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Monday, October 21
 

8:00am

Registration Open
Monday October 21, 2019 8:00am - 6:00pm
Manby Gallery

9:30am

Welcome Coffee in Exhibition
Monday October 21, 2019 9:30am - 10:00am
Manby Gallery

10:00am

10:15am

Keynote Address-OME’s Bio-Formats, OMERO & IDR: Open Tools for Accessing, Integrating, Mining and Publishing Image Data at Scale
Despite significant advances in biological imaging and analysis, major informatics challenges remain unsolved: file formats are proprietary, storage and analysis facilities are lacking, as are standards for sharing image data and results. The Open Microscopy Environment (OME; http://openmicroscopy.org) is an open-source software framework developed to address these challenges. OME releases specifications and software for managing image datasets and integrating them with other scientific data. OME’s Bio-Formats and OMERO are used in thousands of labs worldwide to enable discovery with imaging.

We have used Bio-Formats and OMERO to build solutions for sharing and publishing imaging data. The Image Data Resource (IDR; https://idr.openmicroscopy.org) includes image data linked to >70 independent studies from genetic, RNAi, chemical, localisation and geographic high content screens, super-resolution microscopy, and digital pathology. Datasets range from several GBs to tens of TBs. Wherever possible, we have integrated image data with all relevant experimental, imaging and analytic metadata. With this metadata integration, we have identified gene networks that link to cellular phenotypes. We have also built cloud-based analysis tool portals to catalyse the re-use and re-analysis of published imaging data. Through OME’s commercial arm, Glencoe Software, we have built PathViewer (http://www.glencoesoftware.com/products/pathviewer/), a web-based WSI visualisation and annotation tool that is used in academic medical centres and 10 pharmaceutical companies and for digital pathology data sharing, atlases, analysis, and also for e-learning in medical education.

I’ll present our latest work on linking genotypes and phenotypes in IDR, and our proposals for next generation data formats and public resources for imaging.

Speakers
avatar for Jason Swedlow, Ph.D., FRSE

Jason Swedlow, Ph.D., FRSE

Professor of Quantitative Cell Biology, Centre for Gene Regulation and Expression
Jason Swedlow earned a BA in Chemistry from Brandeis University in 1982 andPhD in Biophysics from UCSF in 1994. After a postdoctoral fellowship with Dr T.J. Mitchison at UCSF and then Harvard Medical School, Dr Swedlow establishedhis own laboratory in 1998 at the Wellcome Trust Biocentre... Read More →


Monday October 21, 2019 10:15am - 11:00am
Wellcome Auditorium

11:00am

11:30am

Enabling Technologies- Deep Learning Techniques for 3D Tissue and Organoid Manipulation and Segmentation
Deep Learning Techniques for 3D Tissue and Organoid Manipulation and Segmentation
In this talk I will give an overview of the techniques developed to convert our high content screening pipeline from 2D to organoids. First, I will present classical image processing algorithms and novel deep learning approaches for single cell-based analysis. I will discuss the challenges, opportunities and difficulties in converting these methods to 3 dimensions. A tool will be presented for annotating volumetric data to generate training image database. Our goal is to develop a deep learning architecture for 3D instance segmentation which will be used for processing image stacks of spheroids acquired on light sheet microscopy. Furthermore, I will present our 3D pipeline from spheroid generation to imaging. We are developing the Spheroid Picker robot that is capable of automatically selecting spheroids based on their morphological properties and transfer them from growing plates to high content plates. This low-cost device is a stereo microscope equipped with a pipette manipulator and a pressure controller system. The clearing method applied on the sample to allow high resolution deep imaging with light sheet fluorescence microscopy will be introduced. Finally, I will describe our label free automatic patch clamp system that performs electrophysiological measurements on living neurons in 3D brain tissue slices. The system is validated on hundreds of rodent and human cells.

Speakers
avatar for Krisztián Koós

Krisztián Koós

Ph.D. Candidate, Biological Research Centre of the Hungarian Academy of Sciences
Krisztian Koos is currently a Ph.D. candidate in the laboratory of Peter Horvath, Biological Research Centre of the Hungarian Academy of Sciences. He graduated as a computer scientist at the University of Szeged where he worked on computer tomography algorithms. His research interests... Read More →

Chair
avatar for Hazel Screen, Ph.D., MRes, BEng

Hazel Screen, Ph.D., MRes, BEng

Professor of Biomedical Engineering, Queen Mary University of London
Hazel Screen is Professor of Biomedical Engineering and Chair of Bioengineering at Queen Mary University of London.Her research group focuses on multi-scale structure-function behaviour and mechanobiology in tissues. She has particular interest in in vitro models to explore health... Read More →


Monday October 21, 2019 11:30am - 11:50pm
Wellcome Auditorium

11:50am

Enabling Technologies- Human Engineered Heart Tissue – An Innovative Model for Preclinical Cardiac Assessment
The presentation will focus on Engineered Heart Tissue technology (EHT). EHTs are three-dimensional, force-generating cardiac tissues in 24 well-format which are generated from single cell suspensions of human induced pluripotent stem cell-derived cardiomyocytes. EHTs are developed between flexible silicone posts under auxotonic stretch. Automated test platforms will be introduced to analyze physiological parameter like contractile force and calcium transients under standardized conditions based on video-optical recording. Specific aspects related to cardiomyocyte maturation and application for in vitro cardiac assessment of compounds will be addressed.

Speakers
AH

Arne Hansen, M.D.

Associate Professor, Department for Experimental Pharmacology and Toxicology, UKE/Hamburg
Arne Hansen received his MD at the University of Hamburg. After clinical training and a 3-year PostDoc at NIH he joined the Department for Experimental Pharmacology and Toxicology at UKE/Hamburg in 2007 and was appointed as Assistant Professor in 2012 and Associate Professor in 2018... Read More →

Chair
avatar for Hazel Screen, Ph.D., MRes, BEng

Hazel Screen, Ph.D., MRes, BEng

Professor of Biomedical Engineering, Queen Mary University of London
Hazel Screen is Professor of Biomedical Engineering and Chair of Bioengineering at Queen Mary University of London.Her research group focuses on multi-scale structure-function behaviour and mechanobiology in tissues. She has particular interest in in vitro models to explore health... Read More →


Monday October 21, 2019 11:50am - 12:10pm
Wellcome Auditorium

12:10pm

Enabling Technologies- Integrated Droplet Microfluidic Platform for 3D Oncology Studies
Spheroids are appealing candidates for replacing 2D cultures since they present a higher level of biological relevance and can be produced at high throughput. Their broad acceptance is hindered by the difficult adaptation to automated screening workflows that have been developed for conventional 2D systems.
In a previous work, we presented an integrated droplet microfluidic chip for the high throughput and reliable production of homogeneous spheroids, for their long term culture and multiscale analysis (Sart, S., Tomasi, R. F.-X., Amselem, G. & Baroud, C. N. Nat. Commun. (2017)). Fluorescent signals and morphological parameters of tens of thousands of spheroids and hundreds of thousands of individual cells were analyzed and correlated in situ. Spheroids were produced in droplets that were trapped in defined locations on chip.

This immobilization allowed easy imaging through time and any perfusion protocol to be performed on chip, like drug perfusion or in situ immuno-cyto-chemistry procedures (including washing steps). More recently we demonstrated the ability to add some content to the droplets at any given time during the experiment through controlled droplet pair merging for controlled 3D co-culture and drug toxicity screening on chip (Tomasi, R. F.-X., Sart, S., Champetier, T. & Baroud, C. N. bioRxiv, (2018)). First droplets were produced, immobilized, and kept in culture for spheroid formation, while second droplets were trapped and merged few days later to bring another cell type or different concentrations of a drug to the initial droplets.

In the present work, this technique is used to perform 3D oncology studies. As the role of the microenvironment is acknowledged to be crucial in tumor progression, we used the sequential microfluidic droplet trapping and merging to build complex 3D tumor cultures. For instance, PC3 cells (prostate cancer) were cultured in 3D with human mesenchymal stem cells that are known to be strong regulators of connective tissues. We are also investigating immunotherapy responses by putting into contact mouse melanoma spheroids (B16-F0) formed after 2 days of cultures and primary mouse cytotoxic T-cells. The microenvironment can also be mimicked by implementing extracellular matrix. For instance we built invasion assays in droplets: after formation of mouse melanoma spheroids in liquid droplets, Matrigel droplets were added and the resulting droplets gelified around each spheroid. One day later, the tumor cells began to invade the matrix.

As cancer progression is inherently associated to tumor growth, we performed long term culture of highly tumorigenic cells in the microfluidic chip. Ewing’s sarcoma spheroids (A673) grew from 60 to 160 µm in diameter within a week a culture in the droplets, each individual being monitored individually by imaging. Finally, the susceptibility of these spheroids to cis-platin was assessed over more than 4 decades of drug concentration on chip.

This technology integrates complex 3D culture protocols on a single microfluidic format, which greatly simplify workflows while benefiting from the miniaturization (higher density, cost savings, low sample volumes). By providing reproducible complex 3D environments at high throughput, this technology could have a strong impact in oncology, drug discovery and personalized medicine.

Co-Authors:
Aude DURAND, Pasteur
Aimee WESSEL, Pasteur
Gustave RONTEIX, Pasteur
Christelle ANGELY, Pasteu
Elaine DEL NERY, Curie
Charles BAROUD, Pasteur

Speakers
avatar for Raphael Tomasi

Raphael Tomasi

Post Doctoral Researcher, Ecole Polytechnique Institut Pasteur
Raphaël TOMASI graduated from the ESPCI Paris in 2013 with an engineering degree as well as a research master in microfluidics. He gained his PhD from the hydrodynamics laboratory (LadHyX) of the Ecole Polytechnique (Paris area) in 2016, under the supervision of Charles BAROUD. Raphaël... Read More →

Chair
avatar for Hazel Screen, Ph.D., MRes, BEng

Hazel Screen, Ph.D., MRes, BEng

Professor of Biomedical Engineering, Queen Mary University of London
Hazel Screen is Professor of Biomedical Engineering and Chair of Bioengineering at Queen Mary University of London.Her research group focuses on multi-scale structure-function behaviour and mechanobiology in tissues. She has particular interest in in vitro models to explore health... Read More →


Monday October 21, 2019 12:10pm - 12:30pm
Wellcome Auditorium

12:30pm

Enabling Technologies- Multiparametric Phenotyping of Compound Effects on Patient Derived Organoids
Patient derived organoids (PDOs) closely resemble individual tumor biology and allow testing of small molecules ex vivo. To systematically dissect compound effects on 3D organoids, we developed a high-throughput imaging and quantitative analysis approach. We generated PDOs from colorectal cancer patients, treated them with >500 small molecules and captured >3 million images by confocal microscopy. We developed the software framework SCOPE to measure compound induced re-organization of PDOs. We found diverse, but re-occurring phenotypes that clustered by compound mode-of-action. Complex phenotypes were not congruent with PDO viability and many were specific to subsets of PDO lines or were influenced by recurrent mutations. We further analyzed specific phenotypes induced by compound classes and found GSK3 inhibitors to disassemble PDOs via focal adhesion signaling or that MEK inhibition led to bloating of PDOs by enhancing of stemness. Finally, by viability classification, we show heterogeneous susceptibilities of PDOs to clinical anticancer drugs.

