12 projects receive grants in 2016
A total of 1 700 000 SEK has been awarded in grants from the Swedish Fund for Research without Animal Experiments in 2016. It was divided between 12 projects. Se recipients and project titles below. More information about the projects will be added shortly.
Patrik Bohman, SLU, receives SEK 50 000 for the project ”Fish, crayfish and mussels as eDNA – test of methodology and usability”.
”Every individual of a species leaves traces of its own DNA in all kinds of environments, for example in lakes and streams. This mixture of DNA is called environmental DNA (eDNA). Today it is possible to detect different species just by taking a water sample and analyze its DNA content, but the methods must be refined and revised. This is a non-destructive methodology that reduces the impact on both the target organisms and their environments. We hope that eDNA sampling will be able to complement more harmful traditional methods, like gillnets and cages. Our vision is therefore to use eDNA as a means to create simple, less harmful, and more cost effective field methodologies to detect fish, crayfish and mussels in natural waters. Even though eDNA will not answer all of our questions (for example age, size and health), we will soon be able to track specific species in lakes and streams merely by sampling water. The support from Forska Utan Djurförsök will help us on our way to achieve our objective.”
Kristina Fant, SP Sveriges Tekniska Forskningsinstitut, receives SEK 200 000 for the project ” Validation of an animal-free method for assessing skin irritating potential of medical devices”.
”All medical devices on the market have to be shown by the manufacturer to be safe for the end-user. The easiest and most common way to do this is to follow the internationally accepted standard series ISO 10993, describing what tests to perform for different types of products and in detail how it should be done. One of the end points that is assessed for more or less all types of medical devices is skin irritation. Despite scientific evidence regarding low predictivity and reproducibility this is still done by the Draize irritation test performed on rabbits, and it is estimated that more than 5000 rabbits are used annually for this test only in Europe. The reason that this method is still so common is the complete lack of other validated methods.
In this project, a novel test method based on three-dimensional human reconstructed skin models is validated in several laboratories in a study coordinated by ISO. Validation is the link between method development and regulatory acceptance and use. It consists of a thorough evaluation of the method’s reproducibility, accuracy, repeatability and predictivity, performed through multi-site evaluation of several different medical device materials of known irritancy. This is the last step required before the test method can be formally accepted and included in the standard series, so that the animal testing can be stopped without hampering the development of novel medical devices.”
Anna Forsby, Stockholm University, receives SEK 100 000 for the project ”Mapping genomic biomarkers from neuroblast to terminal differentation in a human cell modell”.
Information about the project will be provided shortly.
Martin Hallbeck, Linköping University, receives SEK 250 000 for the project ”Development of a platform to test compounds that may be candidates for stopping Alzheimer’s disease”.
”The project utilise the stem cells developed in the Herland project described below. By developing animal free research models that are better than animal experiments there is a great hope that many animal experiments can be replaced.
The goal is to develop a platform to test substances that could become drugs against Alzheimer’s and other neurodegenerative diseases. Despite the best efforts we still lack crucial knowledge about Alzheimer’s disease, the most common cause for dementia, and we lack a cure. It’s known that the disease spreads from one part of the brain to the next. However, the mechanisms for this is not known although there is a growing understanding that small, toxic aggregates of misfolded proteins are important. The cognitive function of the patients deteriorates with the spread of the disease. Eventually becoming dependent on others and many dies in advance.
Because the living human brain is difficult to access, good models are needed to understand and combat brain diseases. The research group works with human cells in model systems that they have developed to study how Alzheimer’s and other neurodegenerative diseases can spread between neurons. They have developed the method further by using human induced pluripotent stem cells (iPSC). These cells have been created in another project that receives funding from Forska Utan Djurförsök (see Anna Herlands project) by reprograming donated mature skin cells. If they can show that this model can be used by the pharmaceutical industry and for research about dementia with better results than by using animal experiments, many animal experiments could be replaced. In addition, the understanding about these devastating diseases could accelerate. The preliminary results are promising!”
Anna Herland, Karolinska Institutet, receives SEK 200 000 for the project ”Stem cell derived cholinergic neurons as in vitro models to reduce animal experiments within neurodegeneration and analgesia”
”The aim of our research is to create better model systems of human brain tissue. Such models are essential to increase knowledge about function of cell types found in the brain as well as to facilitate development of new drugs. We are focused on research within analgesia, drugs that relieve pain, and neurodegeneration. These disease conditions are prevalent, for example it is estimated that one in five Europeans suffer from chronic pain and that about 5.4 million Americans are diagnosed with Alzheimer’s disease.
