Experts

Monique Janssens (personal account): Why we need the Transition towards Animal-free Innovations
Expert interviews
HelpathonsPolicy

Monique Janssens (personal account): Why we need the Transition towards Animal-free Innovations

Why is there a Transition towards Animal-free Innovations, while we have the 3Rs, including Replacement? Well, there is a difference. Animal experiments should no longer be the golden standard of reference. We should not ask: Is this animal-free method good enough to replace animal experiments? But: What is the research question, and how do I get the best answer, preferably without animals? I know that many researchers are doing this already. But we can do even more! It’s also about involving the full chain of parties, including patients, financers, legislators and companies. That is why the transition movement works with interdisciplinary networks and Helpathons. The transition helps to innovate, to accelerate and to implement. At the same time, there is no need to throw the 3Rs overboard. Actually, we owe applying them to the lab animals of today. But by innovating we can develop even more new practices in research and education that bring about better results for science in less time and often with less costs. Without using animals.
02:376 months ago
Daniela Salvatori: TPI Utrecht
Expert interviews
HealthEducation

Daniela Salvatori: TPI Utrecht

Prof. dr. Daniela Salvatori, chair of TPI Utrecht, presents the aims of the local TPI group and invites all who want to share their ideas or questions on the transition towards animal-free innovations to get in touch via uu.nl/tpi.
02:2314 months ago
Elly Hol (UMC Utrecht): possibilities for neuroscience
Expert interviews
HealthInnovation

Elly Hol (UMC Utrecht): possibilities for neuroscience

Prof. dr. Elly Hol (neuroscientist) talks about the opportunities for conducting animal-free research in Utrecht. She explains why it is necessary to use animal models next to cell-based models, for example for her Alzheimer research. More info at https://www.umcutrecht.nl/en/research/center/brain-center , http://translationalneuroscience.nl , http://www.ellyhollab.eu .
00:5915 months ago
Wim de Leeuw: Aim and activities of TPI Utrecht
Expert interviews
EducationInnovation

Wim de Leeuw: Aim and activities of TPI Utrecht

TPI Utrecht facilitates the Utrecht infrastructure to stimulate the transition to animal-free innovation. There is a helpdesk, a group of ambassadors, and a 3Rs Stimulus Fund. A digital marketplace for exchange of tissues is being developed, and as well a hybrid centre for biomedical translation, where there will increasingly be place for animal-free techniques.
01:5617 months ago

Innovation

Respiratory toxicity using in vitro methods
Innovation examples

Respiratory toxicity using in vitro methods

The airways form a barrier for inhaled compounds, however, such compounds may cause local effects in the airways or may lead to lung diseases, such as fibrosis or COPD. Cell models of the respiratory tract, cultured at the air-liquid-interface (ALI) are a relevant model to assess the effects of inhaled compounds on the airways. Such models allow human relevant exposure, which is via the air, and assessment of effects on the epithelial cell layer. At RIVM we use air-liquid-interface cultured cell models and expose these to airborne compounds to assess the effects of agents such as nanomaterials, air pollutants or compounds from cigarette smoke. By using a mechanism-based approach to assess the effects of these compounds we invest in animal-free alternatives that better predict adverse effects in humans.
02:242 months ago
FirstbaseBIO - human brain organoids for studying neurological diseases
Innovation examples
HealthInnovationIn vitro

FirstbaseBIO - human brain organoids for studying neurological diseases

Human neurological diseases are still poorly understood, amongst others because animals are used as a model for the human brain. A way to overcome this problem is to mimic human brain functioning in a dish with organoids. FirstbaseBIO is developing off-the-shelf brain organoids on which neurological diseases can be studied. This 3D platform will be formed by reprogrammed human cells from easily accessible sources, for example urine, skin, or mucosa. The proof of-concept brain organoids will be those from patients who are suffering from adrenoleukodystrophy (ALD), a rare, incurable brain disease that occurs primarily in young boys and is often fatal. With the brain organoid platform, possible medicinal treatments for ALD can be effectively optimised. FirstbaseBIO was nominated for the Venture Challenge 2021 for their development of human brain organoids to study neurological diseases.
03:337 months ago
GUTS BV - small intestine-on-a-chip and advanced computational analysis for compound and protein screening
Innovation examples
HealthToxicologyIn vitro