Authors:
Johannes Betge, Niklas Rindtorff, Jan Sauer, Benedikt Rauscher, Clara Dingert, Haristi Gaitantzi,
Frank Herweck, Thilo Miersch, Erica Valentini, Veronika Hauber, Tobias Gutting, Larissa Frank, Sebastian Belle,
Timo Gaiser, Inga Buchholz, Ralf Jesenofsky, Nicolai Härtel, Tianzuo Zhan, Bernd Fischer, Katja Breitkopf-Heinlein, Elke Burgermeister, Matthias P. Ebert, Michael Boutros

Speakers
avatar for Niklas Rindtorff

Niklas Rindtorff

MD/Ph.D. Student, German Cancer Research Center (DKFZ)

Chair
avatar for Hazel Screen, Ph.D., MRes, BEng

Hazel Screen, Ph.D., MRes, BEng

Professor of Biomedical Engineering, Queen Mary University of London
Hazel Screen is Professor of Biomedical Engineering and Chair of Bioengineering at Queen Mary University of London.Her research group focuses on multi-scale structure-function behaviour and mechanobiology in tissues. She has particular interest in in vitro models to explore health... Read More →


Monday October 21, 2019 12:30pm - 12:50pm
Wellcome Auditorium

12:50pm

1:00pm

2:30pm

Poster Presentation #1- High-Throughput Drug Screening with Advanced Cell-Based Assays
High-Throughput drug screening with advanced cell-based assays

Booij T.H., Hirt C., Keller D., Taube D., Prummer M., Riegler E.M., Stirnimann C., Schwank G.

Lab automation has enabled miniaturization and high-throughput screening approaches in drug research, allowing increased numbers of potential drugs to be tested in in vitro assays. Screens have largely been performed using simple 2D cell culture assays to minimize assay variability. However, it is increasingly being recognized that simple 2D cell culture assays may not adequately represent human disease, causing poor translatability from in vitro results to in vivo experiments. As a result, more advanced assays have gained popularity, such as 3D cell culture assays with cultured cell lines, or the use of patient material as PDX- or organoid models. While the use of such assays has, initially, largely been limited to neoplastic disorders such as cancer and cystic diseases, these models are gaining in popularity for other indications, especially where in vitro experimentation has not been general practice.

A limitation of more advanced cell-based assays with PDX- or organoid model systems has been to automate the liquid-handling procedures to enable high-throughput drug screening in these models. The main factors holding back this development include reagent costs, difficulties with liquid handling of viscous reagents, and the increased assay variation that is inherent to the growth of ‘mini-organs’ in normal tissue culture.
We have established a statistically robust organoid screening platform for pancreatic cancer that integrates fully-automated liquid handling, plate transfer and readouts. This screening platform was applied to screen a drug library that can allow for rapid drug repurposing, possibly providing new lead compounds for this, so far, largely incurable disease.

Speakers
DK

David Keller

Lab Technician, ETH Zurich NEXUS Personalized Health Technologies


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #10- Phenotypic Screening in 3D Culture Including the Extracellular Matrix
Ninety percent of potential drug candidates fail within the clinical trials, mainly because of a lack of efficacy. Biologists and pharmacologists now largely acknowledge the lack of relevant in vitro models in the early drug development stages.

To mimic human tissues, several parameters need to be taken into account: the target cells but also the extra-cellular matrix (ECM). The ECM has diverse and crucial roles as anchorage of the cells, migration barrier and track, signal reservoir, low-affinity co-receptor, signal presenter and is a source of biochemical forces. All these roles of the matricial micro-environment are really important in maintaining organ’s homeostasis and proper intercellular communications. Depending on the organ, ECM composition and physicochemical properties are different.

Cellular assays in 3D culture have shown many advantages to better mimic in vivo situation. Nevertheless, 3D technology used for phenotypic screening is often really simple, as the use of ULA plates. But we can go further by using 3D technologies, which allow reproducing the microenvironment of healthy or pathological organ of interest, as BIOMIMESYS technology. During this talk, we will see how the cellular microenvironment impacts the proliferation and/or differentiation of cells. Few examples will be presented in oncology, CNS and metabolic diseases fields.

Speakers
NM

Nathalie Maubon, Ph.D.

President, HCS Pharma
With 22 years of experience in the pharmaceutical research (public research, pharmaceutical industry and biotechnology company), Nathalie Maubon is a specialist in the development and automation of robust models for high-throughput screening (HCS, HTS, ADMET). She has held several... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #2- Time-Lapse Imaging of Cell Behaviour on 2D Substrates and in 3D Aggregates: Watch it Live with DRAQ9
The importance of cell migration and proliferation in wound healing, tumour metastasis and tissue remodelling is under continued investigation while the ability to track single cells and cellular communities has become a potent tool to measure their biological functional capacity and a systems response to perturbation. Delivering a tracking capability and robust assay readouts is multi-faceted. It requires a cell label that is non-toxic, stable throughout, as cell identifier/locator. We describe a cell permeant, far-red fluorescent, cytoplasmic probe DRAQ9 (BioStatus) that is non-toxic, and present continuously as real-time tracker so fluorescence is undiminished over time e.g. during spheroid aggregation, formation and expansion; in 2D scratch-wounds; or a hybrid assay, as single cells emerge onto 2D substrate from a spheroid. In all cases, DRAQ9 labels cells for unique identification over time. This stable tracker provides a readout ripe for automated cell tracking in aggregates, migration in 2D or indeed through mitosis.

Additionally, in end-point analyses (e.g. high throughput toxicity assays) a fundamental first step to 3D object image analysis is correct boundary identification, where failure results in inaccurate downstream morphometric analysis. DRAQ9 functions as a whole 3D-object “paint”. This allows accurate reproducible boundary identification, using an inexpensive universal platform, simplifying image analysis, for rapid, effective screening.

A459 cells were cultured in wells using a 2 Well insert (IBIDI). After 24 h inserts were removed, cells washed and DMEM (with 10% FBS, 2 μM DRAQ9) added then returned to the incubator for 20 min. Time-series images were acquired every 20 min. using 20x objective lens, for 24 h. Cells were manually tracked and analysed using imageJ. Spheroids were generated in a U-bottomed low-adherence plate from A549 cells cultured in DMEM (with 2 μM DRAQ9, upon spheroid culture) for 7 days. Time-series images were acquired every 30 min. using 10x objective lens, for 4 days. After 7 days spheroids were re-plated to adhere for outgrowth and time-series images acquired every 20 min. using 10x objective lens, for 48 h. A widefield microscope was used throughout.

Manual cell tracking can be completed with confidence due to DRAQ9's cytoplasmic localisation and absence in nuclei. Distinct cell patterning enables automated cell tracking algorithms. Mitosis can be visualised from changes in DRAQ9 signal due to altered morphology. Spheroid painting allows confident morphometric analysis of 3D cultures over time due to non-toxic properties of DRAQ9 for time-series analysis of cultures. Emerging cell behaviour can be visualised from 3D cultures.

DRAQ9 labeling of live cell cultures enables real-time tracking of cell migration and cell division. Visualisation of cell-cell and cell-matrix interactions provides rich information on cell behaviour within a niche. Far-red fluorescence makes it compatible with imaging on sub-optimal imaging substrates e.g. tissue culture plastic. Changes in fluorescence intensity adds information on cellular division, allowing such events to be localised to a cell niche. DRAQ9-brightfield image overlays make manual tracking easier to perform, increasing confidence in cell identification, especially during cell division. Importantly, absence of DRAQ9 in nuclei provides a mask for automated image analysis, reducing image analysis times.

Speakers
RE

Roy Edward, BSc Hons

Sales and Marketing Director, BioStatus Limited
Trained in Biochemistry (St Andrews Univ.). 30+ years of international experience in biotools, in biomagnetic separation and proteomics.Beyond the UK, worked in the USA and Norway with roles in product and marketing management, international distributor management and business development.Written... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #3- Magnetic Electrospun Microfibre Scaffold Assemblies: Examples of there use for Cell-Based Screening Applications
In an effort to create a more physiological three-dimensional (3-D) environment for cell growth while maintaining the logistics of well plate screening for drug discovery purposes, we have develop a 3-D micro-scaffold platform from electrospun material used in conjunction with well plates for higher throughput screening. We have engineered electrospun material to form micro-scaffold islands of cells. Cells grow on, around and into the material, forming a micro-island of adherent cells that are effectively 3-D in solution. The incorporation of iron nano-particles into the fibres during manufacture results in micro-scaffold islands that can be physically manipulated using electro-magnetism. We can culture cells on micro-scaffolds in culture vessels in media within the incubator in addition to moving them within wells using arrays of neodymium magnets, attracting scaffolds to the edges of wells to prevent damage during media replacement or align scaffolds in the centre of wells for imaging purposes. Using sheathed electromagnetic tips we can move micro-scaffolds from well to well with no adverse effects to cells. In addition we have developed a liquid handling device capable of identifying the location of scaffolds in a petri dish then performing a ˜pick and place™ procedure putting them into user defined wells of a plates. Furthermore, we have incorporated fluorescent dyes or quantum dots into the fibres such that fibres can be visualised using fluorescence microscopy, while quantum dots can be used as a barcode to distinguish between two cell populations on different scaffolds within the same well. Cells can be transfected while cultured on micro-scaffolds using either lipofection or electroporation. Recombinant cells can be cryo-preserved on micro-scaffolds. We will show data of exemplar assays using recombinant, primary and differentiated stem cell assays on micro-scaffolds. We have used this approach to examine a number of luminescence and fluorescence readouts in recombinant cell lines and have differentiated iPSC™s to cortical neurones in 3-D on micro-scaffolds to characterise then use imaging as a readout. Our data suggests that this micro-scaffold approach may open up new opportunities for both recombinant cell based screening and differentiated stem cell assays using single and co-cultures in a well plate-based format more amenable to higher throughputs with very little manual intervention.

Speakers
GA

Gary Allenby, Ph.D.

Chief Scientific Officer, Aurelia Bioscience
PhD in reproductive tox, worked in pre-clinical pharma for 25 years in Hit ID, Hits to Lead, Target Biology sections developing cell based assays for compound screening. Founding entrepreneur of Aurelia Bioscience in 2012, delivering tailored assay development for compounds pharmacological... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #4- 3D Multicellular Intestine-on-a-chip Model for Disease Modeling and Drug Discovery
The development of efficient drugs and therapies for treatment of inflammatory bowel disease (IBD) is hampered because of the lack of reliable in vitro and in vivo models. Current models generally fail to recapitulate key aspects of the intestine, resulting in low translatability to the human situation. With the increasing IBD incidence worldwide, there is an unmet need for predictive in vitro models that recapitulate the complex structure of the human intestine. Here, we show the development and characterization of a 3D multicellular perfused intestine-on-a-chip model and its application for investigating and interfering with intestinal inflammation.

Forty independent polarized 3D perfused leak-tight epithelial tubular structures composed of Caco-2 and HT29-MTX-E12 cells were obtained in the microfluidic platform, the OrganoPlate 3-lane. Next, immune competent cells were added to the system and shown to be functional, as demonstrated by the release of cytokines upon activation.

To mimic intestinal inflammation, the model was exposed to tumor necrosis factor (TNF) α and/or interleukin (IL)-1β, whose effects were assessed by measuring the transepithelial electrical resistance (TEER) and secretion of proinflammatory cytokines at the apical and basal sides. The cytokines induced an inflammatory state in the culture, as demonstrated by the impaired barrier integrity and increased IL-8 secretion. We also assessed the applicability of the model in screening anti-inflammatory compounds by using the well-known anti-inflammatory drug TPCA-1. Exposure to the drug prevented an inflammatory state in the model.

Overall, these results suggest that this model provides the versatile modularity of mimicking key features of intestinal inflammation, which positions it at the forefront of high-throughput screening efforts for supporting drug discovery and providing a platform for personalized medicine.

Speakers
LG

Linda Gijzen

Scientist, Mimetas BV
Linda Gijzen, MsC, got her master’s degree in Toxicology and Pathobiology from the Radboud University, the Netherlands, in 2016. Since then she is working as a scientist at the Model Development department at Mimetas, the organ-on-a-chip company, in Leiden. There she is involved... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #5- Patient-Derived Explants (PDEs) as a Powerful Ex Xivo Platform to Predict Clinical Response to Novel Anti-Cancer Drugs
One of the leading goals of the Leicester Cancer Research Centre is to investigate novel and more relevant pre-clinical tumour models that are able to better predict patient outcomes.

