

Within Amsterdam UMC, researchers and their team gathered to form an organoid center that enables the use of human organoids and organotypic models for studying human biology and tackling human diseases. The Organoid Center Amsterdam UMC (ORCAU) aims to connect experts in organoid technology, train researchers and disseminate organoid research.

At the moment, ORCAU consists of 14 research groups operating within Amsterdam UMC. These include the Bijlsma Group, CEMM – Molecular Oncology, Complex Ophthalmogenetics, DigeST, Environmental control of the immune system, Immunohematology, Huntington Disease proteostasis group, LEXOR, Lisa van Baarsen Lab, OrganoVIR Labs, Skinlab, RIRE (renal injury and repair), Stem Cell Biology, and Tas Group.

Put Some Guts into It: Intestinal Organoid Models to Study Viral Infection

Abstract

The knowledge about enteric viral infection has vastly increased over the last eight years due to the development of intestinal organoids and enteroids that suppose a step forward from conventional studies using cell lines. Intestinal organoids and enteroids are three-dimensional (3D) models that closely mimic intestinal cellular heterogeneity and organization. The barrier function within these models has been adapted to facilitate viral studies. In this review, several adaptations (such as organoid-derived two-dimensional (2D) monolayers) and original intestinal 3D models are discussed. The specific advantages and applications, as well as improvements of each model are analyzed and an insight into the possible path for the field is given.


Keywords: intestinal organoid; enteroid; intestinal monolayer; Transwell®; Gut-on-a-Chip; Intestine-on-a-Chip; host-virus interactions; enteric virus

Authors

Inés García-Rodríguez, Adithya Sridhar, Dasja Pajkrt, Katja C Wolthers

Link

https://doi.org/10.3390/v12111288

Cerebral Organoids: A Human Model for AAV Capsid Selection and Therapeutic Transgene Efficacy in the Brain

Abstract

The development of gene therapies for central nervous system disorders is challenging because it is difficult to translate preclinical data from current in vitro and in vivo models to the clinic. Therefore, we developed induced pluripotent stem cell (iPSC)-derived cerebral organoids as a model for recombinant adeno-associated virus (rAAV) capsid selection and for testing efficacy of AAV-based gene therapy in a human context. Cerebral organoids are physiological 3D structures that better recapitulate the human brain compared with 2D cell lines. To validate the model, we compared the transduction efficiency and distribution of two commonly used AAV serotypes (rAAV5 and rAAV9). In cerebral organoids, transduction with rAAV5 led to higher levels of vector DNA, transgenic mRNA, and protein expression as compared with rAAV9. The superior transduction of rAAV5 was replicated in iPSC-derived neuronal cells. Furthermore, rAAV5-mediated delivery of a human sequence-specific engineered microRNA to cerebral organoids led to a lower expression of its target ataxin-3. Our studies provide a new tool for selecting and deselecting AAV serotypes, and for demonstrating therapeutic efficacy of transgenes in a human context. Implementing cerebral organoids during gene therapy development could reduce the usage of animal models and improve translation to the clinic.

Authors

Josse A. Depla, Marina Sogorb-Gonzalez, Lance A. Mulder, Vivi M. Heine, Pavlina Konstantinova, Sander J. van Deventer, Katja C. Wolthers, Dasja Pajkrt, Adithya Sridhar, Melvin M. Evers

Link

https://doi.org/10.1016/j.omtm.2020.05.028

Recombination Analysis of Non-Poliovirus Members of the Enterovirus C Species; Restriction of Recombination Events to Members of the Same 3DPol Cluster

Abstract

Enteroviruses (EVs) are highly prevalent viruses worldwide. Recombination is known to occur frequently in EVs belonging to species Enterovirus A, Enterovirus B, and Enterovirus C. Although many recombinant vaccine-derived poliovirus (VDPV) strains have been reported, our knowledge on recombination in non-polio EVs in the species Enterovirus C is limited. Here, we combined a dataset consisting of 11 newly generated full-length Enterovirus C sequences and 180 publicly available sequences to study recombination dynamics in non-polio EVs. To identify recombination patterns, maximum likelihood phylogenetic trees of different genomic regions were constructed, and segregation analyses were performed. Recombination was observed between members of the same 3DPol cluster, but was rarely observed between members of different clusters. We hypothesize that this restriction may have arisen through their different compartmentalization in respiratory and enteric tracts related to differences in cellular tropisms so that the opportunity to recombine may not be available.

Keywords: Enterovirus; recombination.

Authors

Lieke Brouwer, Kimberley S M Benschop, Dung Nguyen, Everlyn Kamau, Dasja Pajkrt, Peter Simmonds, Katja C Wolthers

Link

https://doi.org/10.3390/v12070706

Parechovirus A Pathogenesis and the Enigma of Genotype A-3

Abstract

Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and Listeria monocytogenes were evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research.

