Review by the Editor
Researchers gather for interactive forum
Metrion Biosciences and Axol Bioscience joined forces on Wednesday 23rd May to co-host the 2018 “Interactive Stem Cell Forum” in Cambridge (UK). A meeting featuring a morning of informative and thought-provoking talks from leading academic and industry-based researchers working in the stem cell field, followed by an afternoon of laboratory demonstrations in Metrion’s Granta Park headquarters.
The meeting enabled researchers from across Cambridge and the surrounding area to attend an event focused solely upon recent developments in the field of stem cell research spanning neuroscience and cardiac topics. It was also an opportunity for attendees to network with each other and also see demonstrations utilising Axol’s Induced Pluripotent Stem Cells (iPSCs) in Metrion’s laboratories; where the Metrion team also showcased manual patch electrophysiology, QPatch 48 automated electrophysiology and microelectrode array (MEA) assay platforms.
Review of available techniques applied to iPSC research
The morning session, chaired by Metrion Biosciences’ CSO Marc Rogers, started with a presentation by Matthew Daniels, a Consultant Cardiologist based at the University of Oxford. Matthew described some limitations of technologies that have previously been applied to iPSC research. For example chemical dyes such as BAPTA-AM and Fura-2 have significant cellular toxicity, evidenced by such dyes negatively effecting the contractility properties of iPSC-derived cardiomyocytes.
Matthew is a strong advocate for the adoption of alternative fluorescent/luminescent tools and optogenetic-based stimulation for iPSC cardiomyocyte research, he provided a thorough review of the available techniques and described some of his research into non-invasive phenotyping and drug screening. Use of such technology has enabled the monitoring of cells for periods of up to 90 days in his laboratory. He also presented preliminary results using microarrays of single cells on a single assay plate that may have potential for scale up and use in drug discovery.
The objectives of the CiPA initiative
The co-hosting companies also gave data-led presentations, with Metrion’s Sarah Williams discussing the establishment of Axol’s cardiomyocytes as a model system using manual patch clamp electrophysiology in Metrion Biosciences laboratories. Sarah reviewed the objectives of the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative, a multi-agency strategic enterprise with the objective of improving cardiac safety screening of potential new drugs.
Whilst Metrion has a substantial validation dataset for three of the CiPA ‘pillars’ Sarah focused on recent work using Axol’s atrial and ventricular phenotype human iPSC derived cardiomyocytes. Sarah’s presentation can be found here and an accompanying poster, presented at the 2018 Select Biosciences Stem Cells In Drug Discovery meeting, is here. Thanks to all Axol and Metrion staff who contributed towards this work
The pathology of Amyotrophic Lateral Sclerosis (ALS)
The pathology of Amyotrophic Lateral Sclerosis (ALS) was the topic of choice for Gareth Miles from the University of St Andrews, who discussed the use of stem cell-based technology to investigate the interactions between astrocytes and motor neurons in a humanised ALS model. Historic suffers of this disease include American baseball player Lou Gehrig and the renowned physicist Professor Stephen Hawking.
Using iPSC derived co-culture of astrocytes derived from healthy individuals and ALS patients with the TARDPB or C9ORF72 mutations with motor neurones from healthy individuals, the Miles’ group has demonstrated hypo excitability in the ALS astrocyte co-culture motor neurones correlating with loss of sodium and potassium currents. This data, combined with CRISPR studies removing the C9ORF72 ALS-causing mutation, implicates astrocyte-neuron signalling as a promising target for ALS drug discovery.
The phenotypic and functional characterisation of human iPSC derived microglia
Zoe Nilsson from Axol Bioscience then discussed the phenotypic and functional characterisation of human iPSC derived microglia. These are innate immune cells found within the central nervous system which possess key roles in neurogenesis and immunity and which Axol have developed as a co-culture with human iPSC derived cortical neurons. Zoe presented the use of such tools in drug discovery and linked back to Gareth’s earlier talk by describing the dangers of the overactivation of microglia which can lead to neuroinflammation and can play a critical role in ALS and other neurodegenerative disorders.
Using tissue engineering techniques to model Alzheimer’s Disease
Zoe was followed by Eric Hill from Aston University, who described his work using tissue engineering techniques to model Alzheimer’s Disease. Eric spoke about the many difficulties associated with drug discovery in the Alzheimer’s field, attributing this largely to the lack of high quality predictive experimental models. Additionally, it is now widely accepted that the first stages of Alzheimer’s disease may occur around twenty to thirty years before initiation of memory loss – further complicating the situation for disease modelling.
In a quest to produce a high quality predictive in vitro model the Hill lab is pioneering an approach using Alzheimer’s Disease-derived iPSC and 3D culture techniques. As part of the MESO-BRAIN initiative Eric’s team have been 3D printing cultures of Alzheimer’s and healthy’ iPSC astrocytes to form neural networks with defined biological architecture in polymer scaffolds. Conductive polymer scaffolds may be used, enabling monitoring of electrical activity within the organoid structure or, alternatively, real time imaging techniques can be applied. Ultimately the Hill lab aims to produce a model suitable for early discovery compound screening or to trial other novel treatments for Alzheimer’s Disease.
The use of iPSCs in both disease modelling and as a safety pharmacology platform
The final Speaker was Daniel Sinnecker, a cardiologist from the Technical University of Munich. Daniel discussed the use of iPSCs in both disease modelling and as a safety pharmacology platform. For example, use of iPSC as an integral component in CiPA which linked well to earlier content within Sarah Williams’ presentation. Daniel also discussed the use of lentiviral transduction to insert genetically encoded voltage sensors into iPSC cardiomyocytes from healthy and long-QT type 1 (LQT1) patents. Using this technique the Sinnecker lab has been able to quantify cardiomyocyte action potential characteristics in healthy iPSC and also demonstrate early after depolarisations in the LQT1 mutants. This technique shows great promise for evaluating cardiomyocyte iPSC characteristics over an extended time period.
Laboratory demonstrations
After a networking lunch, which prompted further conversation around the various themes presented in the morning session, Metrion Biosciences hosted a series of laboratory demonstrations in our Granta Park facility. The demonstrations involved use of Axol’s iPSCs, with Sarah Williams showcasing Metrion’s “gold standard” conventional manual patch clamp electrophysiology capabilities.
Edd Humphries then demonstrated one of Metrion’s QPatch 48 automated electrophysiology platforms using clonal stable cell lines. The QPatch is a device that produces high quality electrophysiology data for both routine screening in support of medicinal chemistry activities and is the platform upon which Metrion has validated its suite of high quality CiPA-compliant safety profiling assays.
Finally, Said El Haou showcased Metrion’s Axion Biosystem’s Maestro MEA system, a versatile platform able to capture real-time, information rich, recordings from iPSC and cultured native neurons, with the facility to evaluate effects of novel compounds over extended time periods (days to weeks).
Acknowledgements
This event, closely followed the 2018 Cambridge Ion Channel Forum co-hosted by Metrion Biosciences and AstraZeneca on 8th May, was a further example of Metrion Biosciences commitment to promoting and generating high quality science in the Cambridge bio cluster. We would like to thank the Axol team for being excellent co-hosts and we look forward to organising our next event. You can sign up for updates regarding Metrion sponsored events, Metrion’s external speaker presentations and services updates HERE and you can refine your topics of interest and opt out at any time.