Accordingly, we have established a rapidly expanding ‘Cancer Explant Facility’ whereby Patient-Derived Explants (PDEs) from freshly resected tumour specimens, are cultured ex vivo and tested with a wide panel of anti-cancer drugs including currently used therapies as well as novel drugs. This ex vivo model is a powerful platform with multiple advantages over animal models: it is patient-specific, it improves the predictivity of drug response in situ and, by accurately retaining the complexity of the tumour ex vivo, overcomes the differences between the human and animal immune systems that can lead to differential response to targeted immune-oncology therapies in animal models.

In our experimental method, drug responses are assessed using multiplexed immunofluorescence (mIF), which allows the ability to distinguish between drug effects in tumour areas versus the surrounding stroma as well as the potential contribution of the host immune system. So far, this fast and reproducible technique has been applied to different types of cancer (melanoma, lung, renal, breast and CRC liver metastasis) to analyse the effect of anti-cancer agents on cell viability, proliferation and immune cell mobility. Finally, by combining this platform with gene expression, genetic and epigenetic analysis, we also aim to identify novel biomarkers that correlate with, and predict, the response of cancers to anti-cancer drugs as well as their toxicity in preclinical settings, prior to clinical testing.

Speakers
GV

Giuditta Viticchie, PhD

Post Doctoral Research Associate, University of Leicester
2008: Master’s degree in Chemistry at the University of Rome “Tor Vergata”, Italy.2012: PhD in Biochemistry and Molecular Biology at the University of Rome “Tor Vergata”, Italy2012-2013: Post Doctoral Fellow at the MRC Toxicology Unit Leicester, UK, with Italian fellowship... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #6-Real-Time Live-Cell Analysis of 3D Tumour Spheroid Invasion
Cell invasion, a hallmark of malignant cancers, plays a key role in tumour dissemination and metastasis and is responsible for the higher incidence of cancer-related deaths. The ability of tumour cells to form a metastatic tumour primarily involves cell morphological reorganization and degradation of the surrounding extracellular matrix (ECM).Conventional in vitro invasion models, such as the filter based transwell invasion assay (2D) are widely used to assess tumour cell invasion.However, lack of comprehensive cell-ECM interactions may limit their physiological relevance. Here, we provide a simple 3D tumour spheroid invasion model to recapitulate key features of the metastatic phenotype and demonstrate use of live cell imaging to kinetically acquire and quantify 3D single spheroid invasion.



To exemplify the assay platform and assess the metastatic potential of tumour cells in 3D, U87-MG, A172 and HT1080 spheroids were formed for 72 h in Ultra-Low Attachment round bottom 96-well plates and subsequently embedded in Matrigel (4.5 mg/mL). Spheroid invasive properties were measured using novel brightfield image analysis software (IncuCyte® Live-Cell Analysis System) which tracks and quantifies changes in spheroid size (whole spheroid area or invading cell area) over time. To account for variation in size following spheroid formation, whole spheroid area was normalized to spheroid size at t = 0 h and automatically plotted. Cell type specific temporal invasive potential was observed. At 168 h, whole spheroid area of U87-MG spheroids (ratio: 32) increased to approximately 2x and 4x the size of HT1080 (ratio: 18) and A172 (ratio: 8) spheroids respectively. HT1080 spheroids embedded in varying concentrations of Matrigel (1.13 – 4.5 mg/mL) demonstrated that spheroid invasive capacity is Matrigel concentration-dependent. While cell motility was reduced at the highest Matrigel concentration, marked density and elongation of invadopodia was seen. Using available metrics and analysis approaches we could determine the effects of known anti-metastatic compounds on spheroid invasion and spheroid proliferation. Cytochalasin D (300 nM) and PP242 (30 µM) inhibitors caused comparable reduction of U87-MG spheroid size (90% and 80% respectively). However, marked inhibition of invadopodia was only evident with Cytochalasin D. Subtraction of the invading cell area from the whole spheroid area provided a measure of the spheroid body size and revealed the anti-proliferative rather than anti-invasive properties of PP242. A separate but identical study conducted in the absence of Matrigel further supported the anti-proliferative effects of PP242 but not Cytochalasin D.

These data demonstrate the capability to kinetically visualise and quantify 3D single spheroid invasion assays and illustrates the utility of this approach for high throughput real-time anti-metastatic compound profiling. This model can be further developed with the incorporation of stromal cells to address additional aspects of tumour cell invasion.

Speakers
MO

Miniver Oliver

Research Scientist, Sartorius
Research Scientist at Sartorius, working as part of the Applications group for the IncuCyte platform. Expertise indeveloping 3D cellular assays for quantitative live cell analysis.


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #7- High-Content Assay for Morphological Characterization of 3D Neuronal Networks in a Microfluidic Platform
Establishment of physiologically-relevant in vitro models is crucial to further understanding of the mechanisms of neurological diseases as well as targeted drug development. While iPSC-derived neurons show great promise for compound screening and disease modeling, use of three-dimensional (3D) cultures is emerging as a valid approach for neuronal cell based assay development. 3D cultures are recognized as more closely recapitulating aspects of the human tissues including the architecture, cell organization, as well as cell-cell and cell-matrix interactions.

This poster presents the development of a high-throughput 3D neurite outgrowth assay using iPSC-derived neurons developed in the microfluidic OrganoPlate® platform, with the goal of establishing 3D models for neurodegenerative diseases and neurotoxicology screens. 3D cultures were treated with known neurotoxic compounds and the morphology and viability of neurons were assessed. The OrganoPlate® from MIMETAS is a high-throughput platform that combines the most recent advances in 3D cell culture, Phaseguides™ and microfluidics. The OrganoPlate contains 96 tissue chips suitable for long term culture of live cells, is amenable for screening purposes, and is compatible with standard laboratory equipment or automated systems, like the ImageXpress® Micro Confocal High-Content Imaging System.

Speakers
AB

Andy Bashford

Application Scientist, Molecular Devices
Andy works as a Field Applications Scientist for Molecular Devices covering the whole portfolio of imaging solutions.Prior to this he worked as a development scientist for the European Collection of Authenticated Cell Cultures (ECACC) where he was responsible for developing assays... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #8- LaFOS: A Novel 4D Morphometric Phenotypic Profiling Method to Uncover Therapeutic Vulnerabilities in Patient-Derived 3D Tumor Models
3D-spheroid cultures are powerful tools for in vitro models for biology, but size and shape diversity within the culture is largely ignored and also their current end point data outputs (like metabolic measurements) are resource intensive and we cannot afford real-time kinetics at the primary drug screening level. We intent to develop robust Label free Oncology Score (LaFOS) using brightfield image of 3D tumor spheroids in different time intervals. We developed a computational image scoring methodology, that integrates morphometric profiling, segmentation, filtering, and analysis for brightfield images of 3D culture and provide us with cytotoxicity level without any metabolic end point readouts like cell viability assay. LaFOS is accurate and flexible like current existing platforms, and we illustrate its potential by studying 600 plus small molecules that are used in the clinic or in advanced phases of development. We tested this computational method in parallel with cell viability assay (cell titer Glo) in 15 primary patient derived models and 3 commercial cancer cell lines in comparison with traditional 2D monolayer models. We found Pearson correlation of 0.81 for LaFOS methodology and 0.95 for cell viability assay in finding the cellular cytotoxicity. Our results suggest that, there is differential drug response between 2D and 3D model for specific class of compounds. The compound connectivity map showed different targets specificity and network structure between 2D and 3D models. We observed pathway specific enrichment of (EGFR, PI3K and mTOR inhibitors) in 3D models and cell cycle/division related enrichment (HDAC, CDK, Topo and Tubulin) in 2D models. RNA-sequence analysis of 13 cell lines showed 151 pan-differentially expressed genes in 2D vs 3D. Immunofluorescence and western blot analysis showed HIF-1α was high in 3D models and Ki67 was found high in 2D models this data correlates to architecture dependent drug response and gene expression data. Also 3D models showed more activity of pAKT and pERK could be a reason for pathway specific inhibitor activity in 3D models. We did a validation of hits obtained from in vitro screen using in vivo PDX model for ZSTK474 a PI3K inhibitor. Overall we developed a computational platform (LaFOS) that can measure 3D spheroid cell viability in real-time fashion using morphometric signature obtained from label free brightfield phenotypes.

Speakers
GP

Giridharan Periyasamy, PhD

Platform Leader, Genome Institute of Singapore
Dr. Giridharan Periyasamy is a Platform Leader in Centre for High Throughput Phenomics (CHiP-GIS) at the Genome institute of Singapore. He is the domain expert in Assay automation, miniaturization, robotics, and High Content Imaging technology. His research focus is on the Cancer... Read More →


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

2:30pm

Poster Presentation #9- Microfluidic Drug Efficacy Testing of 3D Tumour Models
In recent years, physiologically relevant 3D in vitro models of cancer have been continuously refined in order to better mimic the tumour microenvironment when testing chemotherapies and immunotherapies. An increasing interest in precision treatment of cancer patients has highlighted the need for technologies that are capable of maximising the testing of compounds in 3D tumour models obtained from cancer biopsies. Microfluidics and organ-on-a-chip technologies, via precise control of fluids and cells in solution, offer great opportunities for screening biopsy-derived spheroids, organoids, co-cultures and primary tumour fragments in a sophisticated and cost-effective manner.

We have developed microfluidic platforms for high quality and multiplexed drug screening assays on both spheroid, organoids and tissue fragments. Readouts, such as the spheroid volume, spheroid shape and viability are generated from both brightfield and epifluorescence microscopy. The technology has been validated using a variety of cell lines, primary and biopsy cancerous tissue. As proof of concept, human prostate biopsies were grown as a heterogeneous co-culture and used for the generation and screening of thousands of spheroids, showing sensitivity to docetaxel, but resistance to enzalutamide, despite the presence of intact androgen receptors. A further application of the platform lies in the co-culture of several cell types, in order to reconstruct a more realistic tumour microenvironment. Several breast cancer cell lines and cancer-associated fibroblasts (CAFs) were tested to allow the assessment of the impact of drugs and inhibitors on specific cell populations. Lastly, ovarian tumour biopsy tissue fragments were cultured for precision medicine purposes.

Speakers
TM

Theresa Mulholland, BsC

Senior Scientist, ScreenIn3D Limited
Senior scientist at ScreenIn3D, interested in the development of novel assays for drug development and screening, using complex microfluidic 3D in vitro models


Monday October 21, 2019 2:30pm - 3:00pm
Sherry Coutu Seminar Suite Foyer

3:00pm

Lost in Translation: Success through Collaboration-The Organ on a Chip Technologies Network
The Organ-on-a-Chip Technologies Network is a UKRI funded Technology Touching Life initiative, designed to capture, inspire and grow UK research activity in the Organ-on-a-Chip research field. Our global aims are to:
•Develop a vibrant multidisciplinary research community, bringing focus to the varied Organ-on-a-Chip and in vitro model research activity in the UK;
•Facilitate inter-disciplinary and inter-sectoral research collaborations, to develop the next generation of organ-on-a-chip research solutions;
•Train, support and inspire the next generation of outstanding leaders in organ-on-a-chip research.

Our flagship sabbatical funding programme is designed to pump prime new collaborations and research activity in the field. Researchers can spend time in a different (cross-discipline) laboratory. Funding includes salary for the sabbatical period plus up to £5k consumables and a travel/subsistence grant.
We run 6 monthly network events, which aim at sharing members latest research, facilitating new collaborations and identifying routes to support our community. We are currently establishing a range of special interest groups within the network, providing financial support for groups to run more focused organ-on-a-chip workshops and events. We also provide funding for network members to travel between each other’s laboratories to initiate new collaborations.

