Complex cellular-HTS based assays for modulation of tumor microenvironment and metastasis

CIPF is a specialist screening site focused on complex cellular assays. Within this call, CIPF offers a High Throughput Screen on complex cancer cell models (including co-cultures, spheroids and organoids, from commercially available cell lines up to patient derived samples). CIPF offersexpertise from assay miniaturization up to hit confirmation, based on assays in 96 or 384 well plates with quantification of absorbance, fluorescence, luminescence, or High-Content Fluorescent Imaging among others.

Outputs of the HTS assays include monitoring cell viability (e.g., MTS, LDH, ATP) or gene expression, proliferation-invasiveness markers, cell death (caspase assays), cell cycle (CDKs), inflammation (alpha screen technology or luminex), intracellular rearrangements (BRET, FRET) as well as tumor-derived exosome modulation: protein expression quantification by Immunofluorescence (In cell analyzer) and alpha screen (exoscreen).

CIPF will offer screenings to find bioactive molecules in complex cellular assays (intracellular rearrangements), co-cultures, organoids and primary cultures. HTS Assays for modulation of tumor microenvironment and metastasis including Synergism studies. The CIPF screening platform was established in 2005. The platform is composed of a highly multidisciplinary team which incorporates expertise in chemistry, cellular and molecular biology, medicine, pharmacy and bioinformatics and has a broad experience in the miniaturization of complex cellular assays and their adaptation to HTS format. The CIPF screening site is fully equipped to work with HTS screening systems and offers a broad expertise in working with different type of cells from commercially available to primary culture cells (murine and human derived- 2D and 3D spheroids and organoids including patient derived samples for cancer applications) not only in cancer but also in cardio and neuro-derived complex models and even orthotopic cultures (human skin).

Within this call, CIPF offers a High Throughput Screening on those more complex cancer cell models described above, from miniaturization up to hit confirmation, based on assays in 96 and 384 well plates with quantification of absorbance, fluorescence, luminescence, or High-Content Fluorescent Imaging among others. Outputs of the HTS assays include monitoring cell viability (e.g., MTS, LDH, ATP) or gene expression, proliferation-invasiveness markers, cell death (caspase assays), cell cycle (CDKs), inflammation (alpha screen technology or luminex), intracellular rearrangements (BRET, FRET) as well as tumor-derived exosome modulation: protein expression quantification by immunofluorescence (in cell analyzer) and alpha screen (exoscreen). Drug synergism studies (combination therapies: drug ratio, drug PK) can be offered.

Number of compounds screened

100,000 (full commercial set) of the EU-OPENSCREEN ERIC compound library or 5,000 (for highly complex assays or those based on patient derived samples (3D organoids)).

Time frame for screen up to validated hits

6-8 months: depends on the development stage of the assay and the compatibility of the bioassay with the screening system.

CIPF laboratories are fully equipped to work with HTS screening systems (FreedomEvo Workstations LiHa+MCA96 ARM (Tecan) for 96-384 plates and dispose of highly advanced screening technologies: Multitask plate readers: EnSigth (Perkin Elmer), Clariostar (BMG LabTech), CytoFlex (Beckman Coulter), cytomics FC500 MCL, InCell Analyzer 2000, Magpix (Luminex technology), In vivo imaging (LeicaDM6000I, LeicaTCS-SP2-AOBS, LeicaTCS-SP8-Hyvolution II+ HCS multiplate). The drug discovery platform at the CIPF offers a broad spectra of readout technologies (absorbance, anisotropy, fluorescence, BRET, FRET;TRF, TRF-FRET, label-free, alpha screen, confocal image-based, flow cytometry image-based etc.).

• Validation of the assay provided by the user to ensure robustness (Screening system).
• Miniaturization of the assay into 384-well plates (if applicable) (Compound management and Screening system).
• Pilot screen using 5,000 compounds from the EU-OS compound library (Compound management system and Screening system).
• Analysis of pilot screen data (data management system).
• Full screen with the remaining 95,000 compounds of the EU-OS compound library (compound management system, screening system, microbiological and cell biological infrastructure).
• Data analysis and subsequent selection of primary hits.
• Hit picking of primary hits for hit validation.
• Hit validation by variation of the compound concentration (determinations of IC50- or comparable parameters); eventually performance of relevant counter screens (e.g., effects on reporter proteins).

Please note that this project includes the possibility of re-screening during potential chemical optimization under the medicinal chemistry call starting in 2021 (tentative date).

The prerequisite starting point to qualify for access is the availability of an established bioassay and associated key bespoke reagents, developed at lab-scale by the successful applicant. The assay format is compatible with the performance in microtiter plates allowing the quantitative determination of an optical parameter (e.g., absorbance, fluorescence, luminescence). 

Positive and negative controls are identified, and the resultant quantitative assay data are sufficiently different to allow unambiguous distinction.The target of the assay is determined by the applicant’s bioassay.

As specifics of the assay transfer procedure may vary between partner sites, the applicant and the individual sites will agree on the appropriate steps and logistics together.

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Topics / Keywords:

drug discovery, small molecules, high-throughput screening, cell-based assays

Open Time:

Start date:
June-01-2019 (20:00 CET)

Closing date:
September-30-2019 (20:00 CET)

Contacts:

Scientific Contact-Coordinator: Dr. María J.Vicent, mjvicent@cipf.es

Scientific Contact: Dr. Mar Orzáez, morzaez@cipf.es

Technical Contact: Esther Masià, emasia@cipf.es