Grants awarded to 15 projects for 2022
A total of 3,6 million SEK has been awarded in grants from the Swedish Fund for Research without Animal Experiments for 2022, divided between 15 projects.
Read the researchers own descriptions of their projects below.
Tomas Bergström, Gothenburg University, 200 000 SEK Project: Development of antiviral drugs against respiratory viruses without animal experiments
Respiratory viruses such as Respiratory Syncytial virus (RSV) and Coronaviruses (CoV) such as COVID-19 are imminent threats to global health. No effective antiviral treatment exist to these both virus groups.
We have discovered novel antiviral targets for RSV in form of EGF receptors (unpublished), and interactions between the non-structural protein 6 (nsp6) and Sigma-like receptors for CoV (Lundin A et al., PLoS Pathogens 2014;10(5):e1004166 and unpublished).
We intend to develop these lead compunds to antiviral drugs. To this end, we have established air-liquid interface respiratory cell cultures of human origin for antiviral testing. This modell builds a multi-layered cellular structure resembling the human airway epithelium, with retained ciliary function. These cells are grown with human growth factors and other defined chemical cell medium compounds, instead of calf serum and other animal products.
The object of the current project is to test the antiviral efficacy and cellular toxicity of our lead compunds against RSV and CoV in these ALI cultures. If these tests are successful, we intend to pursue clinical phase 1 trials in humans without animal experiments. The basis for this ambition is that no animal models are fully relevant for these two human viruses, which instead display their pathogenicity in the ALI model, which thus is better suited for evaluation. At our lab, we have BSL3 facilities in which experiments on COVID-19 are performed. experiments on RSV and other CoV are performed at BSL2 level. If successful, clinical application of the findings of the project could be fast, given the current pandemic situation.
Malin Celander, Gothenburg University, 250 000 SEK
Project: FishTox22 – Developing fish cell-line based tox tests to replace fish toxicity tests
The thousands of chemicals we use every day all require testing for their safety to the environment. In Europe this is carried out under the REACH legislation and through approval procedures by national and EU authorities such as EMA for medicines and EFSA for chemicals in the food chain.
Assessment of safety of chemicals to the environment usually involves toxicity testing on fish and in the EU about 180 000 fish per annum are used in chemical toxicity testing.
There is societal demand to reduce and eventually replace fish toxicity testing of chemicals, which will require the development of in vitro and in silico alternatives. The US NIH Tox21 project has led the way by demonstrating that cell-based toxicity testing is more efficient and can be as precise in testing of chemicals as traditional animal studies. However, methods to assess the safety of chemical to the environment are still largely relying on animal studies and most frequently LC50 assays.
In this international collaborative project, we aim to develop new in vitro tox-test-models for environmental risk assessment using end-points in cell-lines from three different fish species (zebrafish, rainbow trout & guppy), to replace current fish toxicity tests.
There are currently no protocols to culture fish cells entirely without the use of serum in the media. Therefore, we also aim to develop new protocols with serum-free media for tox-tests in these cell-lines.
The project will provide new insight in systems toxicology, and we test the hypothesis that our cell-based assays can predict toxic potential of chemicals and correctly identify toxicity pathways. This project has scientific significance as new knowledge will be generated on effects of 10 chemicals with different Modes-of-Action (MoA) on several Adverse Outcome Pathways (AOPs).
In addition to replacing fish toxicity testing, the information generated can be used to train in silico models to predict toxicity of chemicals in the environment to fish.
Anna Erlandsson, Uppsala University, 300 000 SEK
Project: Investigating the role of astrocytes in Alzheimer’s disease, using human cell culture models
Alzheimer’s disease (AD) is the leading cause of dementia. Yet, no treatments that limit neurodegeneration or slow down the disease progression are available. Hence, innovative therapeutic approaches are clearly required.
The key neuropathological hallmarks of AD are extracellular plaques, mainly consisting of aggregated Aβ, intracellular neurofibrillary tangles, composed of hyperphosphorylated tau and chronic inflammation.
Decades of research have focused on neuronal abnormalities during AD progression, while glial cells, including astrocytes, have been given much less attention.
The aim of this project is to clarify the involvement of astrocytes in AD and to investigate their therapeutic potential by using advanced human cell culture models. Our previous results demonstrate that astrocytes engulf large amounts of aggregated amyloid-beta and tau, but then store, rather than degrade the ingested material. This incomplete degradation results in severe cellular stress and the release of extracellular vesicles containing neurotoxic content, which could be of relevance for AD progression.
