Monday, 13 November 2023

08:00

Conference Registration and Materials Pick-Up + Coffee in the Exhibit Hall

09:00

Dominique PV de KleijnConference Chair

Introduction to the EV Field -- circa 2023
Dominique PV de Kleijn, Professor Experimental Vascular Surgery, Professor Netherlands Heart Institute, University Medical Center Utrecht, The Netherlands, Netherlands

09:30

Alain BrissonKeynote Presentation

Contribution of Immuno-Cryo-Electron Microscopy to the EV Field
Alain Brisson, Professor Emeritus, University of Bordeaux, France

Extracellular vesicle (EV) samples isolated from body fluids or conditioned media are heterogeneous in cell origin, membrane compartment origin, size, molecular composition and functional properties. Given the diversity of biological functions and biomedical applications borne by EV, there is an urgent need for efficient and reliable methods of EV characterization and isolation, to improve our understanding on EV and fully exploit their potential. Our group has pioneered the use of cryo-transmission electron microscopy (cryo-EM), immuno-gold labeling and flow cytometry for imaging and quantifying EV. In this presentation, I will first summarize the main contribution of immuno-cryo-EM to the EV field, helping to answer the question “What do contain EV samples?”. Next, I will present recent results and discuss our attempts to identify EV-based biomarkers in sickle cell disease and preeclampsia.

10:00

LONZACharacterization of Cell-Derived Vesicles: Are They Really Different from Small EVs?
Ilaria Passalacqua, Scientist, Process Development, LONZA

Cell-derived vesicles (CDVs) are one of the most promising non-viral drug delivery platforms. High amounts of these vesicles are produced through mechanical extrusion of cells and used to carry payloads with therapeutic benefit. CDVs share molecular signatures found in the producer cells but their cargo may differ from other classes of extracellular vesicles (EVs) due to their peculiar genesis. In this study, we ought to identify biophysical and molecular characteristics unique to CDVs or shared with small-EVs. Lonza’s HEK293 cells were used to produce CDVs using MDimune’s proprietary extrusion technology and to purify small-EVs from conditioned media. Both EV types were purified using the same downstream process and characterized by mass spectrometry and nano-flow cytometry (nFCM) to identify CDV-specific markers and evaluate membrane integrity. CDV-enriched proteins were found by proteomic analysis and confirmed by nFCM. Specifically, LAMP-1 and CD63 were identified as CDV-enriched markers and further confirmed by western blot analysis and flow cytometry (FACS). Notably, a partial overlap was observed between CD63 and LAMP-1 positive EV subpopulations , suggesting distinct molecular properties and, perhaps, functional roles. Further work is warranted to confirm these observations and to better understand the potential of CDVs a non-viral vector modality for cell-and-gene therapies.

10:30

Mid-Morning Coffee and Tea Break and Networking in the Exhibit Hall

11:15

Samir EL-AndaloussiKeynote Presentation

Gene Editing and Targeted Delivery Using Engineered Extracellular Vesicles
Samir EL-Andaloussi, Associate Professor, Karolinska Institutet, Sweden

Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication due to their ability to transfer bioactive lipids, proteins and different species of RNA into cells. As such, EVs can be harnessed for the delivery of macromolecular drugs. Adapting EVs for drug delivery requires cellular engineering that allows for selective loading of biotherapeutics inside or on the surface of EVs. This presentation will cover our recent developments in EV engineering, with focus on active loading and functional delivery of gene editing modalities. In addition to addressing PK, PD and functional delivery of EVs in normal and diseased mice, strategies to enhance tissue targeting and extending plasma half-life of circulating EVs will be discussed.

11:45

NanoFCM Co., Ltd.NanoFCM Brings Flow Cytometry Capabilities to the Nanoscale!
Natalia Gebara, Application Scientist, NanoFCM Co., Ltd.