The available models systems for neuronal tissue used today are animals, especially rodents, and human, cancer derived, cell lines. Both systems are strikingly bad at predicting both effects and side effects of drugs. The guiding hypothesis of this project is that differentiation of stem cells in to neuronal cells in combination with three-dimensional cell culture systems can provide highly relevant models of the human brain.
We are using human induced pluripotent cells (iPS cells) originating from human skin biopsies. These cells can be differentiated to virtually any specialized cell in the human body; we have developed efficient methods to make them into neurons. By culturing the differentiated neurons in three-dimensional, gel like systems we can recapitulate disease specific phenomena that can not be seen in conventional cell culture systems. We have furthermore derived iPS cells from skin biopsies from Alzheimer’s patients and when we generate neurons from these cells they show signature of the disease after a time period of a few weeks. The Alzheimer specific neurons we generate are easily used multi-parallell screening applications and could be valuble for drug development. Ultimately our systems could decrease the demand to use animals or animal derived cells both in academic and industrial settings.”
Maria Karlgren, Uppsala University, receives SEK 150 000 for the project ”A humanized cell model for accurate assessment of molecular transport across the blood-brain barrier”.
”Developing drugs for treatment of CNS diseases is more difficult as compared to development of drugs for other therapeutic areas. This is due to the blood-brain barrier (BBB) which protects the brain from entry of drugs and other xenobiotics. In drug development in vitro and in vivo animal models are used for prediction of BBB permeability. However, these models don’t translate very well to humans. The reason for this is that the drug transport proteins, which are present in the BBB and are important for the barrier function, differs between species. Hence, today the role of human drug transporters in the BBB is often overlooked. This project aims to solve the problem by developing predictive models that take the human transporters expressed at the BBB into account. With these novel models we will be able to both estimate drug delivery to the human brain and to identify the mechanisms involved. At present we have e.g. developed a cell model completely lacking transporter background. This in an important step in the BBB model development, but it has also a wide applicability within other research fields. We are convinced that this model as well as other refined models developed in this project will expand the technical platform and provide more predictive and high-throughput BBB models than currently available and thereby reduce the number of animal based experiments in drug development.”
Agnete Kirkeby, University of Lund, receives SEK 100 000 for the project ”A 3D in vitro model of the human fetal brain”.
Although the complexity and size of the human brain is vastly different from that of animal models, studies on brain development are today almost exclusively limited to rodents and other smaller animals. The animal models are used because they are easily accessible, and because it is not possible to do genetic and dynamic studies on brain development in humans. This in turn also means that we have poor knowledge about the aspects of brain development which makes the human brain so complex. In this project, we will develop a 3D in vitro model of the early developing human brain through the use of human pluripotent stem cells and advanced microfluidic culturing techniques. The project is the result of an innovative collaboration between stem cell scientists and bioengineers at Lund University. By exposing stem cells to reconstructed growth factor gradients in the dish, we are building a microenvironment in which the stem cells will spontaneously form anatomical structures resembling the early stages of the human fetal brain. We envision that the model in the future can be used as an assay for screening neuromodulating and toxic effects of environmental chemicals and medical compounds relevant to human fetal development.
Pär Matsson, Uppsala University, receives SEK 150 000 for the project ”Models for predicting intracellular drug exposure”.
”For a pharmaceutical drug to give the desired effect it must reach the organ and the cells that need treatment. In most cases, the pharmacological effect results from that the drug molecule binds to a specific protein inside the target cell. To predict if a new drug will give the desired effect it is thus necessary to determine the amount of drug that is distributed into the different cells of the body.
We will combine our recently developed experimental method, which allows measurements of intracellular drug exposure in cultured human cells, with advanced imaging techniques. Together with novel modeling techniques, this will allow measurement and prediction of drug exposure in subcellular compartments – knowledge that is of key importance in understanding the pharmacological and toxicological response to drugs.
Drug-like molecules that do not reach their subcellular targets in sufficient amounts, or which result in dangerously high concentrations, can thereby be filtered out in the early stages of the drug development process, avoiding unnecessary experiments in laboratory animals and patients.”
Stina Oredsson, University of Lund, receives SEK 100 000 for the project ”Novel Cell Culturing Methods to Reduce Animal Use in Cancer Reserach ”.
