We were present at the last HCS conference in Boston and, during these two days of intensive discussions, lots of topics were addressed in HCS and HCA. I summarize below what seems really important for me.

A lot of molecules fail in the process of drug development in a very late stage (more than 50% in pre-clinical step and phase I for safety reasons, and 80 % for efficacy in phase II). Furthermore, there is more and more data showing that there is a huge transitional problem between animal and human studies. During this event, the pharmaceutical industry and academic groups have shown the importance to use phenotypic screening to found new drugs, but with improved relevance of the in vitro models and data analysis as explain, for exemple, by Christophe Antczak from Novartis. They have published a review on this topic : “How Phenotypic Screening Influenced Drug Discovery: Lessons from Five Years of Practice“. Kristin Fabre from AstraZeneca and Matt Wagoner from Takeda Pharmaceuticals have also highlighted the need for better human models for drug research, in order to improve efficacy and safety of molecules during the early steps of drug research. Another review was presented by Kristin Fabre from AstraZeneca, explaining why we need better human models : “Lessons learned from the fate of AstraZeneca’s drug pipeline: a five-dimensional framework.

To improve relevance of the in vitro models, many presentations during this event were about 3D culture and the involvement of the micro-environment with modification of the extracellular matrix (ECM). As explained by Sophie Lelièvre from Purdue University for cancer research, during the tumoral progression, the ECM is modified with an increase of its density due to collagen production from cancer cells. The density of the extracellular matrix is a hallmark of tumor aggressiveness. During this tumor progression, there is an increase in the heterogeneity of the cells in the different grades, inducing resistance to chemotherapy. To mimic the tumor progression in in vitro models, it is necessary to reproduce this cellular heterogeneity with the modification of the extracellular matrix in 3D culture.

To improve the relevance of cellular models, microfluidics are also more and more used to generate dynamic models and to better mimic in vivo situation. Different presentations about “organ-on-a-chip” systems during this event have shown that this technology is getting more and more advanced. Jonathan Himmelfarb, from the University of Washington, has shown that 3D model including microfluidic devices to mimic human kidney proximal tubule, express higher transporters and metabolic enzymes compared to 2D model on Transwell filters. Finally, this technology has been already transferred from academic labs to the pharmaceutical industry as shown by Kriten Fabre from AstraZeneca. For example, they use microfluidic systems to check the potential inflammation of lipid nanoparticles.

This event was really of great interest to show that all new technologies like 3D culture, bioprinting, microfluidics for cell culture – and deep learning/machine learning and AI for data analysis – are increasingly used and synergistically help to get predictive human models.

Review from Novartis : http://online.liebertpub.com/doi/10.1089/adt.2017.796

Review from AstraZeneca : http://dx.doi.org/10.1038/nrd4309

I will be present at the High content Analysis and 3D Screening Conference in Boston from Monday (November 7)  to Wednesday (November 9).

“Now in its fifteenth year, Cambridge Healthtech Institute’s High-Content Analysis & 3D Screening Conference will deliver the most recent developments in high-content and phenotypic screening. Leading pharma and academic researchers will focus on advancements in HCA technologies and applications, including screening of 3D and physiologically-relevant cellular models, data analysis techniques, and case studies and strategies for successful drug discovery. The expanded 3-day coverage on 3D cellular models and 3D screening will present the latest in spheroid, organoid and organotypic cell culture, and organ-on-a-chip technologies for drug screening, toxicity testing, and disease modeling.”

In this conference, I will present a poster on our collaborative work with Dr Karim Si-Tayeb, researcher from Institut du Thorax (CHOPIN project) on differentiation of iPS in hepatocytes in 3D culture for metabolic diseases screening.

If you want to know more on this project, don’t hesitate to come and discuss with me or contact us.

As last year, we will be very happy to attend Drug Discovery 2017 event in Liverpool (Innovation Zone, IZ1 booth). We will present our activities, the new assays that we have developed recently and our brand new robotic platform.

“Drug Discovery is ELRIG’s flagship event. Now in its 11th year, we return to ACC in Liverpool to bring together the leading lights in drug discovery research across pharma, biotech, academia and the vendor community.”

