2017

13 projects receive grants in 2016

A total of 2 million SEK has been awarded in grants from the Swedish Fund for Research without Animal Experiments in 2017. It was divided between 13 projects. Se recipients and project titles below. More information about the projects will be added shortly.

 

Martin Andersson, SP Technical Research Institute of Sweden
Animal-free method for prediction of eye-irritation 
In an internal project SP / RISE, we have surprisingly found that it seems possible to predict whether a liquid is an Eye-irritant or not via the liquids so-called Hansen parameters. Each liquid has three such parameters describing the solubility properties of the liquid, and therefore each fluid can be assigned a place in ”Hansen-space” where the three parameters are x, y and z coordinates. Also polymers have specific Hansen parameters and quids / polymers that have similar parameters are miscible / dissolve in each other. By viewing the cornea as a polymer, we have been able to develop a first animal free-test method that looks promising – based both on in-silico predictions as well as on a test where a specific polymer thread is dipped into the liquid to be asessed, followed by measurement of the elongation of the thread. In case of a Eye irritant that irritates via impregnation into/swelling of the cornea we have seen thread-elongations of up to 50%.
In this project we will further develop this method so that we can ensure that it works and correlates with the real eye-irritation data and we will also try to develop it further so that it can predict the clouding of the cornea and even eye damage caused by acidic / basic or oxidizing / reducing substances

Kunal Bhattacharya, IMM, Karolinska institute: Development of a lung-on-chip model for testing of nanotoxicity
Engineered nanomaterials (ENMs) are being produced for an increasing number of smart technology applications. Therefore, it is important to perform safety assessments of these ENMs to mitigate possible human exposures and health-risks. Due to their small size the ENMs have larger and more reactive surface as compared to their bulk counterparts, and may have yet to be discovered mechanisms of inducing toxicity in humans. In 2013, the Swedish Government issued a report on the safe handling of ENMs and recommended to set aside funding for research and development of testing and risk assessment methodology. Currently, simple 2-D in vitro cell culture models dominate the field of nanotoxicology research, which do not mimic the complex 3-D multicellular microenvironment of tissues and organs. Mechanistic studies performed in small animals such as rats and mice also do not represent a real-life human exposure due to phenotypic, genotypic and physiological differences. Finally, the local microbiota cooperates with the host environment and this is missed in conventional in vitro 2D cell cultures. Therefore, in the proposed research, we aim to develop a microfluidics-based complex tissue system representing human lungs (lung-on-chip devices) using human cell lines grown in serum-free conditions. These models will be used for testing the toxicity of silver and silica nanoparticles. Results generated using the lung-on-chip model will be used to identify the mechanistic toxicity of these ENMs and will be correlated to previously published research work using 2D in vitro models, as well as published in vivo results. The study outcome will provide evidence regarding the usefulness of the lung-on-chip model as a replacement for the existing 2D in vitro and in vivo study models used in the field of nanotoxicology and help in enhancing knowledge on the toxicity of ENMs on human health and development of chronic/acute diseases.

Gunnar Cedersund, Linköping university:

Pernilla Eliasson, Linköping university: How do tendons heal? – and what happens when they don´t heal?
The ethology behind tendon pathology is multifactorial, however different drugs might influence this. Tendon injuries are common and require a long rehabilitation time. Mechanical loading improves healing, but it is unclear how. I have introduced a model for 3D cell culture studies of human primary tendon fibroblasts, in Linköping. This 3D model makes it possible to study effects on mechanical strength of the tissue after for example drug treatment or changes in loading situations. This is in contrast to conventional cell culturing. Studies on mechanical strength is an important tool in orthopedic research and this has previously only been done in animal models.

