Drug Discovery & Development 2020

Whether you are performing killing assays, characterizing the tumor microenvironment, or investigating immunogenicity, primary cells are essential for the discovery of new therapeutics. Conclusive and relevant assay results depend on reliable sample preparation, including Ficoll-free cell separation and efficient tumor dissociation. We have developed cell isolation solutions from various starting materials to ensure the best fit for every drug discovery program - join us to learn about the best methods and processes for your in vitro assays.

Learning Objectives:

1. Understand the fundamentals of MACS technology and magnetic cell separation

2. Discuss how the automated isolation of primary cells leads to more efficient and reproducible  in vitro assays

3. Use of primary cells in drug discovery assays

Digital spatial profiling (DSP) on the NanoString GeoMx platform enables high plex molecular profiling of clinical tissue samples. This new profiling strategy enables deep insights into the heterogenous nature of gene and protein expression as it is driven by the distribution and colocalization of cells within a tissue. Recently, DSP was featured in 2 back to back Nature publications elucidating the important role of B cells in mediated immune response to immunotherapy in melanoma. These studies, from the laboratories of Dr. Jennifer Wargo at the MD Anderson Cancer Center and Dr. Goran Jonsson at Lund University, leverage DSP as well as other traditional and next generation profiling strategies to deeply characterize the molecular expression patterns and spatial architecture of the key immune populations within tertiary lymphoid structures in the tumor. Dr. Sarah Warren will speak about both of these publications, as well as how the work was enabled by prior DSP profiling of melanoma samples from Dr. Wargo’s lab. This webinar will present an overview of the GeoMx platform as well as demonstrate its application to current clinically impactful questions in immuno-oncology.

Learning Objectives:
1. Understand the biology around B cells and tertiary lymphoid structures in cancer
2. Understand how spatial high plex profiling was used to characterize immune responses within melanoma

3D liver microtissues (also known as 3D spheroids) have the potential to positively impact drug development workflows, particularly when applied to safety assessment and drug discovery processes. These 3D in vitro liver models, which can be engineered to mimic healthy and diseased biology in human or model organisms (e.g., monkey, dog, and rat),  have been proven to be powerful tools for fulfilling the demands of modern toxicology programs, both in terms of providing translational safety insights and pushing the bench closer to the patient bedside.
 
In this lecture, you will learn:
 
1. Practical liver toxicology applications for 3D human liver microtissues, from safety testing to complex disease modeling, using non-alcoholic steatohepatitis (NASH) as an example.  
2. Considerations for adapting existing protocols to leverage 3D liver microtissues and upgrade your tox toolbox for investigating specific scientific questions related to drug induced liver injury (DILI) and drug efficacy testing of new drug candidates, such as promising therapies currently under development for NASH —all with a high degree of flexibility.
3. Guidelines for adopting 3D human liver microtissues for use in high-throughput applications, which require uniform spheroid quality, functional robustness, assay scalability, and reproducibility of results.
4. How 3D cell technology is evolving into an investigative tool that will meet the challenges of the next generation of therapeutic modalities.

The use of primary cells in vitro is compromised by the limited quantity of cells that can be isolated from one donor, a lack of or very restricted proliferation capacity (e.g. hepatocytes) and/or the change from a quiescent to an activated state (e.g. hepatic stellate cells). The use of immortalized cell lines, on the other hand, is compromised by their transformed, cancerous phenotype. Using the upcyte® technology, primary cells are driven into proliferation using a viral gene transfer system, thus allowing controlled and reversible bypass of cell cycle control mechanisms without inducing immortalization, uncontrolled cell growth or changing the typical phenotype. In this talk liver-specific cells such as upcyte hepatocytes and liver sinusoidal endothelial cells will be presented.

Learning Objectives:

1. Identifying different cell types of the liver and their roles

2. advantages and disadvantages of genetic modification of primary cells

The OrganoPlate is a microtiterplate based Organ-on-a-Chip platform for high throughput drug safety and efficacy screening. It contains up to 96 cell culture  chambers that allow co-culture of 3D tissues under continuous perfusion. Highly physiologically relevant tissue and disease models are constructed by (co-)culturing human cells in 3D under continuous perfusion whilst maintaining ease of use and compatibility with standard equipment. This presentation will highlight several applications of the OrganoPlate including models of vasculature, gut, kidney, liver, brain and tumor physiology including iPS and organoid based tissues. The models are interrogated using a variety of assays including (Confocal) Immunocytochemistry, calcium imaging, barrier integrity assessment, qPCR and TEER measurements.