Conventional flow cytometers often struggle to meet the sensitivity requirements for the analysis of nanoscale particles, such as exosomes, nanomedicine, and viruses. To meet this challenge, NanoFCM has developed the NanoAnalyzer, a dedicated nano-flow cytometry platform, which offers a flexible and high-throughput solution for sub-micron analysis. By using the NanoAnalyzer, single-particle characterization can be achieved which simultaneously measures the side scatter (40 -1000nm) and fluorescent properties of particles. The size detection of the NanoAnalyzer favorably compares to electron microscopy and covers the entire size range of EVs, offering a detailed analysis of size, concentration, and biochemical properties by direct correlation of the physical and phenotypic data.  It is by combination of all these properties that the NanoAnalyzer is an ideal next-generation technique/ instrument for the analysis of EVs.

12:15

Networking Buffet Lunch in the Exhibit Hall -- Networking with Colleagues, Engage with Exhibitors


Session Title: Technologies and Tools for Studying Extracellular Vesicles (EVs)

13:30

Cytek BioSciences Technology Spotlight Presentation

14:00

Particle Metrix GmbH and CEO, Particle Metrix Inc., USAColocalization of Biomarkers with the New Particle Metrix ZetaView x30 Family
Sven Rudolf Kreutel, Chief Executive Officer, Particle Metrix GmbH and CEO, Particle Metrix Inc., USA

During the last decades, Nanoparticle Tracking Analysis (NTA) has emerged as a vital and fast characterization technology for biological nanoparticles like Extracellular Vesicles (EVs), Exosomes and Viruses. While classic NTA scatter operation feeds back particle size and total concentration, the fluorescence detection capability (f-NTA) enables the user to gain specific biochemical information. Statistical determination of signal colocalization on low nano scale particles however, is a challenging task for any analytical instrument including flow cytometers and microscopes. A new laser generation paired with ultra fast switching times between fluorescence channels lays the foundation of colocalization nanoparticle tracking analysis (C-NTA) introduced recently by Particle Metrix. For the first time, we report results of colocalization measurements on reference material as well as real biological nanoparticles based on NTA technology on the new ZetaView® PMX-430 QUATT.

14:30

SpectradyneAccurate Nanoparticle Size, Concentration, and Payload with Spectradyne’s ARC Particle Analyzer
Jean-Luc Fraikin, CEO, Spectradyne

Spectradyne’s ARC particle analyzer uses a unique combination of electrical and optical measurement techniques to accurately measure the size, concentration, and internal and external payload of nanoparticles as small as 50 nm in diameter.  Learn how scientists are using the ARC to quantify single-particle encapsulation efficiency for LNPs and characterize subpopulations of extracellular vesicles based on surface marker expression profiles.

15:00

Jiong-Wei WangKeynote Presentation

Targeting Hepatocytes with the Lipid Nanoparticular System in Fatty Liver Disease
Jiong-Wei Wang, Assistant Professor, National University of Singapore, Singapore

Non-alcoholic fatty liver disease (NAFLAD) is a spectrum of chronic liver disease caused by excessive fat accumulation in the liver, with a prevalence of up to 40% in the United States and in Singapore. NAFLAD can develop into a more severe form, non-alcoholic steatohepatitis (NASH), characterized with liver inflammation and fibrosis, and ultimately cirrhosis and liver cancer. Currently, efficacious drugs reversing the various forms of this disease are not yet available. Here in this talk, I will discuss our recent study that employs the lipid nanoparticles (LNP) for targeted inhibition of a clinically relevant sphingolipid metabolic pathway in the hepatocytes with siRNA. With this LNP-siRNA approach, we achieved effective lowering of both hepatic and circulating ceramides and improved therapeutic indexes in animal models of non-alcoholic fatty liver disease. Apart from biochemical and histological evidence, the therapeutic efficacy was also confirmed by our recently developed myeloperoxidase-responsive MRI imaging. This proof-of-concept study demonstrates the feasibility of LNP siRNA system for the treatment of metabolic diseases.