Moreover, this in vitro model also allows me to study mechanotransduction. Mechanotransduction is the way a cell converts mechanical stimulus into a biochemical response. I will use this model to study the effect of loading without micro-damage associated inflammation (which is present in animal models) to investigate the mechanotransduction component during loading. I have previously observed that tendon fibroblasts from some patients lack the ability to form tendon tissue, in vitro. I will study the differences between cells that can form tendon tissue with those that can´t. This might help to identify factors that are vital for tendon formation.
I have also started a project where I study some aspects on why tendon injuries appear. We will perform epidemiological methods to study associations between statin use and tendon injuries in different part of the body. I will also use my in vitro model to study the mechanisms why tendons are affected by statins and high cholesterol levels.
The goal with my studies is to understand some factors which might contribute to the appearance of different tendon injuries, and to gain information to increase the understanding on what happens after a tendon injury, so that treatment and rehabilitation protocols can improve.

Martin Hallbeck, Linköping university: Utveckling av en plattform för att testa substanser som kan vara kandidater för att bromsa Alzheimers sjukdom

Maria Karlgren, Inst f pharmacy, Uppsala university: Human cell based models for accurate predictions of brain drug uptake
Poor predictions of CNS drug exposure is a major problem in CNS drug development and is primarily the result of relying on animal-based models although major species differences are well-known for the blood-brain barrier (BBB) and the expression of drug transporters. Here, we will solve this problem by developing predictive models taking the drug transporters expressed in human BBB into account and thereby accurately quantify drug delivery to human brain.
This will be done by development of mechanistic transporter models. Thereafter in vitro kinetics from mechanistic transporter models and human BBB in vitro models will be combined with quantitative proteomics and physiological parameters to predict the human in vivo situation. The project will be performed at Uppsala University in close collaboration with UDOPP (part of DDDp/SciLife Lab) giving a unique opportunity for validation in real-life drug development projects.
With our model drug candidates with predictable and favorable human BBB properties will be identified and selected early in drug discovery/development. As a result an initial reduction of animal PK experiments with 50-60%, with further reduction potential, is estimated. Thereby providing the refined tools needed to substantially reduce and, in a longer perspective, replace animal BBB studies throughout the drug discovery/development process.

Pekka Kohonen, IMM, Karolinska institute: Deepened mechanistic validation of toxicity pathway functionality in a patented analysis tool for toxicity prediction

Johan Lundqvist, The Swedish University of Agricultural Sciences, Uppsala: Establishment of animal-free cell models for studies of aquatic toxicity
This research program aims to develop a system for animal-free testing to assess aquatic toxicity, based on the principle of toxicity pathways. Today, European regulations require that a large number of chemicals should undergo assessment of aquatic toxicity, which often include animal experiments. Assessment of the aquatic toxicity in toxicity pathway-based cell culture models instead of in animal experiments, will allow analysis of a large number of chemicals to a moderate cost and will significantly reduce the number of animals used for aquatic toxicity assessment of chemicals. The European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM) has recently proposed development of cell cultured based assays as a key strategy to reduce and replace the use of fish in toxicity testing.
The aims of this project are

• To establish a panel of 10-15 robust toxicity pathway-based assays that are measuring toxic effects of high relevance for fish toxicity assessment

• To use the established toxicity pathway-based assays to study the toxicity of approx. 25 chemicals that has already underwent classical animal-testing for aquatic toxicity, and to compare the results from the in vitro-assays with the results from the already performed animal experiments

• Based on the results from this project, we will suggest a panel of toxicity pathway-based assays that can be used as an initial screening step for aquatic toxicity and provide information to prioritize which substances that needs to undergo further toxicological investigations

Pär Matsson, Inst f pharmacy, Uppsala university
Methods for predicting intracellular drug exposure
Current methods for predicting the effect of new drugs do not adequately address that drug concentrations can differ substantially between the circulating blood and the cells where the drug target is expressed. We have developed a rapid method for measuring unbound drug concentrations in the interior of cultured cells. The specific aims of this project is to 1) further develop the method and validate it for prediction of unbound drug exposure in different cell and tissue types, 2) to use the method to explore how transport proteins affect intracellular drug concentrations, 3) to study if the method can be used to improve predictions of modulation of intracellular drug targets such as protein kinases, and 4) to develop computational models that can predict intracellular drug exposure from the chemical structure alone.