To further investigate the impact of astrocytes in amyloid-beta and tau pathology, we will use cell culture models that mimic the central disease processes in the human AD brain. For this purpose, we will develop different co-cultures, close-cultures and 3D organoids of human iPSC-derived astrocytes, neurons and other relevant cell types.
The cell models will be used to:
• Examine the role of astrocytes AD progression by spreading of pathological protein aggregates.
• Elucidate if antigen presenting astrocytes are driving the chronic inflammation in AD.
• Identify ways to reduce astrocyte-mediated neurotoxicity and disease spreading.
Being the most numerous glial cell type in the brain, astrocytes constitute a compelling treatment target. Our results will contribute with important data and hopefully pave the way for novel therapeutic strategies to halt AD progression.
Anna Falk, Karolinska Institute, 250 000 SEK
Project: Organoids as hosts and semi-in-vivo models of the human brain to reduce and replace animals
This is an extension of a previously approved project with the main objective of this 3R program is to develop human organoid systems that will act as hosts and “in-vivo-like” brains for transplanting cells to replace the use of animals.
There are two main reasons to develop brain organoid systems that mimics the human brain, one being that animal brains have been shown to be different to humans and thus sub-optimal for studies of brain disorders, second now with emerging stem cell technologies we have the chance and the responsibility to develop human “in-vivo-like” brain models for research and thereby replace animals.
We in my group are stem cell biologists with long experience in reprogramming and stem cell models of the brain. Here we would like to use our knowledge to create organoid models of the human brain that will be used as a substitute for animal brains for transplantation of cells for cancer studies, disease modelling of neuronal maturation and for cell replacement therapy studies. The three different types of transplantation schemes we would like to investigat
e and develop are the following (listed in the order of number of animals that can be replacement), organoids as hosts: a) For transplantation of tumour cells for studies of brain cancer b) For transplantation of patient derived neural cells to investigate differentiation, migration, integration, maturation into a healthy “brain-like” environment c) For studies on neural cells replacement in for example organoid models of Parkinson’s disease
We have previously standardised stem cell culture and neuronal differentiation using defined, serum free and xenofree reagents, which creates robust stem cells showing close to none variability. Here we will develop the brain organoid system by replacing Matrigel with the defined and animal free 3D material BioSilk. The produced organoids will be investigated for their ability to host cell transplantation and thereby replace animal brains.
Marianne Jansson, Lund University, 300 000 SEK
Project: Animal-free lymphoid infection; clues to why HIV-2 is less aggressive and blunts HIV-1?
The HIV-1 epidemic is still spreading. Thus, the need for a vaccine or cure is urgent. To identify virus-host mechanisms that could be targeted for cure strategies a fraction of HIV-1 infected individuals with spontaneous virus control, elite controllers, have been studied.
However, only 0.15% of HIV-1 infected individuals are elite controllers. In contrast, elite controllers among those infected with HIV-2, a benign form of HIV, has been estimated to 37%. Therefore, HIV-2 infection may represent a better model for finding cure strategies. In an epidemic study we showed that HIV-2 also delays HIV-1 AIDS development with 90% in HIV-1/HIV-2 dually infected. Furthermore, the blunting effect that HIV-2 has on HIV-1 progression was observed to be an early HIV-1 infection event.
We now want to identify mechanisms behind HIV-1 and HIV-2 differences, and how HIV-2 suppresses HIV-1. Since HIV primarily replicates in lymphoid tissue, infection experiments using blood-derived lymphoid cells do not provide a clinically relevant model. Humanized mouse and non-human primate models are therefore increasingly used to study early infection events taking place in lymphoid tissue.
To avoid use of animal models, we here aim to establish a novel model where infection of lymphoid tissue, from human tonsils, is used to dissect early events of HIV-1, HIV-2 and dual infections. The tonsil tissue will be infected with HIV-1 and HIV-2, individually and dually, and gene specific chimeric viruses will be constructed for deciphering virus-specific determinants of replicative capacity and host interactions.
The replication of the viruses, as well as the expression of genes and proteins in different cell populations, infected and bystander, will be analyzed using advanced bioinformatics including network analysis and modeling. By this we hope to contribute to the development of animal free infection models and strategies to prevent AIDS development in HIV-1 infection.