We have a strong focus on early career researcher support and training, running activities specifically for this group. Currently, our early career researchers are spearheading new public engagement activities for the organ-on-a-chip community with support (financial, training and delivery) from our team.
The network is free and simple to join: https://www.organonachip.org.uk/

Members of our leadership team can provide further information:
Hazel Screen (QMUL)
Martin Knight (QMUL)
Anthony Bull (Imperial Col)
Alicia El Haj (Birmingham)
Andy Carr (Oxford)
Matt Dalby (Glasgow)
Katia Karalis (Emulate)
Paul Workman (ICR)

Speakers
avatar for Hazel Screen, Ph.D., MRes, BEng

Hazel Screen, Ph.D., MRes, BEng

Professor of Biomedical Engineering, Queen Mary University of London
Hazel Screen is Professor of Biomedical Engineering and Chair of Bioengineering at Queen Mary University of London.Her research group focuses on multi-scale structure-function behaviour and mechanobiology in tissues. She has particular interest in in vitro models to explore health... Read More →

Chair

Monday October 21, 2019 3:00pm - 3:15pm
Wellcome Auditorium

3:15pm

Lost in Translation: Success through Collaboration- Utilizing 3-Dimensional Functional Genomics Screening to Identify CREBBP as a Novel Tumour Suppressor in Breast Cancer
Rationale - The contribution of the majority of frequently mutated genes to tumourigenesis is not fully defined. Many aggressive human cancers, such as triple negative breast cancers (TNBCs), have a poor prognosis and lack tractable biomarkers and targeted therapeutic options.

Methods - Here, we systematically characterize loss-of-function mutations to generate a functional map of novel driver genes in a 3-dimensional (3D) model of breast cancer heterogeneity that more readily recapitulates the unfavourable tumour microenvironment in vivo i.e. nutrient stress.

Results - We identified several genes, including CREBBP, FOXA1, KMT2C and NIPBL, whose silencing provided a 3D specific growth advantage. Indeed, the histone acetyltransferase CREBBP was a potent tumour suppressor gene whose silencing provided a 3D-specific growth advantage only under oxygen and nutrient deplete conditions.

CREBBP protein expression was altered in a third of TNBCs as well as several other solid tumours, including bladder, ovarian and squamous lung cancers. In multiple primary tumours and cell models, loss of CREBBP activity resulted in upregulation of the FOXM1 transcriptional network. Strikingly, treatment with a range of CDK4/6 inhibitors (CDK4/6i), that indirectly target FOXM1 activity, selectively impaired growth in both CREBBP-altered spheroids, patient-derived organoids and cell line xenografts from multiple tumour types.

Conclusions - This study is the first to provide rationale for CREBBP as a biomarker for CDK4/6i response in cancer representing a new treatment paradigm for tumours that harbour CREBBP alterations that have limited therapeutic options.

Authors - Barrie Peck, Philip Bland, Ioanna Mavrommati, Hannah Cottom, Patty T Wai, Sarah L Maguire, Holly E Barker, Eamonn Morrison, Divya Kriplani, Lu Yu, Amy Gibson, Giulia Falgari, Keith Brennan, Gillian Farnie, Rebecca Marlow, Daniela Kolarevic, Snezana Susnjar, Natasa Medic Milijic, Kalnisha Naidoo, Patrycja Gazinska, Ioannis Roxanis, Sunil Pancholi, Lesley-Ann Martin, Erle M Holgersen, Maggie CU Cheang, Farzana Noor, Sophie Postel-Vinay, Gerard Quinn, Simon McDade, Lukas Krasny, Paul Huang, Frances Daley, Gareth Muirhead, Syed Haider, Fredrik Wallberg, Jyoti S. Choudhary, Andrew N Tutt, & Rachael Natrajan

Speakers
avatar for Barrie	Peck, Ph.D.

Barrie Peck, Ph.D.

Group Leader, Barts Cancer Institute
I completed my Ph.D. in breast cancer signalling at Imperial College London in 2010. I then moved to the London Research Institute to the lab of Prof. Almut Schulze to study cancer metabolism. This work focussed on identifying novel cancer-specific dependencies in cancer metabolism... Read More →

Chair

Monday October 21, 2019 3:15pm - 3:25pm
Wellcome Auditorium

3:25pm

Lost in Translation: Success through Collaboration- Magnetic Electrospun Microfibre Scaffold Assemblies: Examples of Their Use for Cell-Based Screening Applications
In an effort to create a more physiological three-dimensional (3-D) environment for cell growth while maintaining the logistics of well plate screening for drug discovery purposes, we have develop a 3-D micro-scaffold platform from electrospun material used in conjunction with well plates for higher throughput screening. We have engineered electrospun material to form micro-scaffold islands of cells. Cells grow on, around and into the material, forming a micro-island of adherent cells that are effectively 3-D in solution. The incorporation of iron nano-particles into the fibres during manufacture results in micro-scaffold islands that can be physically manipulated using electro-magnetism. We can culture cells on micro-scaffolds in culture vessels in media within the incubator in addition to moving them within wells using arrays of neodymium magnets, attracting scaffolds to the edges of wells to prevent damage during media replacement or align scaffolds in the centre of wells for imaging purposes. Using sheathed electromagnetic tips we can move micro-scaffolds from well to well with no adverse effects to cells. In addition we have developed a liquid handling device capable of identifying the location of scaffolds in a petri dish then performing a ˜pick and place™ procedure putting them into user defined wells of a plates. Furthermore, we have incorporated fluorescent dyes or quantum dots into the fibres such that fibres can be visualised using fluorescence microscopy, while quantum dots can be used as a barcode to distinguish between two cell populations on different scaffolds within the same well. Cells can be transfected while cultured on micro-scaffolds using either lipofection or electroporation. Recombinant cells can be cryo-preserved on micro-scaffolds. We will show data of exemplar assays using recombinant, primary and differentiated stem cell assays on micro-scaffolds. We have used this approach to examine a number of luminescence and fluorescence readouts in recombinant cell lines and have differentiated iPSC™s to cortical neurones in 3-D on micro-scaffolds to characterise then use imaging as a readout. Our data suggests that this micro-scaffold approach may open up new opportunities for both recombinant cell based screening and differentiated stem cell assays using single and co-cultures in a well plate-based format more amenable to higher throughputs with very little manual intervention.


Speakers
GA

Gary Allenby, Ph.D.

Chief Scientific Officer, Aurelia Bioscience
PhD in reproductive tox, worked in pre-clinical pharma for 25 years in Hit ID, Hits to Lead, Target Biology sections developing cell based assays for compound screening. Founding entrepreneur of Aurelia Bioscience in 2012, delivering tailored assay development for compounds pharmacological... Read More →

Chair

Monday October 21, 2019 3:25pm - 3:35pm
Wellcome Auditorium

3:35pm

Lost in Translation: Success through Collaboration- Magnetic 3D Bioprinting, from Spheroids to Fingerprinting Cells in 3D Using a 2D Workflow
The growing push for 3D cell culture models is limited by technical challenges in handling, processing, and scalability to high-throughput applications. To meet these challenges, we use our platform, magnetic 3D bioprinting, in which cells are individually magnetized and assembled with magnetic forces. In magnetizing cells, not only do we make routine cell culture and experiments feasible and scalable, but we also gain fine spatial control in the formation of spheroids and more complex structures. This presentation will focus on recent developments using this platform, particularly in cancer biology and immunology. Specifically, we will present a method for phenotypic profiling of cell types within spheroids using real-time high-throughput imaging. This label-free method allows for multiplexing with other assay endpoints for high-content screening. Overall, we use magnetic 3D bioprinting to create functionally and structurally representative spheroids for high-throughput screening. 

Speakers
GS

Glauco Souza, Ph.D.

Director of Global Business Development and Innovation and Assistant Adjunct Professor, Greiner Bio-One and University of Texas Health Science Center at Houston

Chair

Monday October 21, 2019 3:35pm - 3:45pm
Wellcome Auditorium

3:45pm

Lost in Translation: Success through Collaboration- A Scalable iPSC-Derived Blood Brain Barrier Penetration Assay in Perfused Endothelial Micro-Tubes to Test Anti-Inflammatory Drugs
The human blood–brain barrier (BBB) is a protective and regulatory interface that permits entry of essential nutrients, while preventing harmful substances from entering the central nervous system (CNS). From the pharmacological perspective, the BBB is one of the major hurdles of CNS drug delivery. Recent BBB models based on human iPSC-derived brain microvascular endothelial cells (BMECs) provide optimal BBB properties and scalability for brain-penetration tests in early drug discovery. However, advanced BMEC differentiation protocols require expensive co-cultures of primary cells to improve the BBB phenotype and thereby hamper the upscaling of a screen. Here we present a contemporary, cost effective and scalable BMEC differentiation method for brain-penetration tests with primary cell-conditioned media that provide similar barrier properties compared to co-culture methods. In a pilot study, a collection of anti-inflammatory compounds have been tested for brain-penetration compared to reference substances. After confirming the integrity of the endothelial barrier, the concentrations of each permeated compound was measured. Supernatants containing passed compounds were finally used to quantify their anti-inflammatory effect in a cytokine-release assay based on human immune cells. In summary, toxicity, penetrability, and anti-inflammatory functionality were determined for each compound. Finally, our method was miniaturized on a filter-free organ-on-a-chip platform allowing tube-like formation of matured BMECs in a perfused microfluidic device which reflects in vivo BBB physiology even better. 

Speakers
avatar for Sven Fengler, Ph.D.

Sven Fengler, Ph.D.

PostDoc, DZNE Bonn Germany
2017 - current, PostDoc Laboratory Automation Technologies (LAT) – Core Research Facilities & Services, German Centre for Neurodegenerative Diseases (DZNE), Bonn, Germany 2015 - 2017, PostDoc Department of Molecular Biology I, Centre for Medical Biotechnology (ZMB) University Essen... Read More →

Chair

Monday October 21, 2019 3:45pm - 3:55pm
Wellcome Auditorium

3:55pm

Lost in Translation: Success through Collaboration- Robotic Lab Assistant for Automated Microfluidic 3D Microtissue Production
We present a general automation platform for droplet microfluidics with a conversational mobile interface. We demonstrate fully automated production of standardized human cancer 3D microtissues from OvCar8, RT4, BJhTERT, RT4-BJhTERT co-cultures, J82, TCCSUP, TYK-nu, HEK, CHO cells as well as primary human hepatocytes, at a throughput of 85000 spheroids per microfluidic circuit per hour using this platform. Spheroids are automatically assembled, incubated and retrieved with high viability. HEK cell viability decreased by only 4%, from 96% after spheroid formation and retrieval. The platform interfaces with standard labware. This allows the scientist to perform further downstream analysis on the spheroids. 45 μm spheroids continue to grow on plates for 5 days, reaching 135 μm in diameter. Finally, RT4 (bladder cancer)-BJhTERT (fibroblast) co-culture spheroids prepared by the robotic assistant were tested against Gemcitabine as a model for future automated workflows for precision oncology. The automated large scale production of engineered 3D microtissues is part of a larger effort to implement a workflow for near-patient drug development. 

Speakers
HN

Haakan N Joensson, Ph.D.