GUTS BV - small intestine-on-a-chip and advanced computational analysis for compound and protein screening

GUTS BV is a contract research organization offering its 3-dimensional state-of-the-art small intestinal in vitro model in combination with custom computational analysis approaches. The small intestinal model was developed during Dr. Paul Jochems PhD research at Utrecht University in the group of Prof. Roos Masereeuw. In comparison to the current gold standard (Transwell model), they show improvement in cell differentiation (all major specialized cell types present), physiological structure (3D tube- and villi-like structures) and a functional epithelial barrier. After acquiring experimental data from this model computational analysis approaches are used to score and compare measured compounds for all tested biological parameters at once. The combined effort of improved in vitro modelling and data analysis is believed to result in an enhanced preclinical predictability. GUTS BV was nominated for the Venture Challenge 2021 for their development of an intestinal model combined with advanced computational analysis for protein and chemical compound screening. Research papers: https://www.sciencedirect.com/science/article/pii/S0887233318307811 https://www.mdpi.com/2072-6643/12/9/2782/htm https://www.nature.com/articles/s41538-020-00082-z LinkedIn: https://www.linkedin.com/company/71016128/
02:117 months ago
Avatar Zoo - teaching animal anatomy using virtual reality
Innovation examples
EducationInnovation

Avatar Zoo - teaching animal anatomy using virtual reality

Animals are essential to train the next generation of scientists understand diseases and develop treatments for humans as well as animals. Therefore, animals are used for educational purposes. Technologies such as Virtual Reality and Augmented Reality can be employed to reduce the number of animals in the future. Prof. Dr. Daniela Salvatori is working on the development of 'Avatar Zoo' together with UMCU and IT. Live animals are replaced by holographic 3D in this flexible platform. With these holograms one is able to study the anatomical, physiological and pathological systems and processes of all kinds of animals. Avatar Zoo won the Venture Challenge 2021 for the development of virtual reality models that can be used for anatomy classes and practical training.
02:407 months ago

Meetings

Helpathon #4 - can you help Frank?
Meeting videos
HelpathonsHealth

Helpathon #4 - can you help Frank?

Can you help Frank with integrating an immune system into a macaque lung organoid to address local immunity to tuberculosis with his vaccination strategy? Join Helpathon #4, look at www.tpihelpathon.nl/coming-up ! Frank Verreck does research on tuberculosis at the Biomedical Primate Research Center (BPRC). Tuberculosis is the most deadly infectious disease worldwide! For the past hundred years, BCG (Bacillus Calmette Guérin) vaccinations take place through the skin. Research shows that macaques can be better protected from this infection by vaccination through their lungs. Frank really wants to further study the potential of this alternative vaccination strategy. He wants to understand how this BCG vaccination works in macaques lungs.
01:2117 months ago
Helpathon #4 - can you help Raissa?
Meeting videos
HelpathonsIn vitro

Helpathon #4 - can you help Raissa?

Can you help Raissa find a more complex organoid-like brain and immune model based on rhesus microglia to study aging in relation to neuroinflammation and neurodegenerative diseases? Join Helpathon #4, look at www.tpihelpathon.nl/coming-up ! Raissa Timmerman is a PhD student at the alternative unit at the Biomedical Primate Research Center. A better understanding of aging of the brain is key to studying neuroinflammation and neurodegenerative diseases. We believe there is a potential for breakthrough in using our existing live macaque data obtained from past aging experiments to develop more complex in vitro rhesus brain-like models and then to correlate all this data with data from human in vitro models and human live data.
01:3217 months ago
Helpathon #4 - can you help Anne-Marie?
Meeting videos
HelpathonsIn vitro

Helpathon #4 - can you help Anne-Marie?

