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Published on: November 2, 2016
Development of autologous in vitro vascular models for use in preclinical biotherapeutic development
Emma Lund1, Alexander Silvester1, Daniel Thwaites1
1Labcorp, Harrogate, United Kingdom.
Abstract:
Following previous failures to predict drug-induced adverse immune reactions in clinical trials, for example in cases with preclinical species differences or poorly indicative in vitro assays, there has been an emphasis on developing improved preclinical hazard identification tools. Concurrently, there is a regulatory agency-backed responsibility to reduce reliance on preclinical animal models, highlighted by the Food and Drug Administration (FDA) Modernization Act 2.0 and the FDA's 2025 announcement to phase out animal testing for specific compounds. Traditional in vitro cytokine release assays utilize plastic-based formats of antibody presentation to blood cell fractions, and, although biologically simple to run, they do not accurately recapitulate in vivo blood vessel physiology. Including endothelial cells improves physiological relevance by representing the internal vascular wall, enabling cell-cell interactions, compound presentation, and cellular responses from endothelial cells alongside blood cells. Here, endothelial cells outgrown from healthy donors were cocultured with their blood cells to model the immune response to compounds. Building on existing endothelial assays cocultured with blood cell fractions, we established the model using whole blood as an alternative format. We then transferred both formats from 2-dimensional (2D) 96-well plates into a 3D microfluidics system, further mimicking the dynamics and structural microenvironment of a blood vessel. We used these human vasculature models to recapitulate the expected cytokine response to existing compounds and highlight the additional preclinical safety end points that can be investigated by using a 3D vessel, such as vascular leak. This proof-of-concept study demonstrates foundations for a scalable, physiologically relevant method for preclinical testing while reducing reliance on animal models.
Insights
Developing new preclinical models using human cells in 3D microfluidics improves drug safety testing. This approach enhances prediction of immune reactions and reduces reliance on animal testing for drug development.
Area of Science:
- Pharmacology
- Toxicology
- Biomedical Engineering
Background:
- Traditional in vitro assays fail to predict drug-induced immune reactions due to limitations in recapitulating in vivo physiology.
- Regulatory bodies like the FDA are pushing to reduce reliance on animal testing, necessitating novel preclinical hazard identification tools.
- Existing in vitro methods using plastic-based formats do not accurately model blood vessel dynamics and cellular interactions.
Purpose of the Study:
- To develop and validate a physiologically relevant human vasculature model for preclinical drug safety assessment.
- To improve the prediction of drug-induced adverse immune reactions and identify novel safety endpoints.
- To establish a scalable method that reduces the need for preclinical animal models in drug development.
Main Methods:
- Co-culturing endothelial cells with whole blood from healthy donors to model immune responses.
- Adapting 2D cell culture formats to a 3D microfluidics system to mimic blood vessel microenvironment.
- Utilizing human vasculature models to assess cytokine release and vascular leak in response to known compounds.
Main Results:
- The 3D microfluidics system successfully recapitulated expected cytokine responses to existing compounds.
- The model demonstrated the ability to investigate additional preclinical safety endpoints, such as vascular leak.
- The developed human vasculature models showed promise for scalable and physiologically relevant preclinical testing.
Conclusions:
- The 3D human vasculature model offers a more accurate and relevant approach to preclinical drug safety testing compared to traditional methods.
- This innovative model supports regulatory goals by reducing reliance on animal testing while enhancing the prediction of adverse immune reactions.
- Further development of this microfluidic system can significantly advance preclinical safety assessment and drug development pipelines.