Per Malmberg, Chalmers Technical University, 200 000 SEK
Project: Chemical imaging for skin permeation studies – an analytical approach to replace animal experiments
We have recently shown that our animal-free skin permeation analysis approach can be used to study uptake of active pharmaceutical compounds (APIs) as well as to study metal uptake in skin for sensitization studies. In this project we want to validate our method and compare to existing an OECD approved artificial skin models.
We also wish to study if the model can be used to evaluate environmental effects such as the penetration of persistent organic pollutants, airborne particles and microplastics through the skin. The aim for the project is that the model can replace or significantly reduce existing in-vivo models and improve on already established in-vitro models. This by offering a more sensitive and flexible analytical approach that can replace and/or complement existing methods in the OECD guidelines for skin adsorption (no 427, no 428 and no 429) and measure metals and APIs simultaneously in the skin while being able to also trace endogenous effects in cells.
Ivan Nalvarte, Karolinska Institute, 250 000 SEK
Project: A new model to study Alzheimer’s disease in human mini-brains
Experimental research on Alzheimer’s disease (AD) is extensively based on animal models due to the lack of relevant and reliable in vitro methods that recapitulate the disease progression. However, human disease relevance of animal models has been questioned, warranting the development of alternative models for human AD.
The hallmarks of AD include cortical and hippocampal deposition of amyloid plaques, neurofibrillary tangles and activation of local immune response that all contribute to extensive neuronal death and cognitive decline.
The proposed continuation-project aims at developing an immune competent in vitro AD model that recapitulates this pathology and that can be used in high-throughput.
Cerebral brain organoids from induced human pluripotent stem cells harboring either driving AD mutations and controls or chemically induced amyloidogenesis will be used. These will be grown in a tailored simple high-throughput microgravity bioreactors to allow long term culturing. To address the impact of immune response on AD pathogenesis, we will establish co-cultures of cerebral organoids with isogenic microglia.
This model will be validated against human AD brain samples available in brain banks. In this way, we aim at replacing molecular studies using animal AD models by offering a human relevant in vitro model, thereby greatly reducing the number of mice included in AD research today.
Mats Nilsson, Stockholms Universitet, 300 000 SEK
Project: The Design of a Glioblastoma Organoid-on-a-Chip for Drug Testing
Glioblastoma multiforme (GBM), in addition to being the most common type of malignant primary brain tumor, is one of the tumors that has seen the least improvement in treatment options in the last half-century. The existing tools we have (mouse models, cell culture models) for testing efficacy are insufficient and inefficient.
New tools are needed. Specifically, a tool that tests a drugs ability to pass the BBB and reach the tumor, that recapitulates the GBM tumoral environment and complexity, while allowing efficacy to be assessed in the context of biomarkers. Toward that end, the project proposal is to develop an in vitro glioblastoma organoid on-a-chip that can be integrated into the drug discovery systems of tomorrow, and used to assess preclinical effectiveness of drugs on glioblastomas.
Elin Nyman, Linköping University, 250 000 SEK
Project: Computer models within inflammation that reduce the need of animal experiments
The process of inflammation is important to understand since it is a key component in many common diseases: cancer, cardiovascular diseases, and infectious diseases such as covid-19 to mention a few. Even though substantial efforts have been made, there is a lack of a mechanistic understanding of both shared hallmarks of inflammatory processes, and mechanistic differences of importance for common diseases.
To study mechanisms, computer models has shown potential since they allow for quantitative hypothesis testing of proposed mechanisms. Developed models can be simulated to predict the outcome for new experiments and therefore used in knowledge-based experimental design. Such simulations can be extended to the scale of a whole population, where individual differences are included and therefore can be used to evaluate the effects of new drugs for different groups of individuals. Computer models can therefore bridge between detailed in vitro data and clinical observations – without the use of animal experiments.
In a collaboration between researchers at Örebro University, Linköping University, and AstraZeneca, the aim is to set up such a knowledge driven workflow to study the process of inflammation. We aim to extract and build functional general models of the process of inflammation, i.e. models that would be shared among both different organs and tissues and potentially also shared between different diseases. These general models can then be specialized in different ways to answer different clinically relevant questions about inflammation.
In summary, this project will use computer models together with human-derived in vitro and clinical data from the inflammatory process. The models will serve as a basis for a knowledge driven workflow, where mechanistic knowledge can be connected to clinical outcome, and where the use of animal experiments become superfluous.