Associate Professor, KTH Royal Institute of Technology Div of Protein Science SciLifeLab
KTH Nanobiotechnology (Alfa4), Science for Life Laboratory, Box 103117121 Solna, Sweden

Chair

Monday October 21, 2019 3:55pm - 4:05pm
Wellcome Auditorium

4:05pm

4:30pm

High-Throughput 3D Cellular Models- Medium-scale Comparative Study of 2D vs. 3D Models Using High Content Imaging Approaches.
3D cellular models represent a fantastic opportunity for pharmaceutical industries to improve their discovery pipeline efficacy by bringing potentially much more relevant models within the early stages of the workflow. Moreover, it has been shown that in specific cases 2D and 3D models response to drug treatment can strongly vary and thus impact the in vitro relevance of the physiopathological model associated readouts. This approach is particularly well suited for complex or partially characterized targets in oncology. In this context, we have performed a systematic comparison of the 2D vs. 3D high content imaging readouts using a single cancer cell line. We will discuss about the challenges associated to such large scale screening in term of cellular processing but also in term of image and data analysis. Ultimately, we will discuss about the results generated from such a comparative study.

Speakers
avatar for Thierry Dorval, Ph.D.

Thierry Dorval, Ph.D.

Head of Data Science Lab, Servier
Thierry Dorval received a B.S. degree in theoretical physic and obtained a Ph.D. in image processing and artificial intelligence at Pierre & Marie Curie University, Paris, France. He then joined the Institut Pasteur Korea in 2005 first as researcher in biological image analysis then... Read More →

Chair

Monday October 21, 2019 4:30pm - 4:50pm
Wellcome Auditorium

4:50pm

High-Throughput 3D Cellular Models- Single-Cell Imaging and Advanced Image Analysis of Primary Tumors for Anticancer Therapy Development
The ability to perform high-content screening in a high-throughput fashion is routinely limited to cell lines and other explant model systems, however, there is a risk that these may not be fully representative of the in vivo environment due to culture adaptation or the lack of multi-lineage cell types. The ability to gather high-content data directly from primary samples however, both direct from blood and bone marrow or from metastasized cancers, without cell outgrowth or selection in a method amenable to laboratory automation can be a more direct system. Further, by combining imaging of these primary sample with an analysis pipelines robust to micro-aggregates, vastly different cell shapes and sizes, and that can ultimately harness the features from each cell can become a powerful means to study drug response in a variety of indications using model systems directly derived from the patient. This methodology has been used to prioritize therapy for late-state patients with hematological cancers in a basket trial (Snijder & Vladimer et al 2017,Lancet Hematology), has been integrated with genetic data to further uncover biological understanding and clinical synergy options (Schmidl & Vladimer et al 2019, Nat Chem Bio), and is now robustly being tested in technical validation studies for some indications to understand its potential use as an in vitro diagnostic.

Speakers
avatar for Gregory Vladimer, Ph.D.

Gregory Vladimer, Ph.D.

CSO, Allcyte
Gregory Vladimer is the CSO of Allcyte, a startup in Vienna, Austria. He received his PhD from UMass Medical School where he studied inflammation, and was a Senior Postdoctoral Fellow at CeMM in Vienna. Together with the Department of Hematology of the MedUniWien, spearheaded the... Read More →

Chair

Monday October 21, 2019 4:50pm - 5:10pm
Wellcome Auditorium

5:10pm

High-Throughput 3D Cellular Models-Patient Derived Scaffold (PDS) - A Platform for Optimised In Vivo Like Cancer Research and Clinical Testing
Cancer cells are surrounded and actively interact with the microenvironment at the primary site of growth as well as metastatic niches. Key components in the cancer environment have been linked to various aggressive cancer features and can further influence the essential subpopulation of cancer stem cells most likely governing malignant properties and treatment resistance. In order to model and specifically enumerate the influence of a specific cancer microenvironment we have developed a cell culture platform using cell free scaffolds from cancer samples infiltrated with standardized cancer cells. This in vivo like growth system induced a series of orchestrated changes in differentiation, EMT and proliferation of the cancer population with a final noticeable cancer stem cell expansion as defined by several surrogate assays and functional tests. The developed scaffold model system has a potential to optimally mimic in vivo like growth conditions reveling hidden and highly relevant clinical information about the malignancy inducing property of the specific scaffold earlier surrounding and indeed influencing cancer progressing properties. The platform can be utilized as a diagnostic tool, research instrument and in vivo mimic within screening program as will be further elaborated in the presentation.

Speakers
avatar for Goran Landberg, MD, Ph.D.

Goran Landberg, MD, Ph.D.

Professor, University of Gothenburg
Göran Landberg is professor and senior consultant in pathology focusing on molecular pathology and breast cancer. He has established several national and international centers focusing on cancer research both in Sweden and in England. In total, he has published more than 170 articles... Read More →

Chair

Monday October 21, 2019 5:10pm - 5:30pm
Wellcome Auditorium

5:30pm

High-Throughput 3D Cellular Models- Human 3D Neuronal Cultures for Pheontypic Drug Screening In Neurodegenerative Diseases
Several neurodegenerative diseases are characterized by axon degeneration. This is especially true for the peripheral neuropathies, a heterogeneous group of human diseases characterized by progressive degeneration of peripheral axons. Charcot-Marie-Tooth disease (CMT) is a group of peripheral neuropathies caused by a variety of genetic mutations leading to length-dependent axon loss. CMT is the most common neurogenetic disease, affecting 3 million people worldwide. It is a devastating, untreatable disorder. It limits patients’ ability to walk, balance themselves and use their hands and is associated with significant functional disability. It can also cause marked sensory impairment and neuropathic pain. Despite significant advances in our understanding of the pathophysiology of CMT, disease-modifying therapies are entirely lacking, partially due to lack of translational models suitable for drug discovery. One of the major challenges in creating such models is the need to specifically analyze axons in vitro. Two-dimensional neuronal cultures often exhibit overlap of cell bodies and neurites, making it difficult to analyze axonal morphology and potential screenable phenotypes in a high throughput manner. To overcome this limitation, we created a three-dimensional culture system based on induced pluripotent stem cell (iPSC)-derived motor neurons (spinal spheroids), one of the main cellular types affected by CMT. Spinal spheroids can be formed by culturing sorted iPSC-derived motor neurons in ultra-low-attachment 384-well plates. When transferred to laminin-coated plates, spinal spheroids attach to the bottom of the well, allowing for the robust growth of axons in a centrifugal fashion, optimal for high content imaging. Using this system, we identified a robust and reproducible axonal phenotype in spinal spheroids from patients with CMT2E, a particular type of CMT caused by missense mutations in NEFL (neurofilament light chain gene). This phenotype is characterized by the abnormal accumulation of neurofilaments in discrete areas of the motor axons and nicely recapitulates findings from a mouse model of this same disease. This phenotype is also easily quantifiable using automated image analysis. A focused, proof-of-concept experiment investigating the effect of kinase inhibitors on the CMT2E axonal phenotype identified two compounds capable of improving neurofilament distribution in motor axons, demonstrating the potential of this platform as a useful tool for drug discovery in CMT. High throughput drug screening of several compound libraries is expected to start soon.

Co-Authors:
Renata, Maciel, PhD, MBA
Banupriya, Sridharan, PhD
Louis, Scampavia, PhD
Timothy, Spicer, PhD

Speakers
avatar for Mario Saporta, MD, Ph.D., MBA

Mario Saporta, MD, Ph.D., MBA

Assistant Professor of Neurology and Human Genetics, University of Miami Miller School of Medicine
Mario Saporta, MD, Ph.D., MBA is an assistant professor of Neurology and Human Genetics at the University of Miami. He is a clinical neurologist and translational scientist specialized in neuromuscular genetic diseases. His lab focuses on the use of patient-derived cell lines to model... Read More →

Chair

Monday October 21, 2019 5:30pm - 5:50pm
Wellcome Auditorium

5:50pm

6:00pm

Networking Reception
Monday October 21, 2019 6:00pm - 7:00pm
Manby Gallery
 
Tuesday, October 22
 

8:00am

Morning Stroll Through the King's Cross Area
Departure and arrival will be from/at the Pullman Hotel.

Join for a walking tour around the historical St. Pancras and King Cross neighbourhoods where we will focus on the Victorian men and women who helped develop it and lived there throughout the process. The tour route will include visiting:

  • St. Pancras Renaissance Hotel – Victorian Gothic
  • Kings Cross Station
  • Granary Square: the canal/grain warehouse/hoists and equipment for moving goods
  • Handy side sheds, originally used for train, horse and cart transportation
  • Central St. Martin’s School of Art
  • Horse Stables
  • Coal Drops Yard
  • Gas Cylinders, history of the gas supply
  • German Gymnasium – Victorian British Library
  • British Library



Tuesday October 22, 2019 8:00am - 9:00am

8:00am

Registration Open
Tuesday October 22, 2019 8:00am - 2:00pm
Manby Gallery

9:00am

Vendor Workshop - MIMETAS and Molecular Devices - Developing High-Throughput Organ-On-A-Chip Tissue Models for Drug Discovery and Unlocking their Full Potential Using High-Content Imaging


Organ-on-a-Chip technology is a new paradigm in drug testing. The aim of this technology is to raise the physiological relevance of traditional cell culture by combining it with microfluidic techniques. Organs-on-a-Chip are 3D tissues that capture the complexity of in vivo tissues including 3D morphology, extracellular matrix embedment, multiple cell types, vascular structure and perfusion flow. Incorporating these technologies in a microplate format enables increased throughput in high content imaging systems.

In this presentation, experts from MIMETAS and Molecular Devices will introduce the culture and interrogation of complex 3D tissues, such as liver, kidney, gut and brain tissue. The tissues are grown in the OrganoPlate®, an in vitro cell culture microplate platform that allows culturing of over 40 tissues in parallel. The tissues are embedded in an extracellular matrix gel and comprise both stromal tissue, blood vessel structures as well as epithelial barriers. We will show examples of Organ-on-a-Chip tissues for use in disease modelling and drug safety testing and will present the new OrganoPlate® Graft, which allows vascularization of spheroids, organoids and PDX explants, and offers a powerful solution for the study of angiogenesis, an important focus for cancer therapeutics.

These complex 3D tissue cultures can be interrogated in a high-throughput manner using powerful cellular imaging systems. We will show how Molecular Device’s ImageXpress Micro Confocal High-Content Imaging System and ImageXpress Pico Automated Cell Imager play a vital role in the development and analysis of 3D tissue models, such as those built on the OrganoPlate® platform.

Tuesday October 22, 2019 9:00am - 10:30am

10:00am

Coffee in Exhibition
Tuesday October 22, 2019 10:00am - 10:30am
Manby Gallery

10:30am

Advances in Imaging and Analysis- Light Sheet Fluorescence Microscopy for High-Content Analysis in 3D
Light sheet fluorescence microscopy (LSFM) provides low out-of-plane photobleaching and phototoxicity, but usually requires two microscope objective lenses orientated at 90° to one another, making high content imaging in conventional well plates more challenging. Oblique plane microscopy (OPM) uses a single high numerical aperture microscope objective to provide both fluorescence excitation and detection whilst maintaining the advantages of LSFM. We present the development of a stage scanning OPM approach for light sheet fluorescence imaging in commercially available 96 and 384-well plates, including plastic-bottomed plates.
In a first application, the system was used to measure cell shape parameters in fixed and live cells in grown in 3D in a collagen matrix. We are developing a MATLAB 3D image analysis pipeline for automated segmentation and morphological quantification of the image data. This work aims to enable a better understanding of which genes are responsible for cancer cell size determination and invasion in 3D cultures.
In a second application, the system is being used to develop a 3D assay for spheroid-based compound safety screening. Multi-cellular HepG2/C3a spheroids with diameters in the range 100-200 micron were grown in low attachment u-bottomed dishes and then transferred to 384-well plates for ssOPM imaging. Following initial tests, SYTOX green and TMRM were chosen to read out spheroid viability and mitochondrial function as an indicator for drug toxicity. The assay Z’, signal to noise ratio and coefficient of variation have been measured using max-min plating protocols.

