Ariadne platform : High throughput and high content screening

Plateformes Lilloises en Biologie & Santé – PLBS – UMS 2014 – US 41

Plateforme Ariadne Pasteur Lille

Présentation

The ARIADNE-screening platform includes two screening infrastructures for small molecules and SiRNA, both located on the campus of Institut Pasteur in Lille:

  • The first system, dedicated to HTS (high throughput screening), enables screening automation and multimode reading.
  • The second robotic system of HCS (high content and high throughput screening) allows to perform multiparametric cell screens, based on the detection of photons.

Know-how and services offered :

The platform brings together a set of expertise, know-how and equipment dedicated to the screening of biological disruptors such as small chemical molecules or siRNA libraries. The equipment chain allows full or partial automation of the screening steps, from the plating of disruptors from chemical libraries or siRNA libraries, to the reading of results. The know-how and technical skills of the personnel allow the development of high-throughput miniaturized (96, 384-well plates) and automated tests for in vitro, in bacterio, or in cellulo, target or phenotypic screening. ARIADNE benefits from access to chemical libraries, in particular the regional chemical library of approximately 200,000 chemical molecules, siRNA and miRNA libraries. The platform also has expertise in cell labeling for cell domiciliation, using CRISPR Cas9 approaches. 

Equipment are located in biosafety level 1 to 3 laboratories. ARIADNE-HTS in BSL-1 allows management of libraries and high-throughput screening on multimode plate readers (with measurements in fluorescence, luminescence, absorbance, etc…). ARIADNE-HCS, from the ImagInEx BioMed Equipex, use automated confocal microscopy methods. A first imaging system (IN cell Analyzer 6000 GE Healthcare) is fully automated in BSL-2. A second system (Opera® HCS System-PerkinElmer) is available in BSL-3. This so-called “non-destructive” technology allows the monitoring of the same sample over time both in the short term (dynamic phenomena) and in the long term (kinetics) and makes it possible to measure several parameters simultaneously on the same biological sample. 

Numerous tests can be implemented: Enzymatic tests, phenotypic or target cell tests, protein-protein interactions in vitro or in cellulo, Thermal Shift Assay (TSA) and monitoring of target engagement in cellulo by CETSA… The use of “Echo” acoustic technology for liquid nanotransfer makes it possible to test molecules in combination and in dose response. 

As part of a partnership agreement, projects can be continued with the experts of Ariadne-Criblage towards the characterization of targets and the optimization of modulators: target identification and engagement, medicinal chemistry, measurement of physicochemical properties, ADME studies and pharmacokinetics in rodents.

Highlights

ARIADNE technicians, engineers and researchers are committed to finding a treatment for the Covid-19 pandemic

  • A molecule active in vitro on SARS-Cov-2 infection was discovered by cell screening of a library of 2000 drugs on the HCS system in BSL-3. This discovery is the result of collaboration with Drs Sandrine Belouzard  and jeran Dubuisson from the Lille Center for Infection and Immunity and researchers from the company Apteeus.
  • The HTS screening of the entire ARIADNE library (approx. 100 000 compounds) has identified several chemical series active on the 3CLPro protease of SARS-Cov-2 at sub-micromolar concentrations. This project is being carried out with Dr Julie Charton from Unit U1177.

Members

The Ariadne-screening platform is part of the UMS 2014-US41-Platforms Lilloises in Biology and Health and of the National Research Infrastructure ChemBioFrance.

Its steering committee brings together researchers from the Lille Infection and Immunity Center : Dr Priscille brodin and Dr Alain Baulard, as well as from Unit U1177 : Pr Benoît Déprez and Dr Florence Leroux.

Priscille BRODIN
DR Inserm, CIIL UMR9017 U1019

Benoit DEPREZ
Pr Univ Lille, U1177

Alain BAULARD
DR Inserm, CIIL UMR9017 U1019

Florence LEROUX
IR Inserm, U1177

Alexandre VANDEPUTTE
IE Univ Lille, CIIL UMR9017 U1019

Cyril COUTURIER
MCU Univ Lille, U1177

Valentin GUILLAUME
CE IPL, U1177

Julie DUMONT
AI Univ Lille, U1177

Nathalie DEBOOSERE
CE IPL, CIIL UMR 9017 U1019

Valérie LANDRY
CE IPL, U1177

Adrien HERLEDAN
IE Inserm, U1177

Sandrine WARENGHEM
Tech IPL, U1177

Publications

Lesire, L., Chaput, L., Cruz De Casas, P., Rousseau, F., Piveteau, C., Dumont, J., Pointu, D., Déprez, B., and Leroux, F. (2020).
High-Throughput Image-Based Aggresome Quantification.
SLAS Discov 25, 783-791.

Moraski, G.C., Deboosère, N., Marshall, K.L., Weaver, H.A., Vandeputte, A., Hastings, C., Woolhiser, L., Lenaerts, A.J., Brodin, P., and Miller, M.J. (2020).
Intracellular and in vivo evaluation of imidazo[2,1-b] thiazole-5-carboxamide anti-tuberculosis compounds.
PLoS One 15, e0227224.

Herledan, A., Andres, M., Lejeune-Dodge, A., Leroux, F., Biela, A., Piveteau, C., Warenghem, S., Couturier, C., Deprez, B., and Deprez-Poulain, R. (2020).
Drug Target Engagement Using Coupled Cellular Thermal Shift Assay-Acoustic Reverse-Phase Protein Array.
SLAS Discov 25, 207–214.

Blondiaux, N., Moune, M., Desroses, M., Frita, R., Flipo, M., Mathys, V., Soetaert, K., Kiass, M., Delorme, V., Djaout, K., Trebosc, V., Kemmer, C., Wintjens, R., Wohlkönig, A., Antoine, R., Huot, L., Hot, D., Coscolla, M., Feldmann, J., Gagneux, S., Locht, C., Brodin, P., Gitzinger, M., Déprez, B., Willand, N., and Baulard, A. R. (2017).
Reversion of antibiotic resistance in Mycobacterium tuberculosis by spiroisoxazoline SMARt-420.
Science 355, 1206–1211.

 

Team contact

Florence Leroux
Scientific group leader

florence.leroux@pasteur-lille.fr