Penny Nymark, Karolinska Institute, 200 000 SEK
Project: Toward risk assessment without animals: Development of Adverse Outcome Pathways for lung cancer
The risk for chemicals and nanomaterials to cause cancer is complex to assess and currently based mainly on animal experimentation, even though non-animal alternative methods exist. Some of these cell-based methods and assays are accepted by regulatory instances for risk assessment, however, are mainly applied as weight of evidence information in addition to currently required animal data. Furthermore, novel high-throughput systems, capable of handling increasing numbers of samples in short time frames, require validation towards regulatory acceptance.
The new Adverse Outcome Pathway (AOP) concept is expected to simplify and improve the integration of alternative methods towards becoming gold standard regulatory accepted methods for risk assessment. AOPs describe toxic mechanisms causally connected to human organ injury and disease. They provide a basis for development of mechanism-focused alternative methods predictive of the risk for toxicity.
This project will continue to develop and refine an AOP for lung cancer, which has been developed with funding from Forska utan Djurförsök. The project will apply toxicological and data management expertise with the aim to integrate big data derived from a wide number of diverse nanomaterials, as well as to support efficient reuse of the project results.
Novel data handling and integrative approaches will be applied in line with the recently established FAIR principles, which guide scientific approaches in order to make results Findable, Accessible, Interoperable and Reusable. Implementation of FAIR principles effectively supports efficient reuse and application of results in practical settings.
The overall aim of the project is to develop, facilitate and support the integration of AOP-aligned animal-free methods for cancer risk assessment of inhalable nanomaterials.
Lena Palmberg, Karolinska Institute, 200 000 SEK
Project: Development of lung models with multiple cell types for treatment strategies of chronic lung disease
Air pollutions caused 9 million premature deaths in 2015. Chronic obstructive pulmonary disease (COPD) can be caused by air pollution and is the third leading cause of death worldwide. By using multi-cellular lung mucosa models with human primary bronchial epithelial cells, combined with immune effector cells (macrophages), air pollution related health hazards will be assessed. In the research team pre-clinical and clinical experts as well as scientist from the pharma industry warrant a comprehensive assessment of scientific methodology and data interpretation. The goal includes evaluation of different treatment strategies.
The studies explore the interaction of exposure, therapeutic effects, innate immunity, protease/antiprotease balance and oxidative stress as well as the interactions of various cell types. This knowledge can be utilized to develop preventive measures and treatment strategies for patients with COPD and chronic bronchitis. Validation of the models enables us to develop a systematic in vitro-testing strategy during drug development in order to reduce the requirement for animal inhalation studies.
Nicolas Pillon, Karolinska Institute, 230 000 SEK
Project: In silico comparative profiling of skeletal muscle immunometabolic responses
The rapid advancement in technology has allowed wide-scale use of ‘omics’ in human and many model organisms, but the amount of biological data generated surpasses the ability of researchers to comprehensively process it. We here propose to add “reuse” as a fourth “R” to the 3R principles of animal experimentation.
The overarching aim of this project is to promote the reutilization of data with the objective to reduce the need for preliminary exploratory analyses in animal models. We will use in silico analyses to decipher subtypes of immunometabolic responses in skeletal muscle to define novel interventional strategies that extinguish or promote selective components of inflammation to improve metabolism.
This project will develop innovative bioinformatic approaches taking advantage of publicly available data to identify novel therapeutic targets and generate accessible omics databases of human and mouse. The cross-species comparison of the response of skeletal muscle will provide a complete map of the mouse and human responses to exercise and metabolic diseases and therefore help increase translatability of exercise and metabolic research.
Jane Synnergren, Skövde University, 200 000 SEK
Project: Reduce the need for animal models by advanced 3D in vitro cardiac hypertrophy disease model
Animal models have been required to increase our understanding of the pathogenesis and underlying causative mechanisms associated with cardiac hypertrophy (CHT). Rodent models have been used for decades as disease models for CHT, but do not fully recapitulate the human CHT pathophysiology.
In recent years, human pluripotent stem cells-derived cardiomyocytes (hPSC-CMs) have emerged as a powerful tool to overcome the limitations of using animal models to study CHT. hPSC-CMs can be produced at high scale and purity for disease research and drug development. However, a major hurdle for their application is the immature state of the hPSC-CMs. Thus, more knowledge about the developmental drivers of hPSC-CM maturation is needed in order to implement strategies to enhance the maturation of hPSC-CMs.