”Cancer is one of the leading causes of morbidity and mortality with an expected rise over the next decade due to life style factors and an aging population. Research concerning the cause of cancer and to find new efficient treatment strategies, without side effects are highly prioritized. A tumour in the body grows in a 3D environment interacting with normal cells which is different from the flat 2D environment that cancer cell lines are grown in. To mimic the 3D environment of the body to hopefully achieve more relevant information regarding new treatment strategies, we will develop a 3D cell culturing system for co-cultivation of cancer cells and normal cells. This co-culturing system will be derived using a minimum amount of animal-derived products. Instead of using collagen, we will use electrospun fibres and instead of using the extracellular matrix (ECM) component matrigel (derived from a mouse sarcoma), we will use recombinant ECM components including hydrogel molecules such as hyaluronic acid. We have already successfully co-cultured cancer cells and fibroblasts in such a system in medium without fetal bovine serum, another animal-derived product that raises serious ethical concern. We will also add other normal cells to the culture such as lymphocytes and incubating the 3D system in transwells will allow co-culturing with e.g. hepatocytes to include investigation of the role of drug metabolism. Besides replacing animal-derived products, our goal is to suggest a 3D test system including normal cell toxicity before new potential anti-cancer compounds can be tested in animals.”
Lena Palmberg, Karolinska Institutet, receives SEK 100 000 for the project ”A three-dimensional model based on human cells from the airways to replace animals for controlled nanoparticles aerosol exposures”.
“Chronic bronchitis is associated with impaired health status, acute exacerbations, increased risk of development of chronic airflow obstruction (COPD) and a poor prognosis when occurring in patients with COPD. It is therefore of utmost importance to explore the patho-physiological mechanisms behind the condition. By using advanced 3D-models with human primary bronchial epithelial cells of both a normal mucosa and a chronic bronchitis-like mucosa and combine those with immune effector cells, these mechanisms can be evaluated in more details and different treatment strategies can be evaluated. 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, University of Skövde, receives SEK 100 000 kr for the project ”Stem cell-derived human cardiomyocytes as an alternative to animal models in toxicity assessment”.
Our research aims towards the development of an in vitro model based on cardiomyocytes derived from human pluripotent stem cells. This model is expected to be very valuable in order to reduce the number of animals used for drug development and risk assessment of chemicals.
The specific focus for this project is to evaluate the usefulness of human stem cell-derived cardiomyocytes, as a relevant alternative to animal studies, to detect and in detail study drug-induced cardiotoxicity. As a toxic model compound doxorubicin will be studied.
Doxorubicin is an effective chemotherapeutic agent that unfortunately is associated with severe cardiac side effects. Despite intense research, the exact mechanisms responsible for doxorubicin-induced cardiotoxicity are not known. The model systems available today are typically based on the use of animal models, which represent limited value for the study of human toxicity. The use of primary human cardiac tissue is challenging due to a very low availability of donated material, which often also is variable and associated with a history of disease and drug treatment.
In our experimental system, the stem cell derived human cardiomyocytes are exposed to doxorubicin for up to 48h and then we monitor the cells during a wash out period of up to 14 days. The global protein expression will be measured using quantitative proteomics (LC-MS/MS and TMT-labeling). Bioinformatic algorithms will be applied to the data in order to identify differentially expressed proteins in the doxorubicin exposed cells. The proteomics data will also be 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.
We expect that this study will lend further support to the use of human stem cell-derived cardiomyocytes as a relevant model system to investigate cardiotoxicity in vitro.
Lena Svensson, Lund University, receives SEK 250 000 for the project ”Replacing animal models by microfluidic vascular models to study cell migration”.
”We plan to use “lab on a chip”-devices, to create a more physiologically relevant models of the vascular system, thereby creating a better replacement for today’s studies using animals. We will, in short, culture endothelial cells in microchannels, covering an extra cellular matrix mimic, creating a model interior blood vessel wall. This system will enable us to perform microscopy based mechanistic studies of cell migration, on and through the vessel wall, and other cell functions, something which animal models do not readily lend themselves to.
Cell migration through the blood vessels is key function in the immune system whereby leukocytes can spread to a site of infection to deal with foreign threats. Dysfunctional leukocyte migration is of importance in several diseases such as autoimmune diseases. Cellular transmigration is also a key aspect in cancer progression: cancer metastasis is the process during which cancer cells leave their original tumour, move through the surrounding tissue and enter the vascular system. The cancer cells travel through the vascular system and eventually cross the vessel walls (extravasation) and establish new tumours, with detrimental consequences to the patients, causing more than 80% of cancer related deaths. The mechanisms involved in this process are poorly understood and current knowledge relies heavily on animal experiments, either as an end point assay or as highly invasive intravital microscopy.
The development of a more relevant and advanced human physiological model will increase the number of researchers who will choose to go from animal models to human in vitro models. This will not only lead to reduced unnecessary use of animals in the academic setting and pharmacology industry research community but also lead to refined experimental control, giving the user control over several parameters not possible in an in vivo setting.”
Senast uppdaterad: 25 maj 2016