As you can see on the dedicated webpage, the event program is highly interesting:

  • Advances in Imaging
  • Drug Discovery in the 4th Dimension
  • Innovations in Assay Design, Development and Screening

And of course, don’t forget the amazing “Innovation Zone”, the right place to discover future partners, as HCS Pharma !

Come and join us in Drug Discovery 2017 in the Innovation Zone at booth IZ1, to discuss with Nathalie, our CEO and Pierre-Jean, our in vitro toxicology expert !

Nathalie will be present tuesday 3 October, 3-4 pm, on booth of our partner Molecular Devices (#D18) to answer your high-content screening (HCS) questions.  Book a time slot to meet Nathalie !


Great start for Eurotox 2017 ! Congress has begun with a lovely opening ceremony with traditional slovakian dancers. The first keynote of professor Philip Steven was a passionated overview of neurosciences between non-human and human research, performed in neurovigil team between NASA and MIT.

I am very excited to attend the following sessions! If you want to meet me, don’t hesitate to contact me!

During these last two weeks, we took control of our new robotic platform and validated the pipetting volumes and the entire process by testing it with a few plates. Some steps are still to be improved, such as the scheduling of the plates. The first screening with a few plates was on hepatotoxicity assay on SCREEN-WELL® Hepatotoxicity library from ENZO (238 compounds) in one concentration.

After improving our process, we will test this library in dose effect in addition to the Prestwick library. We will also perform neurotoxicity assay in a screening mode on internal and Prestwick library. These assays will allow us to validate all our process to cell culture until data analysis in addition to exemplify data on our toxicity assays.

Cell biology is part of the R&D activity in dermocosmetology, as providers of active extracts or pure compounds have to demonstrate their efficacy and safety. At HCS Pharma, we are developing new dermocosmetology assays on relevant cellular models, like human primary keratinocytes and human primary fibroblasts, using our automated platform and high content analysis system. Through the use of 96 or 384 wells plates combined with process automation, we constantly increase the throughput in order to allow proof of concept assays of one or several compounds up to the screening of large compounds libraries, for single endpoint or live cells analysis to multiplexed phenotypic screening.
Assessment of compounds activity/safety can be performed on several parameters in parallel with a multiplexed assay. Among them, we describe here wound healing, inflammation assay & extra cellular matrix analysis, which are routinely performed. Other parameters can be analysed on demand on live and fixed cells, using chemical probes or immunocytochemistry.

View and download on Slideshare (low quality) : https://www.slideshare.net/hcspharma/in-vitro-dermocosmetology-high-content-analysis-approach-using-human-primary-keratinocytes-and-fibroblasts

Ask for high quality link  by putting your email below

I enjoyed to participate to this event hold in Stevenage at GSK, in a beautiful and impressive setting. The combination of uses of new technologies as high content imaging (HCA/HCS), imaging mass-spectrometry or imaging flow cytometry and new data analysis as machine learning and deep learning were presented in this event. These combinations show an extremely nice perspective in pharmaceutical research and in personalized medecine. I was really impressed by the quality of the presentations. James Pilling (Aztrazeneca) showed cardiotoxicity assay on 3D spheroid by using HCS technology. Emmanuel Gustin (Janssen) presented phenotypic screening by using phingerprints and machine learning. Rod Benson (Imagen Therapeutics) talked about precise and personnalised medecine for cancer by using HCS technologies.

All these emerging technologies and data analysis systems promise interesting discoveries in the near future.

Méryl Roudaut* will present a poster on the work of Institut du Thorax (Nantes, France) during the meeting “advances in cell engineering, imaging and screening conference”. This conference will be held on 17-18 november 2016, in Louvain (Belgium). It will focus on novel technologies through presentations in several sessions:

  • New imaging and microscopy tools
  • Super-resolution Imaging
  • IPS and cell reprogramming
  • 3D cell culture and organoids
  • Cell manipulation
  • In vivo cell-based assays
  • Cytometry
  • High Content Screening.

If you want more informations about the CHOPIN program and the work presented on the poster, don’t hesitate to discuss with Méryl* during this event!

*Méryl is our PhD student working in collaboration with Pr Bertrand Cariou and Dr Karim Si-Tayeb within the CHOPIN RHU program.