Authors

Thomas Roodsant, Marit Navis3, Ikrame Aknouch, Ingrid B. Renes, Ruurd M. van Elburg, Dasja Pajkrt, Katja C. Wolthers, Constance Schultsz, Kees C. H. van der Ark, Adithya Sridhar and Vanesa Muncan

Link

https://doi.org/10.3389/fcimb.2020.00272

Parechovirus A Pathogenesis and the Enigma of Genotype A-3

Abstract

Parechovirus A is a species in the Parechovirus genus within the Picornaviridae family that can cause severe disease in children. Relatively little is known on Parechovirus A epidemiology and pathogenesis. This review aims to explore the Parechovirus A literature and highlight the differences between Parechovirus A genotypes from a pathogenesis standpoint. In particular, the curious case of Parechovirus-A3 and the genotype-specific disease association will be discussed. Finally, a brief outlook on Parechovirus A research is provided.

Keywords: parechovirus, parechovirus A, parechovirus A-3, pathogenesis

Authors

Adithya Sridhar, Eveliina Karelehto, Lieke Brouwer, Dasja Pajkrt and Katja C. Wolthers

Link

https://doi.org/10.3390/v11111062

High frequency and diversity of Parechovirus A in a cohort of Malawian children

Abstract

Parechoviruses (PeVs) are highly prevalent viruses worldwide. Over the last decades, several studies have been published on PeV epidemiology in Europe, Asia and North America, while information on other continents is lacking. The aim of this study was to describe PeV circulation in a cohort of children in Malawi, Africa. A total of 749 stool samples obtained from Malawian children aged 6 to 60 months were tested for the presence of PeV by real-time PCR. We performed typing by phylogenetic and Basic Local Alignment Search Tool (BLAST) analysis. PeV was found in 57% of stool samples. Age was significantly associated with PeV positivity (p = 0.01). Typing by phylogenetic analysis resulted in 15 different types, while BLAST typing resulted in 14 different types and several indeterminate strains. In total, six strains showed inconsistencies in typing between the two methods. One strain, P02-4058, remained untypable by all methods, but appeared to belong to the recently reclassified PeV-A19 genotype. PeV-A1, -A2 and -A3 were the most prevalent types (26.8%, 13.8% and 9.8%, respectively). Both the prevalence and genetic diversity found in our study were remarkably high. Our data provide an important contribution to the scarce data available on PeV epidemiology in Africa.

Authors

Lieke Brouwer, Eveliina Karelehto, Alvin X Han, Xiomara V Thomas, Andrea H L Bruning, Job C J Calis, MichaĂ«l Boele van Hensbroek, Brenda M Westerhuis, Darsha Amarthalingam, Sylvie M Koekkoek, Sjoerd P H Rebers, Kamija S Phiri, Katja C Wolthers, Dasja Pajkrt

Link

https://doi.org/10.1007/s00705-018-04131-7

Polarized Entry of Human Parechoviruses in the Airway Epithelium

Abstract

Human parechoviruses (HPeVs), a poorly studied genus within the Picornaviridae family, are classified into 19 genotypes of which HPeV1 and HPeV3 are the most often detected. HPeV1 VP1 C terminus contains an arginine-glycine-aspartic acid (RGD) motif and has been shown to depend on the host cell surface αV integrins (αV ITGs) and heparan sulfate (HS) for entry. HPeV3 lacks this motif and the receptors remain unknown. HPeVs can be detected in patient nasopharyngeal and stool samples, and infection is presumed to occur after respiratory or gastro-intestinal transmission. HPeV pathogenesis is poorly understood as there are no animal models and previous studies have been conducted in immortalized monolayer cell cultures which do not adequately represent the characteristics of human tissues. To bridge this gap, we determined the polarity of infection, replication kinetics, and cell tropism of HPeV1 and HPeV3 in the well-differentiated human airway epithelial (HAE) model. We found the HAE cultures to be permissive for HPeVs. Both HPeV genotypes infected the HAE preferentially from the basolateral surface while the progeny virus was shed toward the apical side. Confocal microscopy revealed the target cell type to be the p63+ basal cells for both viruses, αV ITG and HS blocking had no effect on the replication of either virus, and transcriptional profiling suggested that HPeV3 infection induced stronger immune activation than HPeV1. Genotype-specific host responses may contribute to the differences in pathogenesis and clinical outcomes associated with HPeV1 and HPeV3.

Authors

Eveliina Karelehto, Cosimo Cristella, Xiao Yu, Adithya Sridhar, Rens Hulsdouw, Karen de Haan, Hetty van Eijk, Sylvie Koekkoek, Dasja Pajkrt, Menno D. de Jong and Katja C. Wolthers. Frontiers in Cellul

Link

https://www.frontiersin.org/articles/10.3389/fcimb.2018.00294/full

A Perspective on Organoids for Virology Research

Abstract

Animal models and cell lines are invaluable for virology research and host–pathogen interaction studies. However, it is increasingly evident that these models are not sufficient to fully understand human viral diseases. With the advent of three-dimensional organotypic cultures, it is now possible to study viral infections in the human context. This perspective explores the potential of these organotypic cultures, also known as organoids, for virology research, antiviral testing, and shaping the virology landscape.

Keywords: human organoids, virology, standardization

Authors

Adithya Sridhar, Salvatore Simmini, Carla M. S. Ribeiro, Caroline Tapparel, Melvin M. Evers, Dasja Pajkrt, and Katja Wolthers

Link

https://doi.org/10.3390/v12111341