Speakers
avatar for Chris Dunsby

Chris Dunsby

Senior Lecturer, Imperial College London
Dr Chris Dunsby works on the development of quantitative fluorescence imaging techniques for applications in biomedicine, including Förster resonant energy transfer microscopy, automated multiwell plate imaging and fluorescence lifetime imaging (FLIM). He has also worked on super-resolution... Read More →

Chair

Tuesday October 22, 2019 10:30am - 11:50am
Wellcome Auditorium

10:50am

Advances in Imaging and Analysis- Imaging Rapid Cell Shape Transitions and Volume Control in 3D
Imaging rapid cell shape transitions and volume control in 3D
Regulation of cell shape in 3D space is essential during human development, and misregulation of cell shape is central to human disease states such as metastatic cancer. We are using 3D high content imaging to understand how healthy and diseased cells regulate their 3D shape and volume.

Approach: Oblique plane microscopy for 3D imaging
To make measurements in 3D we are taking advantage of recently developed Oblique Plane Microscopy (OPM). OPM imaging combines the benefits of light sheet microscopy - high speed 3D imaging, and low phototoxicity - with the advantages of conventional microscopy such as simple mounting techniques and high throughput sampling.

i) Cell shape transitions
To understand the molecular networks controlling 3D shape transitions in melanoma, we are live imaging cells embedded in collagen matrices. We are using this system to establish baseline shape dynamics, as well as shape transitions when cytoskeletal regulators are inhibited or when cells are in the presence of clinical drugs.

ii) Control of cell volume
Volume control is key to cell function because the volume of a cell influences the scale, duration and dynamics of all biochemical processes within. We have been using volumetric imaging by OPM in immortalised epithelial cells to understand how cell geometry is maintained throughout each stage of the cell cycle, and across cell generations. We anticipate these discoveries can form the basis of new strategies to target size control checkpoints in the treatment of cancers.

Speakers
avatar for Lucas Dent

Lucas Dent

Postdoctoral Fellow, Institute of Cancer Research
My research aims to understand the signalling networks controlling cell shape and adhesion during normal development, and how these are altered in disease. To do this, my approach is to combine Drosophila (fruit fly) genetics and mammalian (human) systems to reveal the broad principles... Read More →

Chair

Tuesday October 22, 2019 10:50am - 11:10am
Wellcome Auditorium

11:10am

Advances in Imaging and Analysis- Extracting Meaning From Big Data in Volume Electron Microscopy
Many different imaging modalities now routinely produce huge amounts of data thanks to increased acquisition speeds and extensive automation. Volume electron microscopy techniques, such as serial block face SEM (SBF SEM), focused ion beam SEM (FIB SEM) and array tomography (AT), produce datasets in the terabyte regime. Multimodal imaging methods such as correlative light and electron microscopy (CLEM) can be used to navigate the sample more efficiently, reducing the amount of data that must be analysed. We have developed a set of tools that allow us to "find the needle in the haystack" using these CLEM techniques.

Electron microscopy image data has remained stubbornly resistant to automatic computational analysis, with painstaking manual segmentation (finding and delineating a structure or object of interest) still being the gold standard. However, recent generations of microscope produce data far more quickly than a small team of experts can thoroughly analyse. Whilst new deep learning techniques offer significant promise in automating this analysis, the collection of sufficient annotations to provide training data is a major bottleneck. In collaboration with the Zooniverse (zooniverse.org) we have developed a citizen science project called “Etch a Cell” (etchacell.org) in which we ask volunteers to contribute segmentations. By collecting multiple segmentations per image, we are able to aggregate the volunteers’ annotations into accurate and robust data that can be used to train our machine learning system. Beyond the research applications, we have also found the project to be an effective outreach and education tool, letting students and non-experts gain an insight into the scientific process at the raw data level.

Speakers
avatar for Martin Jones, DPhil

Martin Jones, DPhil

Deputy Head of Microscopy Prototyping, The Francis Crick Institute
Martin Jones works in the Electron Microscopy Science Technology Platform at the Francis Crick Institute in London, developing new hardware and software for extracting meaning from ever more complex datasets.Martin's DPhil from Sussex University was in experimental atomic and quantum... Read More →

Chair

Tuesday October 22, 2019 11:10am - 11:30am
Wellcome Auditorium

11:30am

Advances in Imaging and Analysis- Development of a Multiparametric Structural Cardiovascular Toxicity Imaging Assay Using iPSc Derived Cardiomyocytes
According to Laverty and co-workers (2011), 27% of drugs fail to reach phase I due to cardiovascular liability and up to 45% of drug withdrawals post approval are due to cardiovascular toxicity. The majority of these withdrawals were due to the induction of arrythmia in patients which led to various initiatives by regulatory agencies such as the comprehensive in vitro proarrhythmia assay (CiPA) initiative by the FDA which utilise functional assessments of human iPSc derived cardiomyocytes to predict potential acute cardiovascular liabilities. However, these approaches cannot capture the effects of compounds with a chronic dosage regimen that are capable of causing structural changes in cardiac cells.

The aim of this project is to develop an in vitro assay capable of flagging compounds with potential cardiovascular liabilities when applied chronically using iPSc derived cardiomyocytes. In order to capture the multivariate potential mechanisms of toxicity, cells were measured to assess a number of different functional and structural parameters, such as calcium flux, contraction, viability, mitochondrial membrane potential as well as actin organisation. The use of 3D co-culture models was also assessed as these are postulated in the literature to increase the maturity and, therefore, the physiological relevance of the model.

The approach generated data from numerous parameters describing both kinetic and imaging readouts, requiring customised data analysis solutions. This brought the challenge to balance the need to test compounds rapidly in high throughput (384-well) at early lead identification and lead optimisation stages of drug development, with the desire to generate rich datasets informative of underlying mechanisms of structural cardiotoxicity.

Assay predictivity was assessed using annotated diverse compound sets including both proprietary and public domain compounds known to induce structural cardiotoxicity in vivo.

Authors:
Ellie Handford, GSK
Peter Clements, GSK
Andrew Brown, GSK
Jo Francis, GSK

Speakers
avatar for William Stebbeds, Ph.D.

William Stebbeds, Ph.D.

Senior Scientist, GSK
Senior scientist at GSK specializing in developing novel assays for drug discovery and development using complex in vitro models and multi-parametric analysis

Chair

Tuesday October 22, 2019 11:30am - 11:50am
Wellcome Auditorium

11:50am

11:55am

12:00pm

1:30pm

Poster Presentation #11-Using 3D Neural Spheroids to Accelerate Drug Discovery with Kinetic and High Content Imaging
Three-dimensional platforms of human induced pluripotent stem cell (hiPSC)-derived neurons enable greater physiological relevance and promise to breathe new life into drug discovery programs. A fundamental requirement of this re-vitalization is high-throughput and informative interrogation methodologies.StemoniX has developed microBrain®3D, ready to use spheroids comprised of human iPSC-derived cortical neurons and astrocytes supplied in ready to use 96- and 384-well plates.Real-time, high-throughput kinetic imaging has demonstrated the value of this preparation in functional screening and disease model settings while advancements in tissue clearing and spheroid processing are enabling in-depth, granular structural examinations. In particular, our data demonstrate a dramatic benefit of tissue clearing for confocal analysis of neuronal spheroids stained with antibodies and Cell Painting dyes as well as enhanced throughput enabled by physical stabilization of spheroids on flat-bottom imaging plates. This presentation will highlight recent uses of microBrain 3D in functional 384-well based screens, patient-derived disease models, phenotypic rescue, and image-based interrogation. Together 3D human neural spheroids with functional and structural experimental methodologies hold the potential to overcome several bottlenecks in CNS-based drug discovery.

Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #12-Feasibility of Manufacturing-scale Bioproduction of Novel Next-generation 3D Organoid Cancer Models in Support of the Human Cancer Models Initiative
To meet the need for improved approaches to study cancer in vitro, there has been a surge in the development of novel research models utilizing advanced culture methods. These methods permit in vitro growth of cancer types previously not possible, and/or models with enhanced in vivo relevancy compared to traditional continuous cell lines. However, availability of these early-stage research models is currently limited and there is a lack of data on the ability to scale up production of these models to support the needs of the global research community. We sought to investigate the protocols, expansion capacity, cryopreservation ability, genetic stability, and feasibility of larger-scale bioproduction of a subset of the models generated by the Human Cancer Models Initiative (HCMI), an international collaborative effort between Cancer Research UK, the foundation Hubrecht Organoid Technology, the National Cancer Institute, and the Wellcome Sanger Institute. The HCMI’s initial goal is the development of 1,000 novel human cancer models, paired with bioinformatics and patient clinical data, particularly from rare or underrepresented cancer types. One advanced culture method being utilized, three-dimensional organoid “microtissue” culture, potentially poses challenges for traditional large-scale bioproduction processes. It requires growth embedded within an undefined extracellular matrix and complex media formulations containing multiple small molecules and recombinant proteins with unknown stability and shelf-life. Additionally, organoid growth media typically includes multiple sources of undefined conditioned media containing critical growth factors. We cultured organoid models derived from human colon, pancreas, esophagus, and mammary tissues developed by laboratories contributing to the HCMI. Multiple unique donors were available for all tissues and both cancer and non-cancer models were available for two tissue types. Most models were maintained in culture continuously for at least 60 days (7-27 population doublings, > 10 passages). Tissue and donor variability was evident in model characteristics, including morphology (assessed by microscopy and immunocytochemistry), growth rate, and genetic stability (measured by short tandem repeats analysis). All models were amenable to scale up beyond multiwell plates, and all models could recover from cryopreservation. While organoid culture represents a significant divergence from typical two-dimensional monolayer culture of continuous cell lines, our results show that these next-generation in vitro models are suitable for larger-scale bioproduction. This is vital to ensure the widespread availability of these models within the research community to facilitate applications like pre-clinical drug discovery and basic cancer research.

Speakers
NP

Nirmal Perera, Bachelor of Medicine, Bachelor of Surgery (M.B.B.S), Medicine. Masters of Research, Genetics and Gene The

ATCC Field Applications Scientist, LGC
Dr. Nirmal Perera is a Technical Sales and Field Application Specialist for ATCC with LGC. He's been in his current role for almost 2 years now, attending and presenting at conferences all over the world.


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #13-A Functional Study of the Endomembrane System in Mammalian Cells Growing in a 3D Environment
The human body is a complex assembly of tissues and organs, each of which is made of different types of cells. Specialisation of function is achieved via compartmentalisation within each cell, as well as defined communication networks that exist between endomembrane compartments. The endomembrane system is essential for the synthesis and distribution of proteins, lipids and carbohydrates, and as such needs to be tightly regulated. One protein family of master regulators of the endomembrane system is the Rab small GTPases. Rab proteins govern the communication between organelles, via the GDP-GTP cycle and by exhibiting specific membrane localisation patterns. To date, their distribution and function have mostly been investigated in conventional two-dimensional in vitro cell models, however three-dimensional cell models, so far applied to investigate cancer and drug development, offer new possibilities. In this work, a functional study is being used to understand the localisation and role of Rab proteins in the endomembrane system in a three-dimensional environment. For this, a three-dimensional cell model (spheroids), using HeLa Kyoto cells grown on micropatterned plates, has been developed. This approach allows highly uniform spheroids to be grown in a tightly controlled manner. In order to localise various Rab proteins, stably expressing cell lines expressing GFP-tagged Rab proteins have been produced and grown as spheroids. Automated high-content screening microscopy is now being used to quantitatively describe the localisation and function of Rab proteins in this new cell system, and in turn, compare the results to existing data coming from classical two-dimensional cell models.

Speakers
MM

Margaritha Mysior, B.Sc., M.Sc.