We are always happy to read reports showing the growing interest of the research for high content screening. For those interested, you can find in this article to appear, an expert opinion on High Content Screening in the drug-discovery processes. The team of Fraietta & al(1) rated the number of semantic researches on the HCS on Pubmed. They detail the therapeutic areas covered by the HCS (neurological diseases, cardiovascular diseases, infectious diseases, oncology …), its role in the drug discovery process, but also all its future potential developments. Among these, one finds, in addition of the use of primary cells culture:

– The use of iPSCs[i],

– The adjustment of the technique for 3D cell cultures, tissues, organoids and even whole organisms (unicellular organisms like Saccharomyces cerevisiae, or multicellular organisms like the zebrafish),

– The development of engineered cell lines with genome editing techniques (to introduce a mutated driver oncogene, or to silence a tumor suppressor gene) including homologous recombination, approaches based on ZNF[ii], TALENs[iii] or CRISPRs[iv]/Cas9.

In the end, such an article confirms our choice of HCS as Core Development technology.

Opportunities for HCS

 

  1. Fraietta I, Gasparri F. The development of high-content screening (HCS) technology and its importance to drug discovery. Expert Opin Drug Discov. mai 2016;11(5):501-514.

 

[i] iPSCs = induced pluripotent stem cells

[ii]  ZNF = zinc-finger nucleases

[iii] TALENs = transcription activator like effector nucleases

[iv] CRISPRs = Clustered regularly interspaced short palindromic repeats

 

HCSPharma-Galderma

During ELRIGfr event in Rennes, We had the opportunity to present our work on genotoxicity assay (comet assay on TK6 & gH2AX on keratinocytes and HepG2) in collaboration with the team of in vitro of Galderma R&D. It is really exciting to work with them since they are really experts in their domain and they want to work on innovative solutions in HCS. We really want to thank them for our interesting discussion and our collaborative projects!

HCSPharma-Galderma

PosterGenotoxHCSPharma-Galderma

Tincosmetics-2016he annual high mass of the European In-Cosmetics event is now over for 2 weeks. We are sincerely glad to have met so many interesting people. We were also very excited to show in further details the place of HCS (High Content Screening) in active ingredients discovery process, and to speak of its effectiveness and its relevance during the R&D process. Feel free to let us know your draft studies and we will work hand in hand to provide you a suitable protocol, with quantified results as well as striking pictures.

We also take advantage of this post to thank BCI (Bretagne Commerce International) and CBB (CapBiotek) for their constant support as well as during the show.

Latest news: Come and join us next 3rd and 4th May in Rennes for the free ELRIGfr congress on Lab Automation. We will present 3 posters on genotoxicity and neurotoxicity.

We look forward to seeing you there!

Cell-based high-content screening (HCS) assays have become an increasingly attractive alternative to traditional in vitro and in vivo testing in pharmaceutical drug development and toxicological safety assessment. The time- and cost-effectiveness of HCS assays, combined with the organotypic nature of human induced pluripotent stem cell (iPSC)-derived cells, open new opportunities to employ physiologically relevant in vitro model systems to improve screening for potential chemical hazards. In this study, they used two human iPSC types, cardiomyocytes and hepatocytes, to demonstrate the potential of a variety of HCS assay combinations for testing the potential toxicity of chemicals and complex substances. The overall aim was to improve in vitro toxicity testing by reducing the time and cost of the assays while enhancing the mechanistic interpretation of the readouts so that confidence in animal replacement tests is improved.

To know more, read this article: http://www.genengnews.com/insight-and-intelligence/high-content-assay-multiplexing-for-toxicity-screening-in-induced-pluripotent-stem-cell-derived-cardiomyocytes-and-hepatocytes/77900591/

“Manufacturers have developed super-corrected optics that allow co-localization studies with much more accuracy than before”, said Kevin Jia, senior marketing manager of Life Science at Olympus Scientific Solutions Americas in Waltham, Mass. “Also, throughput on our systems is much higher. Spectral imaging is vastly improved and live-cell imaging is possible with very low laser power.”

Working distance, wavelength ranges and other aspects of the optics now are tailored for specific applications to help confocal move from a fixed-cell technique to a highly sensitive, live-cell technique.