15:30

Mid-Afternoon Coffee Break and Networking in the Exhibit Hall


Session Title: Developing and Commercializing Therapeutics from EVs -- QA/QC Metrics and Challenges

16:00

Mario GimonaKeynote Presentation

The EVs and the Secretome: Challenges and Obstacles with EV-based Therapeutics Development
Mario Gimona, Head of Manufacturing, GMP Unit, Paracelsus Medical University Salzburg, Austria

Vesicle-based therapy is increasingly being pursued as a safe, cell-free strategy to combat various immunological, musculoskeletal and neurodegenerative diseases. Extracellular vesicle (EV)-enriched preparations obtained from multipotent mesenchymal stromal cells (MSCs) are of particular interest for therapeutic use since they may convey anti-inflammatory, anti-scarring and cyto-protective activities to the recipient cells and tissues. MSCs secrete a variety of bioactive autocrine and paracrine factors including cytokines, chemokines, extracellular matrix proteases, EVs and growth factors. The MSC-derived secretome is thus a complex mixture of proteins, lipids, macromolecular assemblies and vesicular structures, including various types of EVs. The current knowledge on the dynamic structure of vesicular secretome and its interrelated functions is fragmented and the understanding of the nano- and mesoscale properties is limited. The heterogeneity of the vesicular secretome preparations hampers dose finding, the importance of EV uptake and natural tropism remains to be solved, and technologies to better investigate secretome dynamics and corona/halo formation must be developed. Importantly, activities must be initiated that attempt to quantitatively describe one or more biological activities present in/on EVs that are beneficial in elucidating the complex mode-of-action of the vesicular secretome and to assist in the required dose-finding for pre-clinical and clinical application.

16:30

Eva RohdeKeynote Presentation

Steps to Translate EV-Based Therapeutics into Clinics
Eva Rohde, Head of Department for Transfusion Medicine, Director of GMP Laboratory, Paracelsus Medical University Salzburg, Austria

Extracellular vesicles from mesenchymal stromal cells (MSC-EVs) are being investigated as novel promising biologic drug candidates. In her talk, Eva Rohde will highlight the pharmaceutical and clinical development of naïve umbilical cord-derived MSC-EVs as candidate therapeutics since about 10 years. Therapeutic concepts for selected target diseases are based on observed anti-inflammatory, anti-fibrotic, and neuroprotective biological activities of MSC-EVs in various in vitro and in vivo models. Non-clinical safety and efficacy data required for clinical evaluation include pharmacodynamic, pharmacokinetic and toxicological results for the selected routes of administration and specific indications. Pilot clinical experiments to confirm the EV-associated prevention of secondary tissue damage following acute traumatic injury or chronic (neuro)degeneration and the developmental road for first-in-human clinical trials are presented.

17:00

Xcell Therapeutics Inc.Animal-Origin-Free Chemically Defined Media for Human MSC and Serum-Free Chemically Defined Media for Exosome Isolation: Revolutionary Advances in Stem Cell and Exosome Research
Hyungtaek Jeon, Global Product Manager, Xcell Therapeutics Inc.

Xcell Therapeutics introduces two groundbreaking products that aim to revolutionize human mesenchymal stem cell (hMSC) research and exosome isolation. The first product CellCor™ MSC CD AOF is Animal-Origin-Free Chemically Defined Media for Human MSC, which significantly improves cell proliferation, reduces senescence, and demonstrates minimal variation compared to fetal bovine serum (FBS) media and other Chemically defined media.

The second product is a serum-free, chemically defined medium developed for the expansion and isolation of hMSC-derived exosomes called for CellCor™ EXO CD. This innovative medium offers safety, efficiency, stability, and purity, producing higher volumes of extracellular vesicles (EVs) with reduced background particles. Additionally, our novel approach allows for efficient recovery of high-potency exosomes without the need for media change, thereby optimizing exosome collection. Overall, these breakthrough products have the potential to drive advancements in stem cell and exosome research, facilitating discoveries and applications in regenerative medicine and therapeutics.