Recent data show that advanced 3D culture methods using bioprinting, organoids, and multi-cell type culturing improve the maturation and functional properties of hPSC-CMs.
This project will extend our work on 2D CHT disease models and use bioprinting technology to develop a more advanced 3D CHT disease model based on CMs, cardiac fibroblasts and endothelial cells of human origin. To induce CHT characteristics the model will be exposed to endothelin-1.
The 3D disease model will provide novel means to explore the dynamic changes and remodeling that characterize CHT progression, and identify causative molecular mechanisms associated with cardiomyocyte maturation. Development of advanced CHT disease models would greatly benefit from the generation of fully mature hPSC-CM since it will allow a more accurate recapitulation of the CHT disease phenotype in an in vitro setting, replacing current animal models.
The project will also collect large-scale data from in vitro models and explore how digital models can be used to simulate key mechanistic for CHT progression, and investigate how concept of digital twins, digital organs and digital cells can reduce the need of animal models.
Brun Ulfhake, Karolinska Institute, 220 000 SEK
Project: 3R Ranker: a search wizard for replacement of animal testing
By the one-year grant (N2020-0009) we will be able to deliver a search engine for Replacement alternatives in late 2021. As described below the project is on-track and set-out milestones will be delivered by the end-of-2021 or early 2022 (quarter 1). The open access functional search engine to be delivered will be hosted at one of the participating sites (Hanze University of Applied Science, TenWise, NL): https://kmine.tenwiseapps.nl/
The project was presented at WC 11 in Maastricht 2021.
As described in the original application as a second step, the search engine should be more rigorously validated and further refined, with web design and learning tools. The current application for a continued funding 2022 covers these aspects and will complete the project mission. To facilitate the implementation of the 3Rs, we will develop software for rapid and effective searches for identifying Replacement alternatives, by applying artificial intelligence methodologies.
Our specific aim is to create 3R data framework with the following properties:
1. Powered by a machine learning model that automatically retrieves 3R related papers from the scientific literature stored in the MEDLINE database. The focus will be on Replacement.
2. Automatically updated with new daily releases of the MEDLINE database.
3. Attached to a cloud-based graphical user interface (webserver) that allows querying with > 200.000 biological keywords such as genes and diseases.
4. Linked to a database with curated, validated Replacement research papers.
The methodology will be developed, validated and refined based on data from the field of skin irritation and neuroscience. The software will be made freely available for use and educational purposes. Our initiative also includes a training tool for users of this resource.
Lena Öhman, Gothenburg University, 250 000 SEK
Project: Human intestinal organoids as a tissue based model system for studying cellular mechanisms of intestines
Many diseases of the gastrointestinal tract are multifactorial and there is not one single experimental animal model that resembles all aspects of human conditions, irrespective of disease.
During recent years, the importance of the interaction between the host and the local intestinal microenvironment, thus the microbiota and metabolites they produce, for disease generation has become evident. T
he aim of this project proposal is therefore to develop a tissue-based model system making use of our in house developed intestinal organoids for determining epithelial events giving rise to intestinal diseases, alongside assessing cellular mechanisms of therapeutic drugs.
Our unique model is based on organoids from healthy donors, which are stimulated with fecal supernatants containing luminal factors, from patients and healthy subjects, giving rise organoids with different characteristics, representing disease-like and healthy-like “mini-guts in a dish”. Thus, our hypothesis, supported by preliminary data, is that organoids cultured in the presence of fecal supernatants reproduce the intestinal conditions of the donors who provided the fecal material. The effects of fecal supernatants on organoids are determined by microscopy, RNA and protein profiles.
This organoid system will allow us to determine events of the epithelium perpetuating disease as well as mechanisms of therapeutic drugs. The outcome of this project proposal include a tissue based model system, adequately reproducing both the healthy and diseased state of the intestinal tissue, providing a useful tool for studying and comparing the early epithelial events in different intestinal diseases.
The model system may also be used for investigating cellular mechanisms of therapeutic drugs. The tissue based model system will in part replace experimental animal models exploring epithelial events of human intestinal diseases such as irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) and colon cancer.
Senast uppdaterad: 5 juli 2023
