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Plenary Session [clear filter]
Monday, October 21
 

11:30am BST

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 BST
Wellcome Auditorium

11:50am BST

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 BST
Wellcome Auditorium

12:10pm BST

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 BST
Wellcome Auditorium

12:30pm BST

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 BST
Wellcome Auditorium

3:00pm BST

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 BST
Wellcome Auditorium

3:15pm BST

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 BST
Wellcome Auditorium

3:25pm BST

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 BST
Wellcome Auditorium

3:35pm BST

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 BST
Wellcome Auditorium

3:45pm BST

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 BST
Wellcome Auditorium

3:55pm BST

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 BST
Wellcome Auditorium

4:30pm BST

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 BST
Wellcome Auditorium

4:50pm BST

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 BST
Wellcome Auditorium

5:10pm BST

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
GL

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 BST
Wellcome Auditorium

5:30pm BST

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 BST
Wellcome Auditorium
 
Tuesday, October 22
 

10:30am BST

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 BST
Wellcome Auditorium

10:50am BST

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 BST
Wellcome Auditorium

11:10am BST

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 BST
Wellcome Auditorium

11:30am BST

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 BST
Wellcome Auditorium

2:10pm BST

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 BST
Wellcome Auditorium

2:30pm BST

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 BST
Wellcome Auditorium

2:50pm BST

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 BST
Wellcome Auditorium

3:10pm BST

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 BST
Wellcome Auditorium