PhD candidate, University College Dublin
I absolved a Bachelor (2012-2015) and Master (2015-2017) degree program at the University of Applied Science Gelsenkirchen, Recklinghausen and Bocholt. During my Master thesis (October 2016 - August 2017) I characterised five unidentified regulators of the retrograde transport pathway... Read More →


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #14- GBM Infiltration of Cerebral Organoids Models Brain Tumour Invasion
Introduction
Glioblastoma (GBM) is the most common and most aggressive brain tumours in adults. No cure is available and complete surgical resection is not achievable due to GBM cell infiltration into the healthy brain parenchyma. Therefore, interventions that specifically target the invasive GBM phenotype are highly desirable. However, it is challenging to model the process of GBM infiltration in real time in the laboratory in relevant model systems encompassing both tumour and neural tissue compartments. Previously we demonstrated the ability of patient derived GBM spheroids to spontaneously infiltrate mouse derived early-stage cerebral organoids (CO). Here we present an enhanced culture system enabling tractable ex vivo investigation of human GBM into human-derived CO.

Methods
Adaptation of previously described methods in murine CO has allowed us to see spontaneous infiltration of GFP-expressing GBM patient derived cell model spheroids into early stage human COs and quantified using live cell imaging and histological analysis.

Results
Patient derived GBM infiltrated in all cases CO whereas neural progenitors (NP) were unable to infiltrate. The GBM spheroid and CO self-assembly and tumour cell infiltration were characterised quantitatively using several parameters including total GBM cell infiltration, migration distance, and cell compartment spread in 3D. This ‘assembloid’ assay was also able to differentiate ‘early’ and ‘late’ invasion capabilities of GBM cell models from different patients and GBM subtypes, as well as successful infiltration of a freshly resected tumour piece.

Conclusion
We demonstrate that GBM/CO assembloids can be adapted for scalable imaging applications (for a throughput of n>100 and >10 conditions per time lapse experiment). They can be used as an ex vivo experimental model system that aims, in real-time, to recapitulate the infiltrative characteristic of GBM that makes current therapy so ineffective.

Looking forward, in collaboration with the Stem Cell Hotel (King’s College London) and industry partner Cellesce (Cardiff), we plan to automate the quantification of multiple relevant imaging parameters that may enable predictive biological readouts in the future. Furthermore, we plan to use brain tumour assembloids to establish proof-of-principle work for laboratory testing of personalised treatments as a clinical decision system that may aid treatment stratification of brain tumour patients.

Speakers
BI

Bronwyn Irving, MSci

PhD Student, University of Leeds
Bronwyn is currently a PhD student at the University of Leeds following her passion for cancer research after previously working at UCL’s Institute of Child Health and Newcastle University’s Northern Institute for Cancer Research.


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #15- High Content Screening Platforms for the Evaluation of Steatosis Induction in 2D and 3D HepG2 cultures
Steatosis, or the increased accumulation of lipids in the liver, can be induced by several factors including exposure to steatogenic drugs and through pertubations in lipid metabolism indicative of non-alcoholic fatty liver disease (NAFLD). Hepatic lipid accumulation is a key driver of morphological and functional changes to liver architecture that leads to the development of acute and chronic liver disease and fibrosis. We have developed a number of high content screening platforms to screen for drug induced steatosis using 2D HepG2 cells and 3D HepG2 C3A spheroid cultures with detection of lipid accumulation by LipidTOX green and Nile red.Additionally, we have adapted our 3D spheroid platform to evaluate steatosis induced by dietary fatty acid uptake. We demonstrate our high throughput screening platforms utilising automated ECHO650 and Biomek NxP compound dispense combined with High Content Imaging of lipid accumulation using the ImageXpress Micro Confocal. Our assays are highly reproducible and have been validated using a panel of known steatogenic drugs. Development of such platforms will enable the early screening of candidates with steatogenic potential with a view to de-risking and de-selecting adverse compounds during the Discovery phase of drug development.

Speakers
KO

Katalin Orban, MSc, PharmD

Senior Scientist, Sygnature Discovery
Katalin Orban currently works in the DMPK and Physical Sciences Department at Sygnature Discovery where she contributes to the strategy, interpretation and overall DMPK assessment of NCEs in small molecule drug discovery.Katalin previously graduated from the Semmelweis University... Read More →


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #16- Genedata Imagence®: An Evaluation of Deep Learning for High Content Analysis
Machine learning has seen some revolutionary and remarkable developments over the last few years. Exciting breakthroughs have been achieved in artificial neural networks, in particular around deep network architectures (= deep learning), which work based on raw-pixel image information and provide a classification accuracy exceeding human expert judgement. Deep neural networks are now the state-of-the-art machine learning models across a variety of areas, including image processing and analysis and are widely implemented in academia and industry.

Genedata has developed the first commercially available solution for deep learning-based High Content Image analysis. Genedata Imagence® allows for the application of deep networks to the analysis of High Content Imaging, creating a workflow that cuts image analysis time, increases data quality, reproducibility of results and seamlessly integrates with Genedata Screener® for image data analysis. This deep learning approach outperforms conventional approaches for feature extraction and phenotype classification.

Here we provide the evaluation of a recent pilot of Genedata Imagence® for High Content Analysis. We compare the Genedata Imagence® workflow with our conventional in-house High Content Imaging and Analysis workflow through a series of assays that have been developed within the target validation biology group at LifeArc. These include, immunocytochemistry of shRNA knock-down clones, antibody internalisation assay, High Content cell health assay, growth cone collapse assay and osteoclast differentiation assay.The generation of training data and subsequent training of the network was carried out for each assay and results of the data analysis were compared to in-house conventional analysis. We discuss a few limitations but show overall an excellent data quality produced by this novel deep learning module for High Content Image analysis.

Speakers
ZI

Zaynab Isseljee

Scientist, LifeArc
Zaynab Isseljee is an enthusiastic research scientist with ten years’ experience in drug discovery. Graduated with a BSC (Hons) in Biomedical Science at King's College London with an industrial placement year at Novartis Horsham Research Centre. Since 2009, Zaynab has worked at... Read More →


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #17- Deep Learning-Derived Features Outperform Classical Computer Vision in Low Dimension-Embedding of High-Content Screening Data
When automating analysis of high-content screening images, a key step is reducing the object image (e. g. a cell) to a vector of features that describe that object for subsequent comparison with other objects. In classical computer vision, many of these features have an implicit biological meaning (e. g. cell size, intensity, aspect ratio, etc).

Depending on the research question, feature vectors then are subjected to univariate or multivariate analysis. Nonlinear dimensionality reduction techniques such as tSNE and its variants [1] are particularly useful when representing or embedding high-dimensional data in two or three dimensions.Ideally, such visualizations should clearly separate different phenotypes observed in the experiment. However, the embedding quality relies heavily on the quality of the input features derived from the HCS images.

In this poster, we use a high-content screening translocation assay to compare the quality of embedding produced by features extracted using classical versus deep learning-based feature approaches. We compare them based on ability to separate phenotypes and robustness against batch effects. We show that while classical and deep-learning-derived feature sets or a combination of both all produce excellent results, when using classical features, attaining this level of quality requires expert tuning of the feature extraction process to the assay of interest. In contrast, deep-learning-based feature extraction is fully automated and does not require expert knowledge.

Using deep learning-based features, we observe that the embedding quality depends on the network architecture. Standard CNN architectures perform poorly, while tailored architectures outperform classical methods. Finally, we show that for embedding and visualization, adding classical features to the deep-learning based features does not increase resolution and is thus unnecessary.

[1] L. J. P. van der Maaten and G. E. Hinton. Visualizing High-Dimensional Data Using t-SNE. Journal of Machine Learning Research 9(Nov):2579-2605, 2008

Speakers
MF

Matthias Fassler

Scientific Account Management, Genedata
Matthias Fassler is a Scientific Account Manager at Genedata, Basel.


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #18- Novel Organ-on-Chip Microfluidic Device based on Hydraulically Actuated Hydrogel Layers
Organs-on-chips (ooc) are small devices aiming to recapitulate the physiology and/or pathology of a living organism. Contrarily to commonly used flasks and dishes, ooc devices are designed to recapitulate the physico-chemical microenvironment of the cells. Thanks to these mimics of higher relevance, ooc devices are able to enhance cell differentiation compared to 2D culture techniques.
The field of ooc is still young, and many improvements remain to be done. For instance, one of the first organ-on-chip, produced by Huh et al. (2010), submitted epithelial and endothelial cells to mechanical stimuli by stretching a thin perforated PDMS membrane on which cells were attached. While extremely efficient, this setup presents several shortcomings, such as the high stiffness of PDMS compared to that of soft living tissues, and the poor relevance of a perforated membrane to study extravasation.
Here we present a novel ooc microfluidic device allowing the coculture of cells in and on a soft hydrogel. Thus, a greater tissue complexity can be mimicked, including notably a “stromal compartment” in addition to the usual epithelium-endothelium dual culture. Relying on a pressure differential between each side of the hydrogel and resistances to adjust the flow, the hydrogel can be stretched. Thus, in the final setup, most physico-chemical parameters (stiffness, flow, strain, functionalization, etc.) are under the control of the operator to best suit the desired organ and application.

Nicolas MINIER1, 2, Samy GOBAA1

Speakers
NM

Nicolas Minier

Institut Pasteur


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #19- Fully Automated and Scalable 3D-Baed Toxicity Testing in Human Neural Microtissues

Recent advances in stem cell technology have led to the development of three-dimensional (3D) culture systems, which have fueled hopes to bring about the next generation of more physiologically relevant high throughput screens (HTS). However, current protocols yield either complex but highly heterogeneous aggregates (“organoids”) or 3D structures with less physiological relevance (“spheroids”). Here, we present a fully scalable, HTS-compatible workflow for the automated generation, maintenance, and high content analysis of human midbrain-like automated microtissues (mANTs) in a standard SBS 96-well format. The resulting microtissues possess a highly homogeneous morphology, size, global gene expression, cellular composition, and internal organization. They present significant features of the developing human midbrain and display spontaneous aggregate-wide synchronized neural activity. By automating the entire workflow from generation to analysis, we provide essential intra- and inter-batch reproducibility as demonstrated via RNA sequencing and quantitative whole mount high content imaging. Dose response toxicity testing showcases our ability to quantify drug effects on single cells within a complex 3D environment using mANTs. In conclusion, our workflow has the potential to form the basis for 3D-tissue-based screening in a variety of applications including toxicology studies, disease modelling, and drug development.
 

Speakers
HR

Henrik Renner, Msc.

PhD Student, Max Planck Institute for molecular Biomedicine
- PhD student in the lab of Hans Schöler at the MPI in Münster (since 2016), Germany (creating 3D HTS tissue models for midbrain)- Msc. Karolinska Institute (2016)- Bsc. University of Göttingen (2014)


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #20-AI-driven Optimization of Multidrug Combinations in Heterotypic 3D Human Co-cultures
Heterotypic three-dimensional (3D) short-term human cellular models were developed based on the histopatological patient tissues to predict the efficacy of new (combined) treatment regimens for treatment of various stages of human colorectal carcinoma (CRC). These spheroidal callular co-cultures included human CRC cell lines of various genetics and stage co-cultured with human endothelial cells and fibroblasts.

The exposure of those spheroids with several low-dose three- or four-drug combinations, designed using artificial-intelligence-based method allowed efficient optimization of the most effective treatment strategy.

The optimized synergistic cell type-specific drug combinations induced cell metabolic activity inhibition, that indirectly corresponding to cell viability inhibition, by up to 90%. Moreover, the activity of those optimized drug combinations was often more potent in 3D models as compared to simple 2D cell cultures and facilitated further translation to more complex in vivo model.