Learning Objectives:

1. Introduce the audience to the fields of organ-on-a-chip and micro physiological systems for preclinical research

2. Explain current tissue culture modalities and (high throughput) assays available in organs on chips

The liver plays a critical role in the metabolism and clearance of more than 70% of marketed drugs. Furthermore, toxicity to the liver is a major reason for preclinical and clinical drug failures. Animals are not always predictive of human liver-drug interactions due to significant species-specific differences in drug metabolism and toxicity pathways. Thus, in vitro models of the human liver are now routinely utilized in pharmaceutical practice to complement animal testing; such models enable faster and cheaper drug testing and hold the promise to prevent harm to living patients. Isolated primary human liver cells are the gold standard for fabricating human liver models, but these cells rapidly lose their liver functions, including drug metabolism capacities, in conventional 2-dimensional monocultures. In contrast, semiconductor-driven microfabrication tools and co-culture with non-parenchymal cells have been used to precisely modulate the microenvironment around hepatocytes towards enhancing and stabilizing phenotypic functions for 4-6 weeks. Such engineered liver co-cultures have significantly improved the sensitivity for the detection of drug clearance, drug metabolite identification, drug-transporter interactions, and drug-induced liver toxicity. This presentation will showcase the development and validation of engineered human liver co-cultures for the above-mentioned applications; it will then discuss emerging trends in further improving the physiological relevance of the platform using liver non-parenchymal cells and determining the role of major liver diseases (e.g., non-alcoholic fatty liver disease) in changing drug outcomes.

Learning objectives:
1. The key design features necessary for building robust in vitro human liver models and
2. The various applications within the drug development pipeline where such models can be effectively utilized.

Microphysiological systems (MPS), also known as organ-on-chips, are small scale in vitro cell cultures which mimic facets of tissue or organ level function. MPS frequently utilise primary human cells, often cultured in 3D, to obtain highly functional, physiologically relevant models. MPS can be utilized alone, but can also be connected through fluidic circuits to create advanced multi-MPS that can model the interactions between organ systems, allowing greater analysis of molecular pathways and disease mechanisms.

Most current in vitro liver models are simple mono-cultures of hepatocytes or use hepatic cell lines and do not have the complexity to analyse the interactions between different tissue types and determine how these interactions drive specific pathologies. We have developed a novel system for the in vitro culture of hepatocytes in a perfused three-dimensional format, with/without a separate co-culture allowing interactions to be studied with a second MPS. The model of the human liver, contains multiple primary cell types, which can be cultured together for several weeks and maintain their phenotype and metabolic activity. The MPS platform provides a tool to study in greater detail the key interactions between the gut, the lungs, the liver and the immune system that drive pathologies including steatohepatitis, hepatocellular carcinoma and viral hepatitis.

Learning Objectives:

1. Introduction to the field of microphysiological systems

2. Demonstrate use of MPS for efficacy, tox and ADME applications

“The poor translatability of early-stage preclinical models is a major setback in oncology drug development. Immortalized cell lines, that are extensively used in drug screens, undergo genetic drift over time due to adapting to in vitro growth, thus showing limited relevance to the human disease. Interpatient tumor heterogeneity is also a major hurdle with standard in vitro models failing to recapitulate this variability. Patient-derived xenografts are the gold standard preclinical models for their proven translatability to the human disease but can be costly and time consuming to generate and maintain. The Hubrecht Organoid Technology (HUB) has pioneered the development of tumor organoids, a revolutionary patient-derived 3D in vitro system that recapitulates the genomic, morphological, and pathophysiological characteristics of their parental tumor. HUB organoids provide highly clinically relevant 3D in vitro models for oncology drug development. To overcome the limitation imposed by sourcing patient material for organoid preclinical development PDX can be used as a source of human tumor tissue leading to a patient-relevant matched in vitro/in vivo platform for oncology drug screening.”

Learning bullets:
1. Find out why HUB organoids are considered best in class systems
2. Understand how organoids are revolutionizing oncology drug development
3. Learn about how patient-derived xenografts (PDX) can be leveraged to develop the largest organoid library for oncology drug discovery

With an increasing push to improve safety, efficacy, and efficiency throughout the drug development pipeline, researchers are evermore looking to improve the predictive capacity of their in vitro models. By employing 3D cell culture models (i.e. spheroids, organoids), which better recapitulate the native tissue environment in comparison to traditional 2D cultures, disease states can be mimicked more effectively, and thus therapeutics can be evaluated in a more meaningful context. Yet the use of these more complex 3D cell culture models stipulates a more sophisticated method of analysis in order to extract the full potential of these models. In this webinar, Erin Edwards of Visikol will discuss 1) how 3D cell culture models can be designed and applied to evaluate new therapeutics, and 2) how tissue clearing and 3D high content confocal imaging can be used to extract the most meaningful conclusions from studies involving 3D cell culture models. Examples will be given from toxicity evaluation (neural, cardiac, liver) and traditional and immuno-oncology.

Learning Objectives:
1. Understand the differences, pros, and cons of each 2D and 3D cell culture models
2. Learn about specific applications in drug discovery where 3D cell culture models can afford advantages over traditional 2D cell culture models.