“We see confocal being combined with other imaging techniques,” Jia said. “For instance, we are looking at much higher-resolution and higher-contrast imaging through new superresolution technologies that are incorporated with spinning-disk or other confocal systems to create solutions providing fast, high resolution, high-contrast data.” One example is the Opera Phenix High Content Screening System from Perkin-Elmer, which is a microlens-enhanced Nipkow spinning disk confocal system that allows the capturing of confocal images from samples in multi-well plates or on slides at a very high throughput.

“Compared with conventional confocal laser scanning microscopes (CLSM), the Opera Phenix system allows unsupervised image acquisition, which can produce up to 400,000 images per day,” said Karin Boettcher, associate product manager for High Content Screening and Applications at PerkinElmer, Hamburg, Germany. “This far higher throughput, compared to conventional CLSM, allows screening of whole small molecule or siRNA libraries for a desired phenotypic effect.””

Sentences come from article on photonics.com internet site from Marie FREEBODY, Contributing editor.

To read the full article, go to this link: http://www.photonics.com/m/Article.aspx?AID=58209

Do you know what is phenotypic screening ? If you are here I think yes 🙂 But if you want to explain it easily, I strongly recommend the reading of Perkin Elmer white paper « Phenotypic Drug Discovery with High Content Screening ». It’s short (only 4 pages) but it contents main information to understand differences between « target based » and « phenotypic screening » approaches in drug discovery.

It explains also the importance of the technology side and the digital side of HCS and finally, give a good summarize of our vision :

“Today’s drug discovery strategies require candidate compounds to fail early and cheaply in the discovery stage, rather than late and expensively in the clinical phase. Testing compounds from the beginning in physiologically relevant model systems and leveraging the rich information available in image-based screens are ways to focus on those compounds that give rise to the right phenotypic changes without undesirable effects on the system. High Content Screening with highly detailed multiparametric assays in conjunction with modern machine-learning tools provide a promising way to achieve this goal.”

 Source : http://go.perkinelmer.com/Q315-WhitePaperEmailPKI-INF-LP?cid=4296

I was present at the last congress ELRIGfr in Brussels. It was a strong exchange place with interesting presentations around robotics. It is always really interesting to see examples of innovative automation which give really robust results. For example, Frank Gudermann from university of Bielefield in Germany showed that automation of culture cell with cell count by holographic picture gives more robust results as manual count by trypan blue. Or automation of PK sampling with nanoliter by using echo technology allows increasing throughput of bioanalytical analysis for PK studies in Hoffmann La Roche. Also, another presentation of IGR has shown a complete robotic platform with 4 imagers (Micro from MDS) and 1 FACS totally automated from cell seeding to staining and reading.

It was a great pleasure also for us to present 4 different posters on our few last month works in cell imaging. This work was done in collaboration with Perkin Elmer on the Operetta machine. This machine allows us to take images in a 3D culture as shown in these posters: neuroprotection model for parkinson disease and oncology and performed also really nice images showed in these other posters: hepatotoxicity assay and genotoxicity assays.

Phenotypic screening by High Content Screening monitors all modification of phenotypic changes. The interest of this technology is having a significant content of biological information (phenotypic and target modifications) and this, cell by cell. This important content of information results in a very high amount of data that must be analyzed globally (with PCA reduction for example). Thus, the data analysis and statistics, especially the Z-score that everybody knows, need to be reviewed. Guyon et al boast Φ-score. What do you think about that?

“Using robust statistics and a variance model, we demonstrated that the Φ-score showed better sensitivity, selectivity and reproducibility compared to classical approaches. The improved performance of the Φ-score paves the way for cell-based screening of primary cells, which are often difficult to obtain from patients in sufficient numbers.”

Source : Guyon, L. et al. Ф-score: A cell-to-cell phenotypic scoring method for sensitive and selective hit discovery in cell-based assays.Sci. Rep. 5, 14221; doi: 10.1038/srep14221 (2015)

In this article, authors emphasize the importance of standardisation in the development of HCS. Standards (for Image File Format, Data Integrity and Reference Material) are critical to assure both data quality and reproducibility of assays.

The scientific community must embrace the urgent need for standardization from cells to HCS data to enhance reproducibility and reduce unnecessary economic loss. Together, the scientists and end users partnering with the manufacturing companies, colleagues, and government agencies can strive to build the necessary toolboxes and protocols to ensure the highest quality of all aspects of HCS data.

Source: Standardization of High Content Imaging and Informatics

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