17:30

Bernd GiebelKeynote Presentation

Clinical Potential of MSC-EVs and Translational Challenges
Bernd Giebel, Group Leader, Institute for Transfusion Medicine, University Hospital Duisburg-Essen, Germany

18:00

Networking Reception with Dutch Beer -- Network with Colleagues and Engage with Exhibitors

19:00

Close of Day 1 Conference Programming

Tuesday, 14 November 2023

08:00

Morning Coffee and Networking in the Exhibit Hall


Session Title: Challenges and Opportunities in the LNP Space


Session Chair: Ray Schiffelers - UMC Utrecht

09:00

Raymond SchiffelersConference Chair

Synthetic Lipid Nanocarriers: From Liposomes to Lipid Nanoparticles
Raymond Schiffelers, Professor of Nanomedicine, University Medical Center Utrecht, Netherlands

2023 marks the sixtieth birthday of ‘liposomes’. These structures were shown to spontaneously form when amphiphilic lipids were brought in an aqueous environment. In the next decades, many lessons were learned on the biomedical applications of liposomes on the effects of liposome size and surface charge, on steric stabilization and lipid composition, on targeting and immune evasion. Tens of liposome-based products are currently marketed or in advanced stages of clinical development. The most recent addition to the clinically used nanocarriers are the lipid nanoparticles for the delivery of nucleic acids. Building on the foundations of liposomes, these newest type of particles have been successfully used in the siRNA-product patisiran. This recipe also formed the basis for the mRNA COVID-vaccines. These successes paved the way for many new applications where synthetic lipid nanoparticles enable new nucleic acid modalities for advanced therapeutic interventions.

09:30

Fred CampbellKeynote Presentation

LNP 2.0+: Gene Medicine Beyond Covid
Fred Campbell, Scientific Director, NanoVation Therapeutics UK, United Kingdom

mRNA vaccines against SARS-Cov2 have emphatically shown lipid nanoparticles (LNPs) can deliver safe and effective gene therapies in humans, can be developed at unprecedented speed, manufactured at scale and all at reasonable cost. A genetic healthcare revolution is now upon us. How quickly these new, diverse and life changing treatments come to market, however, will require many outstanding challenges to be met (both scientifically and commercially). Here, I will briefly outline NTx’s strategic and scientific efforts to accelerate the development of new LNP-based gene medicines and democratize healthcare for all.

10:00

Nan ZhangKeynote Presentation

Screening and Scale-Up Lipid Nanoparticles Using a High-Throughput Microfluidic System
Nan Zhang, Associate Professor, University College Dublin, Ireland

Drug delivery systems in forms of nanoparticles have a broad spectrum of applications in area of gene therapy, cell therapy, and vaccine development. Lipid nanoparticles (LNPs) or liposomes that consist of a lipid bilayer and have a hollow structure are the most widely used drug carriers for nanomedicines. It is possible to encapsulate a variety of low molecular compounds (e.g., drugs, peptides, antibody, plasmid-DNA, mRNA, siRNA etc.) in the lipid bilayer membrane or to encapsulate them into the hollow structure. However, significant concerns remain on efficacy, safety, consistency, scale-up of manufacturing and stability when translating LNPs from formulation into clinical application. LNPs’ properties, e.g., size, size distribution, charge, drug encapsulation, transfection efficiency etc., can significantly affect the bio-distribution and pharmacokinetics of the drug to be delivered. In order to obtain LNPs with expected characteristics, microfluidics that exploit fluidic control to synthesize LNPs have grown in acceptability and applications in laboratory and larger scales due to offering a reproducible and robust manufacturing through the precise control of flowing conditions. In the present talk, the author will share the newly developed high throughput microfluidic system for laboratory screening and formulation of nanoparticles based on a novel aerofoil microfluidic mixing system. The formulation based on low, medium and high flow rates will be demonstrated. The automation of the system will be introduced for industrial scale formulation. The device has demonstrate the strong potential for low volume screening, medium volume scale up and high volume GMP production for nanoparticle development.