Results so far: An experimental method for intracellular concentration measurements was developed and applied to the human cell line HEK293, and to primary human hepatocytes. The method was shown to give comparable results to previously described methods that use tissues from laboratory animals, and we also show that cell binding measurements in the HEK293 cell line can be used to predict binding in several different human tissues, including the liver and the brain. Studies of the link between drug transport proteins and intracellular concentrations (Aim 2) and of how unbound drug concentrations affect intracellular drug effects are ongoing and expected to be finalized during the coming funding period. Using the data obtained so far, computational models were developed that accurately predict cellular binding for new drug compounds, without the need for experiments.

Stina Oredsson, Lund university: Novel 3D Cell Culturing Methods in Cancer Research -For Better Prediction, Efficacy, and Less Animal

Lena Palmberg, IMM, Karolinska institute: New lung models with primary cells and exposure to components in air pollutants
The primary aim is to develop and evaluate unique and relevant normal and chronic bronchitis-like airway wall models with primary human bronchial cells from healthy subjects and smokers with and without COPD. The sophisticated airway wall models will be co-cultured with innate effector cells (neutrophils and alveolar macrophages) from different patient groups and mimic the in vivo-situation better than animal models where species differences always is an issue. These models with multiple cell types enable us to study cell-to cell interactions and cross-talk between the cells. By using our advanced exposure systems we can expose cell cultures to airborne particles, only consuming minimal amounts of test substance if the access is limited. Our chosen test substances of different ambient air pollutants like biodiesel particles, nanoparticles and aldehydes contribute to the development of asthma, chronic bronchitis, COPD and cardiovascular diseases and trigger symptoms in subjects suffering from those diseases. Development and validation of these models enable us to develop an in vitro testing strategy in order to reduce the requirement for animal inhalation studies. The applicant team has long experience of studies of in vivo-effects induced by exposure in an exposure chambers of the above mentioned agents in healthy subjects and subjects with respiratory diseases. Therefore the clinical relevance of the models will be well evaluated.

Peter Sartipy, Systems Biology Research Center, School of Bioscience, University of Skövde: Stem cell derived human cardiomyocytes as in vitro model for toxicity testing
Our research aims towards the development of an in vitro model based on cardiomyocytes derived from human pluripotent stem cells. Such a model is expected to be very valuable in the future, in order to reduce the number of test animals used for drug development.
The specific focus for this project is to evaluate human stem cell-derived cardiomyocytes, as a relevant alternative to animal studies, to detect and in detail study drug-induced cardiotoxicity using doxorubicin. Doxorubicin is an effective chemotherapeutic agent that is associated with severe cardiac side effects. The model systems available today are mostly based on the use of animal models, with a limited value for the study of human toxicity. The use of primary human cardiac tissue is problematic due to a very low availability of donated material, which often also is associated with a history of disease and drug treatment.
The cardiomyocytes are studied during treatment with doxorubicin for up to 48 hours. The global protein expression is measured using quantitative proteomics. Bioinformatic algorithms are applied to the data in order to identify differentially expression proteins in the doxorubicin exposed cells. The proteomics data is also integrated with already available transcriptomics data (global mRNA and microRNA expression data) in order to identify affected signaling pathways and mechanisms linked to the toxicity.
The results show a clear effect of doxorubicin on the cells protein expression. A number of vital functions are affected within the cells, such as proteins linked to the cardiomyocytes contractility, apoptotic signaling, energi metabolism, and synthesis and modulation of proteins. Interesting differential expression patterns that show a linkage between the proteome, transcriptome, and the regulatory microRNA network, were identified. Findings that help to increase the understanding of the mechanisms behind and suggest putative biomarkers for anthracycline-induced cardiotoxicity.

Lena Svensson, Lund university: Mikrofluidiska blodkärl istället för djurmodeller för att studera cellmigration

 

 

Senast uppdaterad: 27 mars 2017