Can you help Anne-Marie develop a more organ-like Rhesus 3D liver model in which she can study the dormancy and the waking up of malaria parasites? Join Helpathon #4, look at www.tpihelpathon.nl/coming-up ! Anne- Marie Zeeman is a researcher at the Biomedical Primate Research Center (BPRC). Anne-Marie studies recurrent malaria ( P. vivax). She successfully developed a single cell layer in vitro model to study compounds affecting dormant and active malaria parasites in the liver of Rhesus monkeys. We believe that the cross correlation between in vitro Rhesus and in vitro human models will provide the missing link required to improve the drug development process and aid transition. A more refined Rhesus in vitro model can reduce the number of monkeys currently used for testing drugs. The data from in vivo monkeys combined with new in vitro models could help validate and develop reliable human in vitro models making testing on monkeys unnecessary detours.
01:3817 months ago
Sign in for Helpathon #3: Saskia van Mil
Meeting videos
HelpathonsHealth

Sign in for Helpathon #3: Saskia van Mil

Saskia van Mil calls for a Helpathon! She invites you to help her develop a human model for studying liver metabolism? You can sign in for this Helpathon here: tpihelpathon.nl. Online, 18th - 19th of June 2020.
01:1620 months ago

Conferences

Scientific solutions for the gap in translational medicine: skin model platform with melanoma (3D melanoma)
Conferences abstracts

Scientific solutions for the gap in translational medicine: skin model platform with melanoma (3D melanoma)

The developing process of a new drug, from first testing to regulatory approval and ultimately to market is a long, costly, and risky path. Noteworthy is the fact that almost 95% of the drugs that go into human trials fail. According to the National Institutes of Health (NIH), 80 to 90% of drug research projects fail before they ever get tested in humans. The value of preclinical research, mainly conducted in animal model experiments for predicting the effectiveness of therapies and treatment strategies in human trials, has remained controversial. Only 6% of the animal studies are successfully translated into the human response. Breaking down failure rates by therapeutic area, oncology disorders account for 30% of all failures. The absence of human-relevant models with receptors, proteins, and drug interactions in the in situ microenvironment leaves a gap in the scientific discovery process of new therapies. In this context, the present work presents the development of a sophisticated in vitro skin model platform focus on boosting melanoma treatment. The results showed a physiological microenvironment of human skin with epidermal differentiation and development of stratified layers (basement membrane, stratum spinosum, stratum granulosum, and stratum corneum). Furthermore, it was observed the pathophysiological microenvironment of the melanoma with invasion or migration through the basement membrane into the dermis and no epidermal differentiation. Vemurafenib treatment, the gold standard which targets BRAF mutations, showed a decrease in proliferation and invasion of melanoma tumors, with an increase in epidermis keratinization. Melanoma incidence continues to increase year-on-year and is currently responsible for >80% of skin cancer deaths. It is the most common cutaneous form and is known to have the highest mutational load of all cancers. Nowadays, patients with advanced melanoma BRAFV600E mutation can benefit from monotherapies or targeted therapies. Although the initial response rate is effective, disease progression and tumor chemoresistance rapidly occur in the majority of patients. Therefore, the treatment of melanoma remains a challenge, and despite the advances, there is still an urgent need to identify new therapeutic strategies. 3D Model Melanoma is considered one important tool for studying the evolution of the pathology, as well as evaluating the effectiveness of new therapeutic approaches.
03:1915 days ago
Optimizing CAR-T-cell therapy using 3D tumor models and real-time cell imaging
Conferences abstracts

