Translation Control Discovery Platform

Translation Control Discovery Platform

Our platform combines breakthrough novel biology with proprietary analysis software that runs in a big-data cloud architecture. Compound libraries are screened in a purpose-built, high performance Translation Control Lab. Using our protein synthesis monitoring assays, we generate millions of images that show the impact of the different compounds on the translation of the target protein. Images are uploaded in real time into the project’s private cloud where they are analyzed by our proprietary bio-informatics, imaging and big data analysis algorithms to identify hit molecules that selectively control the translation of the target protein.

Hit validation and de-risking

Analysis is done in several iterations by applying multiple secondary assays built around our technology. We analyze the impact of hit compounds on overall protein synthesis, eliminating compounds that are toxic, that interfere with the synthesis of many of the cell's proteins or that shut down the translation machinery itself. Additional translation selectivity assays were developed which monitor the effect of compounds on groups of proteins of interest or specific proteins along a pathway.

Hit optimization, Target deconvolution and mechanism of action

The platform verifies the selectivity of hit compounds and offers unique analysis capabilities that assist in optimization, target de-convolution and mechanism of action identification.

Breakthrough Biology

  • Labeled tRNA library

    Fluorescent labeled tRNA library

    The full repertoire of human tRNAs (48 isoacceptors) has been affinity purified and labeled with a fluorescent tag using Anima’s patented technology. Two complete tRNA libraries have been designed: one tagged with an energy donor and one tagged with an energy acceptor. To monitor the translation of any specific protein, we select the “signature pair” of that protein, selected by our tRNA bioinformatics.

    The process is patent protected.

  • Target translation

    Target translation in Fibrosis:

    Target translation in Fibrosis: Steady state synthesis of collagen in fibroblast
    • Steady state synthesis of collagen in fibroblast
    • PSM: Lights OFF
    Target translation in Fibrosis: Activation of collagen synthesis in fibroblast
    • Activation of collagen synthesis in fibroblast
    • PSM: Lights ON

    Using FRET signals from signature tRNA pairs of the target protein, one tRNA of the pair is labeled with an energy donor, the other with an energy acceptor; we visualize and monitor the target’s translation.

  • Global translation

    Neuronal cells with transfected tRNA

    Anima’s PSM technology can monitor global translation within cells, providing spatial and temporal resolution. Translation of cell surface receptors is monitored on the Endoplasmic Reticulum (ER) or in the cytoplasm; in neurons translation is monitored at the cell body, in neurites or in axons.

    In addition to translation, tRNA transport along dendrites and axons may be monitored. Spatial resolution allows us to study the local translation response to signals at the synapse.

    Image: Neuronal cells with transfected tRNA

  • Pathway translation

    Single proteins or proteins belonging to the same family, or process, can be monitored by using a specific pair or pairs of tRNA. Thus, coordinated translation of proteins in response to external signals can be studied. The response of translation to regulators found upstream to biological process can be studied, for example inhibitors of mTOR or MAPK.

  • Target Identification

    Target Identification

    Post-transcription to translation regulatory mechanisms are novel targets that can be targeted with our technology. These include among others proteins that interact with the mRNA and its RBP’s (RNA Binding Proteins) and proteins that specifically regulate ribosome activity.

    Our platform discovers molecules that target these regulatory proteins and pathways. PSM not only identifies drug candidates but intrinsically helps in elucidating the pathways and mechanism of action of these molecules.

Proprietary Analysis

  • tRNA Bioinformatics

    tRNA Bioinformatics

    Our tRNAome bioinformatics identifies a unique tRNA pair that repeats at high frequency in the target protein’s sequence but is not frequently used in the background human proteome. Amazingly, 83% of known proteins have a tRNA pair with a repeat factor of 30 or more against the average background. When the selected “signature” pair is used in the screening system, a light pulse (FRET) is generated whenever the pair sits in a ribosome. This will happen at high frequency for the target but not for other proteins, enabling us to visually see the target protein’s translation. We can see when, where and how much of the target protein is being made in the cell, in real time.

    To monitor overall protein translation, all 48 types of labeled human tRNAs are used in the screen. This generates a light signal on each and every pair, creating a fascinating light map of the overall protein synthesis in the cells. The combined use of total tRNA and specific pair of tRNA enables differentiating global translation inhibitors and specific ones. Only compounds that show activity in inhibiting the translation with the target’s specific tRNA-pair but do not show activity on global translation (measured with total-tRNA) will be selected for further analysis.

  • Translation image analysis

    Translation image analysis

    Our proprietary image analysis extracts a large set of features from the millions of images of protein translation. Each feature represents a meaningful biological information item that is used in the generation of a translation signature profile.

    We use a data driven approach with proprietary data analysis methods that have been custom built for analyzing protein synthesis images:

    • The most informative features that reflect specific biological activity are selected
    • Features are independently selected computationally and cross-validated biologically
    • Compounds with known mechanism of action are tested, based on a test plate with known compounds
    • The biological and pharmacological relevance of the data is ensured by our data analysts working hand in hand with the biologists
    • "translation signature profiles" are created as a phenotype
    • cFRET features show high scores & reproducibility across different screens
    • Features like number of spots, brightness, localization and size are used as good indicators of a compound’s effect on the protein synthesis process
  • Big data algorithms

    We developed proprietary algorithms to analyze the huge amounts of data in dozens of millions of protein synthesis images that are generated in a typical screen. These include:

    • Classifications
    • Machine learning
    • Dynamic thresholding
    • Designated algorithmic approach / project : for each screen, some new elements in the analysis are specifics for cell-lines and indication

    In the image: Each cell population define a volume in space. This multi-dimensional approach allows us to point out possible hits regardless of their distribution.

  • Hit selection, validation and de-risking

    Analysis is done in several iterations by applying multiple secondary assays built around our technology. We analyze the impact of hit compounds on overall protein synthesis, eliminating compounds that are toxic, that interfere with the synthesis of many of the cell's proteins or that shut down the translation machinery itself. Additional translation selectivity assays were developed which monitor the effect of compounds on groups of proteins of interest or specific proteins along a pathway.

    PSM (protein synthesis monitoring) ASSAYS
    PSM analysis of compounds in target cells (at least two different cells)
    PSM analysis using cell free translation systems
    PSM signature of safe drugs

    NON PSM-ASSAYS:
    Effect on mRNA levels and localization
    Monitoring general protein synthesis: newly synthesized proteins
    Monitoring translation regulatory proteins
    Ribosome footprint coupled to proteomics

    Compound safety: Library of approved drugs is screened against PSM translation panel
    Comparing the compound’s translation impact profile against the profiles of known, safe drugs
    Di-tRNA combinations representing protein families
    Several cell types: epithelial (kidney, liver) and fibroblasts

Cloud Architecture

  • Anima's cloud-based system is hosted on Amazon (AWS) in a private secured zone, using the highest security, access control and data recovery standards.

    The platform implements an end to end, fully automated screening and analysis process: Plate data from screening machines is uploaded to the cloud in real time. Proprietary big data analysis algorithms analyze millions of images and billions of data points to identify compounds that are active as translation modulators of the target protein.

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