10:30

Mid-Morning Coffee Break and Networking in the Exhibit Hall

11:00

Scale-up Development and Manufacturing of Nanomedicines: From Tech Transfer to Clinical Batch
Chiara Pretto, Senior Scientist, Process Development, Ardena Nanomedicines, Netherlands

Nanomedicines represent the cutting-edge frontier in the pharmacological field, providing unique solutions for the treatment of various diseases as well as for imaging purposes. The rapid advancement in this field poses significant challenges to clinical development and requires a carefully planned development scheme executed by scientists experienced in nanoparticle synthesis and characterization. As a contract development and manufacturing organization at the forefront of the nanomedicine landscape, Ardena embraces these challenges and can provide fine-tuning of manufacturing techniques, development of tailor-made reproducible, scalable, and GMP-amenable processes as well as full characterization of products. We will present how we navigate our customer’s lead nanomedicines through drug development, and we will highlight some of the challenges in bringing nanoparticles-based therapeutics from discovery to the clinic.

11:30

The EVENEW Study - From the Production Process Development to the Clinic
Marcin Jurga, Chief Scientific Officer, EXO Biologics SA, Belgium

EXO Biologics received approval from the EMA for the EVENEW study, an adaptive, seamless Phase 1/2 trial assessing the safety and efficacy of intratracheal administration of EXOB-001 in preventing Bronchopulmonary Dysplasia (BPD) in preterm newborns. The EVENEW study is the first mesenchymal stromal cell-derived extracellular vesicles (EVs) trial approved by the EMA.

The production platform and the manufacturing process developed by EXO Biologics were crucial in clinical translation of EXOB-001. The platform was designed to achieve a reproducible and efficient production of the EVs. The platform is broadly scalable maintaining the drug product’s identity.

The production process is essential in the Company’s development strategy focused on novel drug candidates for therapeutic applications in Respiratory Diseases, Gastroenterology, Neurology, and drug delivery.

12:00

Polyplus-TransfectionCationic Lipids Offer New Possibilities in Liposome and LNP Engineering for RNA Therapeutics
Jean-Thomas Issenhuth, Key Account Manager EMEA, Polyplus-Transfection

Lipid-based nanoparticles (LNPs) represent one of the most promising RNA delivery systems, not only due to their efficacy, but also to the ease of production achieved through precise controlled mixing technique known as microfluidics and the scalability of production. Still, they present some limitations related to toxicity, targeting, limited biodistribution through systemic administration which mainly results in liver uptake. Polyplus® has recently developed a library of cationic lipids (LipidBrick®), which represents a remarkable innovation compared with conventional cationic lipids by ensuring reduced toxicity of LNPs and extending their use beyond the liver.

12:30

Networking Buffet Lunch in the Exhibit Hall -- Networking with Colleagues, Engage with Exhibitors

13:30

Malvern PanalyticalReveal the Nanoworld with NanoSight Pro
Agnieszka Siupa, Product Manager, Malvern Panalytical

Developing therapeutics using invisible drug delivery systems brings many challenges in biophysical characterization. During process and formulation development, there is a need to monitor sample purity and stability at many points to help guide the next step. Garnering detailed information of these complex systems to fully correlate their composition to their therapeutic effect is also crucial. NanoSight Nanoparticle Tracking Analysis (NTA) has become a standard toolbox in the characterization of biomaterials including extracellular vesicles, drug delivery systems, viral vectors and protein aggregates. NTA provides visual confirmation and high-resolution size and concentration data in minutes allowing the instant assessment of sample stability, but also complexity. With the introduction of the new NanoSight Pro, characterization becomes even easier, quicker and more accurate than before. Packed with smart features, the NanoSight Pro provides greater sensitivity in biologics detection, high reproducibility and enhanced fluorescence measurements for detecting sample subpopulations. Powered by machine learning algorithms, measurement subjectivity is reduced and automated processing enabled to assure superior Nanoparticle Tracking Analysis.