Optimizing CAR-T-cell therapy using 3D tumor models and real-time cell imaging

Chimeric antigen receptor (CAR) T-cell therapy accounts for one of the most promising therapeutic advances in cancer immunotherapy. In this form of adoptive cell transfer, T-cells of a patient are engineered to express so-called ‘CARs’, in which the antigen-recognition capacity of antibodies is combined with T-cell activating domains. So far, CAR-T-cell therapy obtained its most impressive results in hematological malignancies resulting in the approval of five CAR-T cell products by the FDA for hematologic indications. However, CAR-T-cell therapy has not mirrored its success in solid tumors. The poor efficacy of CAR-T-cell therapy in solid tumors has, in part, been attributed to the lack of understanding in how CAR-T-cells function in a solid tumor microenvironment. Classical validation methods rely on the use of specificity and functionality assays in 2D models against adherent target cells or target cells in suspension. Yet, by using these models, observations made in vitro may differ greatly to an in vivo situation where tumors are engrafted in 3D structures. We developed a more relevant and translational 3D tumor model using eGFP+ target cells. This allows us to couple 3D tumor cell killing by CAR-T-cells to live-cell imaging, providing an efficient quantification of target cell death. As proof- of-concept, we used a 3D model of eGFP+ glioblastoma cells and CAR-T-cells targeting a pan-cancer antigen. This 3D glioblastoma model allowed us to show that classical scFv-based CAR-T-cell therapy of glioblastoma cells can be improved by nanoCAR-T-cells. Furthermore, combining nanoCAR-T-cell therapy with a genetic approach of nanobody-based anti-PD-L1 immune checkpoint blockade further increased the cytotoxicity of the nanoCAR-T-cell therapy.
03:2515 days ago
Biotransformation of two proteratogenic anti-epileptics in the zebrafish (Danio rerio) embryo
Conferences abstracts

Biotransformation of two proteratogenic anti-epileptics in the zebrafish (Danio rerio) embryo

The zebrafish (Danio rerio) embryo has gained interest as an alternative model for developmental toxicity testing, which still mainly relies on in vivo mammalian models (e.g., rat, rabbit). However, cytochrome P450 (CYP)-mediated drug metabolism, which is critical for the bioactivation of several proteratogens, is still under debate for this model. Therefore, we investigated the potential capacity of zebrafish embryos/larvae to bioactivate two known mammalian proteratogens, carbamazepine (CBZ) and phenytoin (PHE) into their mammalian active metabolites, carbamazepine-10,11-epoxide (E-CBZ) and 5-(4-hydroxyphenyl)-5-phenylhydantoin (HPPH), respectively. Zebrafish embryos were exposed to three concentrations (31.25, 85, and 250 μM) of CBZ and PHE from 51⁄4 to 120 hours post fertilization (hpf) at 28.5°C under a 14/10 hour light/dark cycle. For species comparison, also adult zebrafish, rat, rabbit and human liver microsomes (200 μg/ml) were exposed to 100 μM of CBZ or PHE for 240 minutes at 28.5°C. Potential formation of the mammalian metabolites was assessed in the embryo medium (48, 96, and 120 hpf); pooled (n=20) whole embryos/larvae extracts (24 and 120 hpf); and in the microsomal reaction mixtures (at 5 and 240 minutes) by targeted investigation using a UPLC–Triple Quadrupole MS system with lamotrigine (0.39 μM) as internal standard. Our study showed that zebrafish embryos metabolize CBZ to E-CBZ, but only at the end of organogenesis (from 96 hpf onwards), and no biotransformation of PHE to HPPH occurred. In contrast, our in vitro drug metabolism assay showed that adult zebrafish metabolize both compounds into their active mammalian metabolites. However, significant differences in metabolic rate were observed among the investigated species. These results highlight the importance of including the zebrafish in the in vitro drug metabolism testing battery for accurate species selection in toxicity studies.
02:5715 days ago
Lung tumor spheroids for onco-immunological research
Conferences abstracts

Lung tumor spheroids for onco-immunological research

Lung cancer thrives in a complex multicellular tumor microenvironment that impacts tumor growth, metastasis, response, and resistance to therapy. While orthotopic murine lung cancer models can partly recapitulate this complexity, they do not resonate with high-throughput immunotherapeutic drug screening assays. To address the current need for relevant and easy-to-use lung tumor models, we established a protocol for fully histo-compatible murine and human lung tumor spheroids, generated by co-culturing lung fibroblasts with tumor cells in ultra-low adherence 96-well plates. Moreover, we describe their application potential to study tumor-stroma organization, T-cell motility, and infiltration as well as distinct macrophage subsets’ behavior using confocal microscopy. Finally, we report on a 3D target specific T-cell killing assay that allows spatio-temporal assessment using live cell imaging and flow cytometry. This lung tumor spheroid platform can serve as a blueprint for other solid cancer types to comply with the need for straightforward onco-immunology assays.
03:0916 days ago