Summarizing, our technology represents an innovative step to obtain enhanced accuracy of the activity of cell type-specific treatments. Our multicellular spheroids resemble in many aspects solid tumors and serve as a promising model to mimic the in vivo situation.

Speakers
PN

Patrycja NowakSliwinska, Ph.D., D.Sc.

Professor, Institute of Pharmaceutical Sciences of Western Switzerland
She performed her post-doctoral work in the Medical Photonics Group at the Swiss Federal Institute of Technology (EPFL) in Lausanne, Switzerland, and in the Angiogenesis Laboratory at the VU University Medical Center in Amsterdam, The Netherlands. She obtained the certificate of advanced... Read More →


Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

1:30pm

Poster Presentation #21- Icy2.0: An open and free software for bioimage analysis
We present Icy2.0 (http://icy.bioimageanalysis.org), which is a major evolution of our previous free and open software for bioimage analysis Icy. This release has three important features: 1) the ability to handle very large images (> 4Gb); 2) the possibility to process long sequences through an optimized streaming procedure with adaptive caching; and 3) a totally redesigned web site for improved interactions within the Icy community. Icy2.0 has retained and consolidated the major features that made its success in the bioimaging community, namely to make it available to the bioimaging community an image analysis software suite that encompasses the large variety of biological applications (microscopy, particle tracking, HCS/HTS, digital pathology, animal behavior…) and users (biologist, bioimage analyst, physicist, developer) and to give access to advanced image analysis methods and solutions.

Since its launching, it has been the constant philosophy of the Icy team to promote sharing of source code and know-how and to facilitate the use of quantitative approaches and open new scientific perspectives in terms of exhaustiveness, reproducibility and robustness of the analysis of bioimaging data sets. For those reasons, Icy2.0 has a fully redesigned web-site, that includes new communication channels between the end-users and the developers, new tutorial material and improved maintenance cycles. Icy2.0 now provides more than 400 dedicated plugins covering a large variety of state-of-the-art image analysis methods ranging from active contours models to Machine Learning through statistical spatial analysis, which empower users with the most recent and adapted quantitative image analysis and visualization tools. Protocols, which are a graphical front-end that enable software development without any programming knowledge, have also been improved and now include the possibility to develop sophisticated image processing pipelines in a more readable and interactive manner.

Icy2.0 has a large community of developers and users (4000+ regular users, 700+ students trained, 1500+ visits per month) that are using Icy for distributing their algorithms and plugins. For its long-term sustainability and development, the Icy2.0 project benefits from the institutional support of the French national Infrastructure FranceBioImaging and of Institut Pasteur. Ongoing work aims at improving the interoperability and convergence of Icy2.0 with other open and free image analysis packages as well as facilitating the integration and use of AI packages.



Speakers

Tuesday October 22, 2019 1:30pm - 2:00pm
Sherry Coutu Seminar Suite Foyer

2:10pm

Complex Translational Models-Development of Advanced Models of Haematopoiesis and their Application to Cancer Therapeutics
Haematological toxicity is a common safety challenge for many monotherapy and combination cancer therapies which can result in dose reductions and discontinuation to the point where the drugs become ineffective. To address the challenge, we are developing advanced in vitro models using primary human CD34+ hematopoietic stem and progenitor cells (HSPC’s) to assess drug induced haematological toxicity. Our aim is to provide human relevant in vitro data to inform on monotherapy and combination bone marrow risk and to optimise rational drug combination choices and scheduling in the clinic. Recent advances in 2D and 3D microphysiological systems (organ-on-a-chip) have opened new possibilities to investigate different aspects of haematological toxicity. Using CD-marker profiling we have defined the differentiation kinetics of HSPCs to either erythroid, myeloid, or megakaryocytic lineages. 2D systems allow a high throughput assessment of monotherapy or combinations risk whilst 3D system with continuous stem cell re-population and assay longevity also allows the capability to assess recovery and dose scheduling. The initial validation of these systems is very encouraging and appears to closely resemble the lineage specific hematological toxicity observed clinically.

Speakers
avatar for Mark Anderton, Ph.D.

Mark Anderton, Ph.D.

Director of Drug Safety, AstraZeneca
Mark Anderton is a Director of drug safety within the Oncology Safety department of AstraZeneca. He earned his PhD in Cancer Pharmacology and Toxicology at the MRC Toxicology Unit, Leicester. After his PhD, Mark joined Vertex Pharmaceuticals, UK where he gained experience in all... Read More →

Chair
TS

Timothy Spicer, BS, MS, Ph.D.

Senior Scientific Director, Scripps Research Florida


Tuesday October 22, 2019 2:10pm - 2:30pm
Wellcome Auditorium

2:30pm

Complex Translational Models- Building a Multicellular 3D Model of the Human Salivary Gland to Study Immune Mechanism Underlying Sjogren’s Syndrome
Sjogren's Syndrome (SS) is a chronic autoimmune condition causing exocrine tissue dysfunction, mainly in women, with a prevalence of around 1-3% in the general population. Most notably the lacrimal and salivary glands (SGs) are affected resulting in common symptoms of dry eyes and dry mouth, the latter associated with increased dental caries and fungal infections. Other symptoms may include dry skin, joint pain, fatigue, and swollen SGs. Currently there are no effective therapies for this condition and treatment is limited to palliative management of the symptoms.

At the cellular level, SS is characterised by lymphocytic infiltration in the lacrimal and SGs, as well as other exocrine glands in the body, resulting in tissue destruction. This adaptive immune response can manifest as lymphocytic foci in the tissue that develops and functions as ectopic germinal centre like structures. Activation of innate immune pathways may play an early role in SS and precede lymphocytic infiltration. In the SGs, dendritic cells (DCs), macrophages, salivary gland epithelial cells (SGECs) and natural killer cells (NK) are thought to be involved.

Animal models such as mouse and non-human primate are often used to study SS and to assess molecular targets for novel therapeutics. However, these have their limitations in that the chronic disease state is difficult to reproduce, while no single animal model exhibits all of the clinical characteristics associated with the condition. Progress has been made in developing SG in vitro models to study SS. These have employed animal and human cell lines, as well as primary SGECs derived from SG tissue of healthy subjects and SS patients. These have been cultured on plastic or permeable supports, while some have attempted co-culturing with immune cells and others have shown 3-Dimensional tissue structures in collagen/matrigel.

In order to build an organotypic model of the human salivary gland it is important to bring together appropriate cell types in a microenvironment where there is adequate interplay between these cells to form a 3-D architecture that closely resembles the tissue in vivo, as well as anatomical and functional features of the individual cell types. A collagen-fibroblast matrix component (stromal matrix) has been shown to promote SG acinar cell function, therefore including this element into an in vitro model is a sensible option. It has also been shown that DCs exist in substantial numbers in the acini, ducts and interstitial areas of the SG epithelium, as well as other immune cells.
Our work focussed on establishing a normal multicellular 3-D human SG model, by co-culturing various immune cell types with a collagen-fibroblast matrix and a SG epithelial layer in an air lifted microenvironment. We encountered optimisation challenges for keeping all cell types viable in this model up to 12 days on a robust platform, suitable for various applications of drug discovery and translational research. We have characterised the morphological and functional features of the different cell types to ascertain the potential for generating a SS like disease model.

Co-Authors:
1 Martin Vidgeon-Hart; GSK
2 Paul McGill; GSK
3 Jan Klapwijk; GSK
4 Emma Koppe; GSK
5 Timothy Radstake; Utrecht

Speakers
avatar for Anita Naidoo, MPhil, BSC

Anita Naidoo, MPhil, BSC

Scientific Leader, GlaxoSmithKline Research and Development
Anita joined GlaxoSmithKline as a Genetic Toxicologist in 1988 and then moved into Cellular Pathology & Toxicology and re-trained as an in vitro toxicologist, applying her knowledge and expertise to develop cellular models to address safety concerns through bespoke mechanistic studies... Read More →

Chair
TS

Timothy Spicer, BS, MS, Ph.D.

Senior Scientific Director, Scripps Research Florida


Tuesday October 22, 2019 2:30pm - 2:50pm
Wellcome Auditorium

2:50pm

Complex Translational Models- Modelling the Human Airway Mucosa for Preclinical Drug Development and Testing
In the UK, lung diseases account for 20% of deaths, >700,000 hospital admissions and >6 million inpatient bed-days/year. Admissions often result from acute exacerbations for which there are no effective treatments, despite large investment by pharma. Over 85% of promising new drug candidates fail in clinical trials leading to high rates of attrition. Likely explanations include the poor correlation between current animal models with the human pathology and the lack of representative, validated and qualified 3D human cell models. In our clinical translational studies, we use human lung tissue samples to study disease mechanisms and are developing the ‘4D Airway Biochip’ platform to help improve our understanding of lung diseases.  This microfluidic platform models interstitial flow, providing a cellular environment that is closer to the in vivo situation, and allows kinetic sampling of cellular secretions.  Furthermore, it provides an air interface for challenge with environmental agents,  whilst frequency-dependent electrical impedance measurements allow epithelial barrier properties to be monitored in real time.  Data from this platform suggest that it is predictive of in vivo tissue responses and offers a practical solution for new drug discovery not only in airway diseases but for other diseases where epithelial barrier defects contribute to disease pathogenesis. It may also offer a useful in vitro platform for pharmacological and toxicological studies.

Speakers
avatar for Donna Davies, Ph.D.

Donna Davies, Ph.D.

Professor of Respiratory Cell and Molecular Biology, Clinical and Experimental Sciences, University of Southampton
Donna Davies has a PhD in Biochemistry and holds a Personal Chair at the University of Southampton, UK. She has pioneered the use of tissue engineering using airway tissue derived from volunteers with asthma or COPD as alternatives to using animal models of pulmonary disease.  More... Read More →

Chair
TS

Timothy Spicer, BS, MS, Ph.D.

Senior Scientific Director, Scripps Research Florida


Tuesday October 22, 2019 2:50pm - 3:10pm
Wellcome Auditorium

3:10pm

Complex Translational Models- Metastatic Colorectal Cancer Organoids as a Novel Model System for Personalized Medicine and Reverse Translation
Regorafenib has shown anti-cancer activity in metastatic colorectal cancer (mCRC) patients by inhibiting tumor vasculature. A mCRC PDO model can mimic tumor-stroma interaction for improving patient selection and measure drug resistance. In this work, we recapitulated clinical response in vivo, and compared to the ex vivo tumor-stroma model. A translational phase II trial of regorafenib in chemo-refractory mCRC patients with biopsiable metastases was conducted. For the in vivo models, PDOs were implanted in livers of NSG mice and treated with regorafenib. The Ex vivo model was generated by co-culturing PDOs with CAFs and EC. PDOs retained genomic and transcriptomic features of parental biopsies. The results showed vascular density reduction after regorafenib treatment in mice from responders’ patients, and no significant changes in non-responders. The developed PDO co-cultures resembled the metastatic niche predicting response to anti-cancer treatments to inform clinical decisions.

Co-Authors:
1 Georgios Vlachogiannis
2 Andrea Lampis
3 Khurum Khan
4 David Cunningham
5 Matteo Fassan
6 Ruwaida Begum
7 Leo Chan
8 Ning Lai
9 Reem Eldawud
10 Nicola Valeri

Speakers
avatar for Somaieh Hedayat

Somaieh Hedayat

Ph.D. student, Institute of Cancer research
Patient-derived organoids (PDOs) have recently emerged as robust preclinical models. We have recently shown that PDOs from metastatic, heavily pretreated, colorectal and gastroesophageal cancer patients mirror the phenotype and the genotype of their parental biopses and recapitulate... Read More →

Chair
TS

Timothy Spicer, BS, MS, Ph.D.

Senior Scientific Director, Scripps Research Florida


Tuesday October 22, 2019 3:10pm - 3:30pm
Wellcome Auditorium

3:30pm