14:00

Roy van der MeelKeynote Presentation

Gene Silencing in HSPCs with an Apolipoprotein A1-based Nanodelivery Platform
Roy van der Meel, Assistant Professor, Department of Biomedical Engineering, , Eindhoven University of Technology, Netherlands

Nucleic acid therapeutics are revolutionizing healthcare via gene inhibition, addition, replacement or editing. However, nucleic acid-based drugs require chemical modifications and sophisticated nanotechnology to prevent their degradation, reduce immunostimulatory effects, and ensure intracellular delivery. Lipid nanoparticle (LNP) technology is the current gold standard platform that enabled the clinical translation of the first siRNA drug Onpattro and the COVID-19 mRNA vaccines. Nevertheless, current LNP systems are mostly suited for vaccine purposes following local administration or hepatic delivery following intravenous administration. To unleash RNA’s full therapeutic potential, we introduce modular nanoplatform technology for systemic nucleic acid delivery to immune cells in hematopoietic organs using apolipoproteins.

14:30

Rut BesselingKeynote Presentation

Process Analytics for Nanotherapeutics Manufacturing, Enabled by Novel Inline Nanoparticle Sizing Technology
Rut Besseling, Scientific Director, InProcess Instruments BV, Netherlands

The rise of nanotherapeutics -and nano based products in general- has increased  the need for better characterization, understanding and control of nano-particle (NP) suspensions in synthesis and processing. In Pharma this is emphasized by regulatory drives for Quality by Design (QbD) and process monitoring via Process Analytical Technology (PAT). But continuous measurement of NPs like LNPs or liposomes using PAT integrated in the production process, has so far hardly been possible. Specifically, NP size characteristics -critical for endproduct quality- could previously not be measured inline due to the challenges regarding e.g. suspension turbidity and ‘agitation’ for standard methods (e.g. standard Dynamic Light Scattering, DLS). This has been an important barrier in successful development, scale-up and manufacturing of nanotherapeutics. Here I will describe applications of a new non-invasive PAT instrument – the NanoFlowSizer (NFS)- providing continuous, real-time, in-process size characterization of NP suspensions, over a vast range of suspension turbidities and flow conditions. The NFS resolves scattered light signals as function of depth in the suspension (‘Spatially Resolved Dynamic Light Scattering’), which provides the key solution to the turbidity/flow challenges mentioned for inline NP sizing. After a brief technology intro, several examples are described highlighting the NFS’ novel process monitoring capabilities. These include monitoring of solid NP syntheses (SiO2/TiO2/Ibuprofen), pharmaceutical nano-emulsion homogenization, processing of protein solutions and, lastly, continuous manufacturing of liposome formulations, including unique PAT-based automated process control for ‘on target’ production.

15:00

Sai Kiang LimKeynote Presentation

Anti-Aging MSC-EVs
Sai Kiang Lim, Research Director, Institute of Medical Biology, A*STAR, Singapore

Aging is believed to be, in part, a result of the decline in functional adult stem cells that are crucial for maintaining tissue homeostasis. We had previously postulated that MSC EVs could help preserve the balance of cell numbers by activating survival signaling pathways. Indeed, our research showed that EVs derived from MSCs effectively reduced senescence both in cell cultures and in live organisms, leading to an improvement in the natural lifespan of aging mice.

15:30

Mid-Afternoon Coffee Break

16:00

Round-Table Open Discussion Focusing on EVs and LNPs for Therapeutics Development -- Chaired by Dominique de Kleijn and Raymond Schiffelers

17:00

Close of Conference